John N Galgiani
- Director, Valley Fever Center for Excellence
- Professor, Medicine
- Professor, BIO5 Institute
- Member of the Graduate Faculty
- Professor, Immunobiology
- Professor, Internal Medicine
Dr. Galgiani has focused his career on Arizona’s special problems with Valley Fever. His work has included studies of the impact of Valley Fever on the general population and on special groups such as organ transplant recipients and patients with AIDS. For 19 years, as part of the NIH-sponsored Mycoses Study Group, Dr. Galgiani has been the project director of a coccidioidomycosis clinical trials group. Through collaboration, this group has evaluated new therapies for Valley Fever more rapidly and with greater clarity than might otherwise have been possible by investigators working in isolation. Dr. Galgiani has also been involved with efforts to prevent Valley Fever through vaccination. His group discovered and patented a recombinant antigen which is the basis for a vaccine candidate suitable for further development and clinical trials. He is currently leading a program to develop a live, avirulent vaccine candidate discovered by others at the UA. He is also the project leader for developing a new drug, nikkomycin Z, for treating Valley Fever. With NIH and FDA grant awards, clinical trials with this drug were resumed in 2007. Dr. Galgiani is also Chief Medical Officer of Valley Fever Solutions, Inc, a start-up company founded to assist in the drug’s development.
In 1996, the Arizona Board of Regents accepted Dr. Galgiani’s proposal to establish the Valley Fever Center for Excellence for the Arizona universities. Based at the University of Arizona, the Center is pledged to spread information about Valley Fever, help patients with the severest complications of this disease, and to encourage research into the biology and diseases of its etiologic agent. The Center maintains a website (www.VFCE.Arizona.edu) and answers inquiries from health care professionals located in Arizona, other parts of the United States, and even from other countries. The Valley Fever Corridor Project, begun in 2009, intends to facilitate communication among Arizona clinicians to also improve patient care. Research is increasing into the environmental biology of the fungus within its desert soil habitat as well as how the fungus caused disease and the body’s immunity controls it. Since Arizona has the only medical schools situated directly within the endemic region for Valley Fever, it is quite appropriate that Arizona lead in solving this problem. As Director of the Center, Dr. Galgiani is working for its full implementation as a means of ensuring an institutional commitment to accomplish this goal.
- M.D. Medicine
- Northwestern Medical School, Chicago, Illinois, United States
- B.S. Biology
- Stanford University, Palo Alto, California, United States
- Valley Fever Solutions, Inc (2007 - Ongoing)
- University of Arizona, Tucson, Arizona (1996 - Ongoing)
- University of Arizona, Tucson, Arizona (1990 - Ongoing)
- University of Arizona, Tucson, Arizona (1984 - 2009)
- Southern Arizona VA Health Care System (1978 - 2009)
- University of Arizona, Tucson, Arizona (1978 - 1984)
- Stanford University, Palo Alto, California (1974 - 1976)
- Infectious Diseases Society of America, Spring 1987
- American College of Physicians, Spring 1982
- Distinguished Chair/Director
- University of Arizona, Spring 2022
- Sigma XI Scientific Research Honor Society, Fall 2021
- Clinical Researcher of the Year, finalist
- Tucson Business 2019 Influential Health and Medical Leaders, Summer 2019 (Award Finalist)
- Public Health Service Award
- Arizona Medical Society, Summer 2012
- Arizona Medical Association, Spring 2012
- Innovation Day Leading Edge Researcher
- University of Arizona, Fall 2008
- Founders Day Award
- UA COM-Tucson, Fall 2003
- University of Arizona College of Medicine, Fall 2003
- Achievement Award
- Coccidioidomycosis Study Group, Spring 2001
Licensure & Certification
- Medical Licensure, California, Medical Board of California (1973)
- Board Certification, American Board of Internal Medicine (1978)
- Medical Licensure, Arizona, Arizona Medical Board (1978)
Internship in Applied BiosciABS 593A (Spring 2023)
Internship in Applied BiosciABS 593A (Fall 2022)
Honors ThesisACBS 498H (Spring 2018)
Honors ThesisACBS 498H (Fall 2017)
- Brown, H. E., Comrie, A. C., Tamerius, J., Khan, M., Tabor, J. A., & Galgiani, J. N. (2014). Climate, wind storms, and the risk of valley fever (coccidioidomycosis). In The Influence of Global Environmental Change on Infectious Disease Dynamics(pp Chapter A12). Washington, D.C.: The National Academies Press.
- Donovan, F. M., Ramadan, F. A., Lim, J. R., Buchfuhrer, J. E., Khan, R. N., DeQuillfeldt, N. P., Davis, N. M., Kaveti, A., De Shadarevian, M., Bedrick, E. J., & Galgiani, J. N. (2022). Contribution of Biologic Response Modifiers to the Risk of Coccidioidomycosis Severity. Open forum infectious diseases, 9(3), ofac032.More infoThe risk of coccidioidomycosis (CM) as a life-threatening respiratory illness or disseminated CM (DCM) increases as much as 150-fold in immunosuppressed patients. The safety of biologic response modifiers (BRMs) as treatment for patients with autoimmune disease (AI) in CM-endemic regions is not well defined. We sought to determine that risk in the Tucson and Phoenix areas.
- Galgiani, J. N., Pu, J., Miranda, V., Minior, D., Raynolds, S., & Rayhorn, B. (2022). 470. Changing Urgent Care Patterns of Diagnosing Coccidioidomycosis in a Highly Endemic Urban Population. Open Forum Infectious Diseases, 9(Supplement_2). doi:10.1093/ofid/ofac492.528
- Galgiani, J. N., Shubitz, L. F., Orbach, M. J., Mandel, M. A., Powell, D. A., Klein, B. S., Robb, E. J., Ohkura, M., Seka, D. J., Tomasiak, T. M., & Monath, T. P. (2022). Vaccines to Prevent Coccidioidomycosis: A Gene-Deletion Mutant of Coccidioides Posadasii as a Viable Candidate for Human Trials. Journal of fungi (Basel, Switzerland), 8(8).More infoCoccidioidomycosis is an endemic fungal infection that is reported in up to 20,000 persons per year and has an economic impact close to $1.5 billion. Natural infection virtually always confers protection from future exposure, and this suggests that a preventative vaccine strategy is likely to succeed. We here review progress toward that objective. There has been ongoing research to discover a coccidioidal vaccine over the past seven decades, including one phase III clinical trial, but for reasons of either efficacy or feasibility, a safe and effective vaccine has not yet been developed. This review first summarizes the past research to develop a coccidioidal vaccine. It then details the evidence that supports a live, gene-deletion vaccine candidate as suitable for further development as both a veterinary and a human clinical product. Finally, a plausible vaccine development plan is described which would be applicable to this vaccine candidate and also useful to other future candidates. The public health and economic impact of coccidioidomycosis fully justifies a public private partnership for vaccine development, and the development of a vaccine for this orphan disease will likely require some degree of public funding.
- Hsu, A. P., Korzeniowska, A., Aguilar, C. C., Gu, J., Karlins, E., Oler, A. J., Chen, G., Reynoso, G. V., Davis, J., Chaput, A., Peng, T., Sun, L., Lack, J. B., Bays, D. J., Stewart, E. R., Waldman, S. E., Powell, D. A., Donovan, F. M., Desai, J. V., , Pouladi, N., et al. (2022). Immunogenetics associated with severe coccidioidomycosis. JCI insight, 7(22).More infoDisseminated coccidioidomycosis (DCM) is caused by Coccidioides, pathogenic fungi endemic to the southwestern United States and Mexico. Illness occurs in approximately 30% of those infected, less than 1% of whom develop disseminated disease. To address why some individuals allow dissemination, we enrolled patients with DCM and performed whole-exome sequencing. In an exploratory set of 67 patients with DCM, 2 had haploinsufficient STAT3 mutations, and defects in β-glucan sensing and response were seen in 34 of 67 cases. Damaging CLEC7A and PLCG2 variants were associated with impaired production of β-glucan-stimulated TNF-α from PBMCs compared with healthy controls. Using ancestry-matched controls, damaging CLEC7A and PLCG2 variants were overrepresented in DCM, including CLEC7A Y238* and PLCG2 R268W. A validation cohort of 111 patients with DCM confirmed the PLCG2 R268W, CLEC7A I223S, and CLEC7A Y238* variants. Stimulation with a DECTIN-1 agonist induced DUOX1/DUOXA1-derived hydrogen peroxide [H2O2] in transfected cells. Heterozygous DUOX1 or DUOXA1 variants that impaired H2O2 production were overrepresented in discovery and validation cohorts. Patients with DCM have impaired β-glucan sensing or response affecting TNF-α and H2O2 production. Impaired Coccidioides recognition and decreased cellular response are associated with disseminated coccidioidomycosis.
- Mandel, M. A., Beyhan, S., Voorhies, M., Shubitz, L. F., Galgiani, J. N., Orbach, M. J., & Sil, A. (2022). The WOPR family protein Ryp1 is a key regulator of gene expression, development, and virulence in the thermally dimorphic fungal pathogen Coccidioides posadasii. PLoS pathogens, 18(4), e1009832.More infoCoccidioides spp. are mammalian fungal pathogens endemic to the Southwestern US and other desert regions of Mexico, Central and South America, with the bulk of US infections occurring in California and Arizona. In the soil, Coccidioides grows in a hyphal form that differentiates into 3-5 micron asexual spores (arthroconidia). When arthroconidia are inhaled by mammals they undergo a unique developmental transition from polar hyphal growth to isotropic expansion with multiple rounds of nuclear division, prior to segmentation, forming large spherules filled with endospores. Very little is understood about the molecular basis of spherule formation. Here we characterize the role of the conserved transcription factor Ryp1 in Coccidioides development. We show that Coccidioides Δryp1 mutants have altered colony morphology under hypha-promoting conditions and are unable to form mature spherules under spherule-promoting conditions. We analyze the transcriptional profile of wild-type and Δryp1 mutant cells under hypha- and spherule-promoting conditions, thereby defining a set of hypha- or spherule-enriched transcripts ("morphology-regulated" genes) that are dependent on Ryp1 for their expression. Forty percent of morphology-regulated expression is Ryp1-dependent, indicating that Ryp1 plays a dual role in both hyphal and spherule development. Ryp1-dependent transcripts include key virulence factors such as SOWgp, which encodes the spherule outer wall glycoprotein. Concordant with its role in spherule development, we find that the Δryp1 mutant is completely avirulent in the mouse model of coccidioidomycosis, indicating that Ryp1-dependent pathways are essential for the ability of Coccidioides to cause disease. Vaccination of C57BL/6 mice with live Δryp1 spores does not provide any protection from lethal C. posadasii intranasal infection, consistent with our findings that the Δryp1 mutant fails to make mature spherules and likely does not express key antigens required for effective vaccination. Taken together, this work identifies the first transcription factor that drives mature spherulation and virulence in Coccidioides.
- O'Shaughnessy, E., Yasinskaya, Y., Dixon, C., Higgins, K., Moore, J., Reynolds, K., Ampel, N. M., Angulo, D., Blair, J. E., Catanzaro, A., Galgiani, J. N., Garvey, E., Johnson, R., Larwood, D. J., Lewis, G., Purdie, R., Rex, J. H., Shubitz, L. F., Stevens, D. A., , Page, S. J., et al. (2022). FDA Public Workshop Summary-Coccidioidomycosis (Valley Fever): Considerations for Development of Antifungal Drugs. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 74(11), 2061-2066.More infoCoccidioidomycosis is a fungal disease endemic to the southwestern United States, Mexico, and Central and South America. Prevalence rates are increasing steadily, and new endemic areas of Coccidioides are emerging. Standard treatment is often administered for months to decades, and intolerance to medications and treatment failures are common. No new treatments for coccidioidomycosis have been approved in the United States in nearly 40 years. On 5 August 2020, the US Food and Drug Administration convened experts in coccidioidomycosis from academia, industry, patient groups, and other government agencies to discuss the disease landscape and strategies to facilitate product development for treatment of coccidioidomycosis. This article summarizes the key topics concerning drug development for coccidioidomycosis presented by speakers and panelists during the workshop, such as unmet need, trial designs, endpoints, incentives, research and development support, and collaborations to facilitate antifungal drug development.
- Powell, D. A., Hsu, A. P., Shubitz, L. F., Butkiewicz, C. D., Moale, H., Trinh, H. T., Doetschman, T., Georgieva, T. G., Reinartz, D. M., Wilson, J. E., Orbach, M. J., Holland, S. M., Galgiani, J. N., & Frelinger, J. A. (2022). Mouse Model of a Human STAT4 Point Mutation That Predisposes to Disseminated Coccidiomycosis. ImmunoHorizons, 6(2), 130-143.More infoSTAT4 plays a critical role in the generation of both innate and adaptive immune responses. In the absence of STAT4, Th1 responses, critical for resistance to fungal disease, do not occur. Infection with the dimorphic fungus, , is a major cause of community-acquired pneumonia in the endemic regions of Arizona and California. In some people and often for unknown reasons, coccidioidal infection results in hematogenous dissemination and progressive disease rather than the typical self-limited pneumonia. Members of three generations in a family developed disseminated coccidioidomycosis, prompting genetic investigation. All affected family members had a single heterozygous base change in , c.1877A>G, causing substitution of glycine for glutamate at AA626 ( ). A knockin mouse, heterozygous for the substitution, developed more severe experimental coccidioidomycosis than did wild-type mice. T cells were deficient in production of IFN-γ after anti-CD3/CD28 stimulation. Spleen cells from mice showed defective responses to IL-12/IL-18 stimulation in vitro. In vivo, early postinfection, mutant mice failed to produce IFN-γ and related cytokines in the lung and to accumulate activated adaptive immune cells in mediastinal lymph nodes. Therefore, defective early induction of IFN-γ and adaptive responses by STAT4 prevents normal control of coccidioidomycosis in both mice and humans.
- Ramadan, F. A., Ellingson, K. D., Canales, R. A., Bedrick, E. J., Galgiani, J. N., & Donovan, F. M. (2022). Cross-Sectional Study of Clinical Predictors of Coccidioidomycosis, Arizona, USA. Emerging infectious diseases, 28(6), 1091-1100.More infoDemographic and clinical indicators have been described to support identification of coccidioidomycosis; however, the interplay of these conditions has not been explored in a clinical setting. In 2019, we enrolled 392 participants in a cross-sectional study for suspected coccidioidomycosis in emergency departments and inpatient units in Coccidioides-endemic regions. We aimed to develop a predictive model among participants with suspected coccidioidomycosis. We applied a least absolute shrinkage and selection operator to specific coccidioidomycosis predictors and developed univariable and multivariable logistic regression models. Univariable models identified elevated eosinophil count as a statistically significant predictive feature of coccidioidomycosis in both inpatient and outpatient settings. Our multivariable outpatient model also identified rash (adjusted odds ratio 9.74 [95% CI 1.03-92.24]; p = 0.047) as a predictor. Our results suggest preliminary support for developing a coccidioidomycosis prediction model for use in clinical settings.
- Shemuel, J., Bays, D. J., Thompson, G. R., Reef, S., Snyder, L., Freifeld, A. J., Huppert, M., Salkin, D., Wilson, M. D., & Galgiani, J. N. (2022). Natural history of pulmonary coccidioidomycosis: Further examination of the VA-Armed Forces Database. Medical mycology, 60(10).More infoThere are still many limitations related to the understanding of the natural history of differing forms of coccidioidomycosis (CM), including characterizing the spectrum of pulmonary disease. The historical Veterans Administration-Armed Forces database, recorded primarily before the advent of antifungal therapy, presents an opportunity to characterize the natural history of pulmonary CM. We performed a retrospective cohort study of 342 armed forces service members who were diagnosed with pulmonary CM at VA facilities between 1955 to 1958, followed through 1966, who did not receive antifungal therapy. Patients were grouped by predominant pulmonary finding on chest radiographs. The all-cause mortality was low for all patients (4.6%). Cavities had a median size of 3-3.9 cm (IQR: 2-2.9-4-4.9 cm), with heterogeneous wall thickness and no fluid level, while nodules had a median size of 1-1.19 cm (Interquartile range [IQR] 1-1.9-2-2.9 cm) and sharp borders. The majority of cavities were chronic (85.6%), and just under half were found incidentally. Median complement fixation titers in both the nodular and cavitary groups were negative, with higher titers in the cavitary group overall. This retrospective cohort study of non-disseminated coccidioidomycosis, the largest to date, sheds light on the natural history, serologic markers, and radiologic characteristics of this understudied disease. These findings have implications for the evaluation and management of CM.
- Shubitz, L. F., Powell, D. A., Dial, S. M., Butkiewicz, C. D., Trinh, H. T., Hsu, A. P., Buntzman, A., Frelinger, J. A., & Galgiani, J. N. (2022). Reactivation of Coccidioidomycosis in a Mouse Model of Asymptomatic Controlled Disease. Journal of fungi (Basel, Switzerland), 8(10).More infoThe majority of human coccidioidomycosis infections are asymptomatic or self-limited but may have sequestered spherules in highly structured granulomas. Under immunosuppression, reactivation of fungal growth can result in severe disease. B6D2F1 mice asymptomatically infected with strain 1038 were immunosuppressed with dexamethasone (DXM) in drinking water. Treated mice died 16-25 days later, while untreated mice survived ( < 0.001). Flow cytometry of lung granulomas on days 5, 10, 15, and 20 of DXM treatment showed immune cell populations decreased 0.5-1 log compared with untreated mice though neutrophils and CD19IgDIgM cells rebounded by day 20. Histopathology demonstrated loss of granuloma structure by day 5 and increasing spherules through day 20. On day 20, T-cells were nearly absent and disorganized pyogranulomatous lesions included sheets of plasma cells and innumerable spherules. Mice given DXM for 14 days then stopped (DXM stop) survived 6 weeks (9/10). Lung fungal burdens were significantly lower ( = 0.0447) than mice that continued treatment (DXM cont) but higher than untreated mice. Histopathologically, DXM stop mice did not redevelop controlled granulomas by sacrifice, though T-cells were densely scattered throughout the lesions. This demonstrates a mouse model suitable for further study to understand the immunologic components responsible for maintenance control of coccidioidomycosis.
- Thompson, G. R., Ampel, N. M., Blair, J. E., Donovan, F., Fierer, J., Galgiani, J. N., Heidari, A., Johnson, R., Shatsky, S. A., Uchiyama, C. M., & Stevens, D. A. (2022). Controversies in the Management of Central Nervous System Coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 75(4), 555-559.More infoCentral nervous system infection with Coccidioides spp. is fatal if untreated and complications occur even when therapy is directed by experienced clinicians. We convened a panel of clinicians experienced in the management of coccidioidal meningitis to summarize current controversies and provide consensus for the management of this difficult infection.
- Grizzle, A. J., Wilson, L., Nix, D. E., & Galgiani, J. N. (2021). Clinical and Economic Burden of Valley Fever in Arizona: An Incidence-Based Cost-of-Illness Analysis. Open forum infectious diseases, 8(2), ofaa623.More infoCoccidioidomycosis, ie, Valley fever, is an important fungal infection in the Southwest, with half to two thirds of all cases occurring in Arizona. This endemic respiratory disease can range from primary uncomplicated pneumonia to disseminated infection such as meningitis with chronic pulmonary complications. Valley fever diagnoses have risen over recent years and cause substantial morbidity and economic burden in Arizona.
- Menghani, S. V., Rivera, A., Neubert, M., Hagerty, J. R., Lewis, L., Galgiani, J. N., Jolly, E. R., Alvin, J. W., & Johnson, M. D. (2021). Demonstration of ,-Dimethyldithiocarbamate as a Copper-Dependent Antibiotic against Multiple Upper Respiratory Tract Pathogens. Microbiology spectrum, 9(2), e0077821.More infoTransition metals are necessary cofactors and structural elements in living systems. Exposure to high concentrations of biologically important transition metals, such as zinc and copper, results in cell toxicity. At the infection site, the immune system deploys metal sorbent proteins (e.g., lactoferrin and calprotectin) to starve pathogens of necessary metals (such as iron), while phagocytes expose engulfed pathogens to high levels of other metals, such as copper and zinc. The opportunistic pathogen Streptococcus pneumoniae (the pneumococcus) encounters macrophages during initial and protracted infections. The pneumococcus employs a copper export pathway, which improves colonization and persistent infection of the nasopharynx and the upper respiratory tract. Because copper is tightly regulated in the host, we instead sought to leverage the localized power of nutritional immunity by identifying small molecules with copper-dependent toxicity (CDT) through a targeted screen of compounds for antibiotic efficacy. We chose to include dithiocarbamates, based on the copper synergy observed in other organisms with 1-(diethylthiocarbamoyldisulfanyl)-,-diethyl-methanethioamide (tetraethylthiuram disulfide, disulfiram). We observed CDT of some dithiocarbamates in S. pneumoniae. Only ,-dimethyldithiocarbamate (DMDC) was consistently toxic across a range of concentrations with copper both and against the pneumococcus. We also observed various degrees of CDT using DMDC in Staphylococcus aureus, Coccidioides posadasii, and Schistosoma mansoni. Collectively, we demonstrate that the compound DMDC is a potent bactericidal compound against S. pneumoniae with antimicrobial efficacy against bacterial and fungal pathogens. With the rise of antibiotic resistance, approaches that add new antimicrobials to the current repertoire are vital. Here, we investigate putative and known copper ionophores in an attempt to intoxicate bacteria and use ionophore/copper synergy, and we ultimately find success with ,-dimethyldithiocarbamate (DMDC). We show that DMDC has efficacy in a copper-dependent manner and kills pathogens across three different kingdoms, Streptococcus pneumoniae, , and Schistosoma mansoni, and efficacy against S. pneumoniae. As such, dithiocarbamates represent a new potential class of antimicrobials and thus warrant further mechanistic investigation.
- Messina, J. A., Maziarz, E. K., Galgiani, J., Truong, J. T., Htoo, A. K., Heidari, A., Johnson, R. H., Narang, A. T., Donovan, F. M., Ewell, M., Catanzaro, A., Thompson, G. R., Ampel, N. M., Perfect, J. R., Naggie, S., & Walter, E. B. (2021). A randomized, double-blind, placebo-controlled clinical trial of fluconazole as early empiric treatment of coccidioidomycosis pneumonia (Valley Fever) in adults presenting with community-acquired pneumonia in endemic areas (FLEET-Valley Fever). Contemporary clinical trials communications, 24, 100851.More infoCoccidioidomycosis is a fungal infection endemic in the southwestern United States (US). Primary pulmonary coccidioidomycosis (PPC) is a leading cause of community-acquired pneumonia (CAP) in this region, although its diagnosis is often delayed, leading to lag in antifungal treatment and subsequent morbidity. The impact of early empiric antifungal therapy as part of treatment for CAP in endemic areas on clinical outcomes is unknown.
- Peng, T., Zong, Y., Johnson, M. D., Menghani, S. V., Lewis, M. L., & Galgiani, J. N. (2021). A quantitative enzyme-linked immunoassay (ELISA) to approximate complement-fixing antibody titers in serum from patients with coccidioidomycosis. Diagnostic microbiology and infectious disease, 99(1), 115198.More infoCoccidioidomycosis is most frequently diagnosed serologically, and the quantitative test for complement-fixing antibodies is considered prognostically useful. Because complement-fixing antibody testing is complex, labor-intensive, and poorly standardized, an enzyme-linked immunoassay (ELISA) alternative would be attractive. In this report, we restrict the complement-fixing, antibody-binding domain to a 200-amino-acid recombinant peptide of the known antigen. Over-lapping truncations of this peptide do not bind complement-fixing antibodies, suggesting that the responsible epitope(s) are conformational. Further, anchoring the antigenic peptide to the ELISA plate by means of a C-terminal biotin-mimic peptide tag instead of allowing the peptide to randomly adhere to the plastic plate improves sensitivity of antibody detection by 1-2 logs in different sera. The newly developed ELISA shows a significant quantitative correlation with complement-fixing antibody titers. This ELISA shows potential as the basis for a new quantitative assay for coccidioidal antibodies.
- Powell, D. A., Hsu, A. P., Butkiewicz, C. D., Trinh, H. T., Frelinger, J. A., Holland, S. M., Galgiani, J. N., & Shubitz, L. F. (2021). Vaccine Protection of Mice With Primary Immunodeficiencies Against Disseminated Coccidioidomycosis. Frontiers in cellular and infection microbiology, 11, 790488.More infoDisseminated coccidioidomycosis (DCM), often a severe and refractory disease leading to poor outcomes, is a risk for people with certain primary immunodeficiencies (PID). Several DCM-associated PID (STAT4, STAT3, IFNγ, and Dectin-1) are modeled in mice. To determine if vaccination could provide these mice protection, mice with mutations in , , , (Dectin-1), and Rag-1 (T- and B-cell deficient) knockout (KO) mice were vaccinated with the live, avirulent, Δ vaccine strain and subsequently challenged intranasally with pathogenic Silveira strain. Two weeks post-infection, vaccinated mice of all strains except Rag-1 KO had significantly reduced lung and spleen fungal burdens (p
- Powell, D. A., Shubitz, L. F., Butkiewicz, C. D., Trinh, H. T., Donovan, F. M., Frelinger, J. A., & Galgiani, J. N. (2021). TNFα Blockade Inhibits Both Initial and Continued Control of Pulmonary. Frontiers in cellular and infection microbiology, 11, 796114.More infoTumor necrosis factor alpha (TNFα) is a pluripotent cytokine that is important in many infections, though its role in infection remains poorly understood. The need to understand TNFα in infection has increased recently with the widespread use of TNFα inhibitors for a wide variety of autoimmune conditions. Here, we couple the newly developed infection model using strain Cp1038 and C57BL/6 × DBA/2J F1 (B6D2F1) mice. B6D2F1 mice develop long-lasting control of Cp1038. Treatment of B6D2F1 mice with anti-TNFα antibodies permits significant fungal proliferation and death. Additionally, we show that antibody treatment limited to the first 2 weeks of infection was sufficient to induce this same loss of fungal control. Importantly, anti-TNFα antibody treatment initiated after fungal control leads to a loss of host control. These results highlight the importance of TNFα in both the initial control of murine and ongoing suppression of the fungal disease.
- Pu, J., Donovan, F. M., Ellingson, K., Leroy, G., Stone, J., Bedrick, E., & Galgiani, J. N. (2021). Clinician Practice Patterns That Result in the Diagnosis of Coccidioidomycosis Before or During Hospitalization. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 73(7), e1587-e1593.More infoCoccidioidomycosis (CM) is common and important within endemic regions, requiring specific testing for diagnosis. Long delays in diagnosis have been ascribed to ambulatory clinicians. However, how their testing practices have impacted patient care has not been systematically unexplored.
- Shubitz, L. F., Powell, D. A., Butkiewicz, C. D., Lewis, M. L., Trinh, H. T., Frelinger, J. A., Orbach, M. J., & Galgiani, J. N. (2021). A Chronic Murine Disease Model of Coccidioidomycosis Using Coccidioides posadasii, Strain 1038. The Journal of infectious diseases, 223(1), 166-173.More infoMurine infections with most Coccidioides spp. strains are lethal by 3 weeks, limiting the study of immune responses. Coccidioides posadasii, strain 1038 (Cp1038), while slowly lethal, resulted in protracted survival of C57BL/6 (B6) mice. In resistant (B6D2)F1/J mice, lung fungal burdens stabilized by week 4 without progression through week 16, better modeling human coccidioidal infections after their immunologic control. Immunodeficient tumor necrosis factor (Tnf) α knockout (KO) and interferon (Ifn) γ receptor 1 (Ifn-γr1) KO mice survived a median of 22.5 and 34 days, compared with 70 days in B6 mice (P = .001 and P < .01, respectively), though 14-day lung fungal burden studies showed little difference between Ifn-γr1 KO and B6 mice. B6 mice showed peak concentrations of key inflammatory lung cytokines, including interleukin 6, 23, and 17A, Tnf-α, and Ifn-γ, only after 4 weeks of infection. The slower progression in B6 and the acquired fungal burden stability in B6D2 mice after Cp1038 infection greatly increases the array of possible immunologic studies.
- Shubitz, L. F., Robb, E. J., Powell, D. A., Bowen, R. A., Bosco-Lauth, A., Hartwig, A., Porter, S. M., Trinh, H., Moale, H., Bielefeldt-Ohmann, H., Hoskinson, J., Orbach, M. J., Frelinger, J. A., & Galgiani, J. N. (2021). Δcps1 vaccine protects dogs against experimentally induced coccidioidomycosis. Vaccine, 39(47), 6894-6901.More infoCoccidioidomycosis is a significant health problem of dogs and humans in endemic regions, especially California and Arizona in the U.S. Both species would greatly benefit from a vaccine to prevent this disease. A live avirulent vaccine candidate, Δcps1, was tested for tolerability and efficacy to prevent pulmonary coccidioidomycosis in a canine challenge model. Vaccine injection-site reactions were transient and there were no systemic effects observed. Six of seven vaccine sites tested and all draining lymph nodes were sterile post-vaccination. Following infection with Coccidioides posadasii, strain Silveira, arthroconidia into the lungs, dogs given primary and booster vaccinations had significantly reduced lung fungal burdens (P = 0.0003) and composite disease scores (P = 0.0002) compared to unvaccinated dogs. Dogs vaccinated once had fungal burdens intermediate between those given two doses or none, but disease scores were not significantly different from unvaccinated (P = 0.675). Δcps1 was well-tolerated in the dogs and it afforded a high level of protection when given as prime and boost. These results drive the Δcps1 vaccine toward a licensed veterinary vaccine and support continued development of this vaccine to prevent coccidioidomycosis in humans.
- Bays, D. J., Thompson, G. R., Reef, S., Snyder, L., Freifeld, A. J., Huppert, M., Salkin, D., Wilson, M. D., & Galgiani, J. N. (2020). Natural History of Disseminated Coccidioidomycosis: Examination of the VA-Armed Forces Database. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.More infoThe natural history of non-central nervous system (CNS) disseminated coccidioidomycosis (DCM) has not been previously characterized. The historical VA-Armed Forces coccidioidomycosis patient group provides a unique cohort of patients not treated with standard antifungal therapy allowing for characterization of the natural history of coccidioidomycosis.
- Donovan, F. M., Ramadan, F. A., Khan, S. A., Bhaskara, A., Lainhart, W. D., Narang, A. T., Mosier, J. M., Ellingson, K. D., Bedrick, E. J., Saubolle, M. A., & Galgiani, J. N. (2020). Comparison of a Novel Rapid Lateral Flow Assay to Enzyme Immunoassay Results for Early Diagnosis of Coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.More infoCoccidioidomycosis (CM) is a common cause of community acquired pneumonia (CAP) where CM is endemic. Manifestations include self-limited pulmonary infection, chronic fibrocavitary pulmonary disease, and disseminated coccidioidomycosis (DCM). Most infections are identified by serological assays including enzyme-linked immunoassay (EIA), complement fixation (CF) and immunodiffusion (IMDF). These are time-consuming and take days to result, impeding early diagnosis. A new lateral flow assay (LFA, Sōna, IMMY, Norman OK) improves time-to-result to one hour.
- Fierer, J., Galgiani, J. N., Powell, D. A., Pouladi, N., Lussier, Y. A., Lionakis, M. S., Hsu, A. P., Holland, S. M., Galgiani, J. N., Frelinger, J. A., Davis, J., & Chaput, A. L. (2020). 42. Common Population Variants Cause Susceptibility to Disseminated Coccidioidomycosis. Open Forum Infectious Diseases, 7(Supplement_1), S22-S23. doi:10.1093/ofid/ofaa417.041More infoAbstract Background Coccidioides are endemic, dimorphic fungi found in soils of southwestern United States, Mexico and Central America. Infection occurs via inhalation of arthroconidia which swell, differentiate into spherules and rupture releasing endospores. While the majority of infected individuals will never report illness, roughly 1/3 seek medical attention for fungal pneumonia and ~1% of those present with disseminated coccidioidomycosis (DCM). IL12-IFNγ pathway mutations have been reported in DCM but are exceedingly rare and cannot account for the ~500–600 cases of DCM/year. Methods We performed whole exome sequencing on 66 individuals with DCM, retaining variants predicted damaging (CADD >15) with a population frequency < 10%. Results Homozygous CLEC7A c.714T >G; p.Y238* causing a truncated Dectin-1 receptor was overrepresented (OR=9.8449, 95% CI 3.0841 to 31.4260, P=0.0001). Dectin-1 signaling pathway variants included 3 homozygous and 11 heterozygous CLEC7A p.Y238* individuals, one each CLEC7A p.I223S and MALT1 p.R149Q and five PLCG2 p.R268W. Since Dectin-1 is the receptor for b-glucan, a major Coccidioides cell-wall component, we hypothesized that Dectin-1 pathway variants could affect fungal recognition and cellular response. Healthy control PBMCs stimulated with purified β-glucan or heat-killed Candida albicans induced 6-fold more TNFα than patients with homozygous or heterozygous CLEC7A, PLCG2 or MALT1 variants (P=0.0022, Ordinary one-way ANOVA). Additionally, one patient with a family history of DCM but lacking a defined mutation also failed to up-regulate TNFα after stimulation. Normalized TNF production from healthy control and DCM patient’s peripheral blood mononuclear cells Conclusion These data are consonant with increased dissemination in Clec7a-/- mice as well as in patients receiving anti-TNF biologics. These gene variants accounted for 31% of our DCM cohort (21/66 patients). This is the first demonstration of variants outside the IL12-IFNg pathway impairing fungal recognition and cellular response in coccidioidomycosis. Common heterozygous variants may be sufficient for disease susceptibility to highly pathogenic organisms. Disclosures Michail Lionakis, MD, ScD, Matinas BioPharma (Research Grant or Support)
- Galgiani, J. N., Grizzle, A. J., Nix, D. E., & Wilson, L. (2020). Clinical and Economic Burden of Valley Fever in Arizona: An Incidence-Based Cost-of-Illness Analysis. Open Forum On Infectious Diseases, 1-32. doi:https://doi.org/10.1093/ofid/ofaa623
- Galgiani, J. N., Orbach, M. J., Frelinger, J. A., Shubitz, L. F., Powell, D. A., Butkiewicz, C. D., Lewis, M. L., & Trinh, H. T. (2020). A Chronic Murine Disease Model of Coccidioidomycosis Using Coccidioides posadasii, Strain 1038. The Journal of Infectious Diseases, 223(1), 166-173. doi:10.1093/infdis/jiaa419
- Mead, H. L., Hamm, P. S., Shaffer, I. N., Teixeira, M. M., Wendel, C. S., Wiederhold, N. P., Thompson, G. R., Muñiz-Salazar, R., Castañón-Olivares, L. R., Keim, P., Plude, C., Terriquez, J., Galgiani, J. N., Orbach, M. J., & Barker, B. M. (2020). Differential Thermotolerance Adaptation between Species of. Journal of fungi (Basel, Switzerland), 6(4).More infoCoccidioidomycosis, or Valley fever, is caused by two species of dimorphic fungi. Based on molecular phylogenetic evidence, the genus contains two reciprocally monophyletic species: and . However, phenotypic variation between species has not been deeply investigated. We therefore explored differences in growth rate under various conditions. A collection of 39 C. posadasii and 46 C. immitis isolates, representing the full geographical range of the two species, was screened for mycelial growth rate at 37 °C and 28 °C on solid media. The radial growth rate was measured for 16 days on yeast extract agar. A linear mixed effect model was used to compare the growth rate of and at 37 °C and 28 °C, respectively. grew significantly faster at 37 °C, when compared to ; whereas both species had similar growth rates at 28 °C. These results indicate thermotolerance differs between these two species. As the ecological niche has not been well-described for spp., and disease variability between species has not been shown, the evolutionary pressure underlying the adaptation is unclear. However, this research reveals the first significant phenotypic difference between the two species that directly applies to ecological research.
- Berghout, J., Galgiani, J. N., Shubitz, L. F., Powell, D. A., Lussier, Y. A., Hsu, A. P., Holland, S. M., Galgiani, J. N., Frelinger, J. A., Davis, J., & Berghout, J. (2019). 2888. STAT4 Mutation in Three Generations with Disseminated Coccidioidomycosis (DCM) also Exhibits Increased Susceptibility to Coccidioidal Infection in Transfected Mice. Open Forum Infectious Diseases, 6(Supplement_2), S77-S78. doi:10.1093/ofid/ofz359.166More infoAbstract Background Reported coccidioidomycosis has increased with case rates of 198/100,000 in Arizona (2012). In California alone, 2000–2011 hospitalizations were $2.2B. Dissemination occurs in 8% of reports with significant morbidity and occasional deaths. DCM was found in 3 generations: grandmother (skin), mother (skin), and son (bone). Whole exome sequencing identified a heterozygous (het) STAT4 mutation (p.E626G) in all three. This mutation alters the phosphotyrosine binding pocket and is predicted to impair STAT4 function, interfering with (i) receptor binding and phosphorylation, (ii) nuclear localization, and/or (iii) transcription. Expression profiling of antigen-stimulated peripheral blood mononuclear cells from one patient showed dampening of known STAT4 targets compared with controls. Methods STAT4 p.E626G was generated and confirmed in C57BL/6NJ (WT) mice using CRISPR-Cas9. With continued breeding, neither homozygous (hom) nor het mice had gross abnormalities. There were normal spleen and lung lymphoid cell numbers. Thymus and bone marrow had normal development of lymphoid subsets. We performed intranasal infection with reduced virulence C. posadasii strain 1038 or with F. tularensis live vaccine- strain (LVS). Naïve or Δcps1-vaccinated mice were tested for resistance to C. posadasii strain Silveira. Results At day 21 post Cp 1038 infection, hom, het, and WT mice had similar lung fungal burdens (~104.7 cfu). All p.E626G mice died between days 31 and 39 with lung burden significantly higher (~9 × 106 cfu) than WT sacrificed on day 44 (7 × 105 cfu, P = 0.015). After LVS infection, p.E626G mice had increased lung bacterial cfu and all had dissemination to the spleen compared with WT lung bacterial burden and no splenic dissemination. Immunized het and WT mice all had significantly reduced lung cfu 14 days following C. posadasii infection compared with unvaccinated WT mice. Conclusion The STAT4 p.E626G mutated mouse recapitulated patients’ increased susceptibility to coccidioidal infection. The decreased fungal burdens seen in Δcps1-vaccinated mice suggest that vaccination may be effective in those persons genetically susceptible to DCM. Given the increasing frequency and economic burdens of coccidioidomycosis, pursuit of vaccination strategies should continue. Disclosures All Authors: No reported Disclosures.
- Donovan, F. M., Shubitz, L., Powell, D., Orbach, M., Frelinger, J., & Galgiani, J. N. (2019). Early Events in Coccidioidomycosis. Clinical microbiology reviews, 33(1).More infoSUMMARYSince its description nearly 130 years ago, hundreds of studies have deepened our understanding of coccidioidomycosis, also known as valley fever (VF), and provided useful diagnostic tests and treatments for the disease caused by the dimorphic fungi spp. In general, most of the literature has addressed well-established infections and has described patients who have experienced major complications. In contrast, little attention has been given to the earliest consequences of the pathogen-host interaction and its implications for disease manifestation, progression, and resolution. The purpose of this review is to highlight published studies on early coccidioidomycosis, identify gaps in our knowledge, and suggest new or former research areas that might be or remain fertile ground for insight into the early stages of this invasive fungal disease.
- Donovan, F. M., Wightman, P., Zong, Y., Gabe, L., Majeed, A., Ynosencio, T., Bedrick, E. J., & Galgiani, J. N. (2019). Delays in Coccidioidomycosis Diagnosis and Associated Healthcare Utilization, Tucson, Arizona, USA. Emerging infectious diseases, 25(9), 1745-1747.More infoTucson, Arizona, USA, is a highly coccidioidomycosis-endemic area. We conducted a retrospective review of 815 patients in Tucson over 2.7 years. Of 276 patients with coccidioidomycosis, 246 had a delay in diagnosis; median delay was 23 days. Diagnosis delay was associated with coccidioidomycosis-related costs totaling $589,053 and included extensive antibacterial drug use.
- Galgiani, J. N., Blair, J. E., Ampel, N. M., & Thompson, G. R. (2019). Treatment for Early, Uncomplicated Coccidioidomycosis: What is Success?. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.More infoThe care of primary pulmonary coccidioidomycosis remains challenging. Such infections produce a variety of signs, symptoms, and serologic responses that cause morbidity in patients and concern in treating clinicians for the possibility of extrapulmonary dissemination. Illness may be due to ongoing fungal growth that produces acute inflammatory responses, resulting in tissue damage and necrosis, and for this, administering an antifungal drug may be of benefit. In contrast, convalescence may be prolonged by other immunologic reactions to infection, even after fungal replication has been arrested, and in those situations, antifungal therapy is unlikely to yield clinical improvement. In this presentation, we discuss what findings are clinical indicators of fungal growth and what other sequalae are not. Understanding these differences provides a rational management strategy for deciding when to continue, discontinue, or reinstitute antifungal treatments.
- Hartwig, A., Galgiani, J. N., Trinh, H. T., Shubitz, L. F., Robb, E. J., Powell, D. A., Lewis, M. L., Hartwig, A. E., Galgiani, J. N., Frelinger, J. A., Bowen, R., & Bosco-lauth, A. M. (2019). 1732. A Canine Target Species Challenge Model to Evaluate Efficacy of a Coccidioidomycosis Vaccine. Open Forum Infectious Diseases, 6(Supplement_2), S634-S635. doi:10.1093/ofid/ofz360.1595More infoAbstract Background The preferred efficacy design for licensing a vaccine for animal use (United States Department of Agriculture (USDA)) is a prospective, placebo-controlled, randomized, and double-blinded vaccination-challenge trial. In such studies, each subject receives the same exposure to the virulent pathogen by active challenge. To test a cps1, live avirulent canine coccidioidomycosis vaccine, an inhalation disease model was developed in beagle dogs. Methods 6-month old male beagle dogs were housed according to PHS standards. All procedures, approved by the Institutional Animal Care and Use Committee for Colorado State University, were performed at ABSL3. Dogs were infected by nebulization with low, medium or high counts of arthroconidia of Coccidioides posadasii, strain Silveira, delivered via endotracheal tube under injectable anesthesia. Thoracic radiographs, CBC, and serum chemistries and body weights were obtained at 2- or 3-week intervals and dogs were euthanized 8 weeks p.i., or earlier if necessary. Approximately 1 gram lung specimens from each lobe were cultured for fungal burden. Fixed tissues were examined histologically. Serum was tested for antibodies. Results Ten of 11 dogs were successfully infected; 5 required early removal at 33 to 48-days p.i. Elevated globulin, decreased albumin, decreased A/G ratio, monocytosis and weight loss were present in all infected dogs. Radiographic and histopathological lesions were very extensive at the high challenge doses. Medium doses had the most consistent scoring and clinical findings, including some early removal, without overwhelming disease, while the low dose produced the least consistent quantifiable features. All dogs developed antibodies. Conclusion Nebulized aerosol delivery of spores reproducibly produced significant coccidioidomycosis in 10 of 11 dogs. Overall, the challenge model demonstrated consistent characteristic findings sufficient to assess vaccine efficacy in dogs during an 8-week period post challenge without producing a potentially overwhelming infection. The aerosol nebulization of arthroconidia in beagle dogs should provide a vaccination-challenge experimental design in line with Chapter 9 Code of Federal Regulations, parts 102.5 and 104.5. Disclosures All authors: No reported disclosures.
- Odio, C. D., Leng, L., Siu, E., Piecychna, M., Galgiani, J. N., Holland, S. M., & Bucala, R. (2019). 2598. Macrophage Migration Inhibitory Factor May Contribute to Disseminated Coccidioidomycosis Susceptibility. Open Forum Infectious Diseases, 6(Supplement_2), S903-S903. doi:10.1093/ofid/ofz360.2276More infoAbstract Background Disseminated coccidioidomycosis occurs in
- Teixeira, M. M., Alvarado, P., Roe, C. C., Thompson, G. R., Patané, J. S., Sahl, J. W., Keim, P., Galgiani, J. N., Litvintseva, A. P., Matute, D. R., & Barker, B. M. (2019). Population Structure and Genetic Diversity among Isolates of in Venezuela and Surrounding Regions. mBio, 10(6).More infois a pathogenic fungus that causes coccidioidomycosis in many arid regions of the Americas. One of these regions is bordered by the Caribbean Sea, and the surrounding landscape may play an important role in the dispersion of across South America through southeastern Mexico, Honduras, Guatemala, and Venezuela. Comparative phylogenomic analyses of reveal that clinical strains from Venezuela are genetically distinct from the North American populations found in (i) Arizona and (ii) Texas, Mexico, and the rest of South America (TX/MX/SA). We find evidence for admixture between the Venezuela and the North American populations of in Central America. Additionally, the proportion of Venezuelan alleles in the admixed population decreases as latitude (and distance from Venezuela) increases. Our results indicate that the population in Venezuela may have been subjected to a recent bottleneck and shows a strong population structure. This analysis provides insight into potential for spp. to invade new regions. Valley Fever is a fungal disease caused by two species of fungi: and These fungi are found throughout the arid regions of North and South America; however, our understanding of genetic diversity and disease in South America is limited. In this report, we analyze 10 new genomes of from regions bordering the Caribbean Sea. We show that these populations are distinct and that isolates from Venezuela are likely a result of a recent bottleneck. These data point to patterns that might be observed when investigating recently established populations.
- Van Dyke, M. C., Thompson, G. R., Galgiani, J. N., & Barker, B. M. (2019). The Rise of : Forces Against the Dust Devil Unleashed. Frontiers in immunology, 10, 2188.More infoCoccidioidomycosis (Valley fever) is a fungal disease caused by the inhalation of or . This neglected disease occurs in the desert areas of the western United States, most notably in California and Arizona, where infections continue to rise. Clinically, coccidioidomycosis ranges from asymptomatic to severe pulmonary disease and can disseminate to the brain, skin, bones, and elsewhere. New estimates suggest as many as 350,000 new cases of coccidioidomycosis occur in the United States each year. Thus, there is an urgent need for the development of a vaccine and new therapeutic drugs against infection. In this review, we discuss the battle against including the development of potential vaccines, the quest for new therapeutic drugs, and our current understanding of the protective host immune response to infection.
- Hung, C. Y., Zhang, H., Castro-Lopez, N., Ostroff, G. R., Khoshlenar, P., Abraham, A., Cole, G. T., Negron, A., Forsthuber, T., Peng, T., Galgiani, J. N., Ampel, N. M., & Yu, J. J. (2018). Glucan-chitin particles enhance Th17 response and improve protective efficacy of a multivalent antigen (rCpa1) against pulmonary infection. Infection and immunity.More infoDeveloping an effective and safe recombinant vaccine requires microbe-specific antigens combined with an adjuvant/delivery system to strengthen protective immunity. In this study, we designed and expressed a multivalent, recombinant polypeptide antigen (rCpa1) that consists of three previously identified antigens (i.e., Ag2/Pra, Cs-Ag and Pmp1) and 5 pathogen-derived peptides with high affinity for human MHC II molecules. The purified rCpa1 was encapsulated into four types of yeast cell-wall particles containing various compositions of β-glucan, mannan and chitin or mixed with an oligonucleotide (ODN) containing 2 methylated dinucleotide CpG motifs. This multivalent antigen encapsulated into glucan-chitin particles (GCP-rCpa1) showed a significantly elevated reduction of fungal burden for human HLA-DR4 transgenic mice compared to the other tested adjuvant-rCpa1 formulations. Among the tested adjuvants GCPs and GPs were both capable of stimulating a mixed Th1 and Th17 response. Mice vaccinated with GCP-rCpa1 showed elevated IL-17 production in T-cell recall assays and early lung infiltration of activated Th1 and Th17 cells compared to GP-rCpa1-vaccinated mice. Both C57BL/6 and HLA-DR4 transgenic mice that were vaccinated with GCP-rCpa1 vaccine increased surivial compared to the mice received GCPs alone. Concurringly, GCP-rCpa1 vaccine stimulated enhanced infiltration of macrophages to engulf and process the vaccine for antigen presentation in the injection sites compared to GP-rCpa1 injection. This is the first attempt to systematically characterize the presentation of a multivalent coccidioidomycosis vaccine encapsulated with selected adjuvants which enhance protective cellular immune response to infection.
- Mead, H. L., Teixeira, M. M., Galgiani, J. N., & Barker, B. M. (2018). Characterizing in vitro spherule morphogenesis of multiple strains of both species of Coccidioides. Medical mycology.More infoThe disease San Joaquin Valley Fever (coccidioidomycosis) is caused by the inhalation of Coccidioides arthroconidia. In vivo, arthroconidia transform into pathogenic structures termed spherules. Exposure to the host milieu triggers spherule development; however, the molecular mechanisms responsible for the morphological shift are not well characterized. This study compared the morphogenesis of five strains of both species of Coccidioides in two media types to improve the in vitro model of dimorphism that can be easily reproduced, and is amenable to tissue culture. We also sought to establish a modern record of the morphological switch among commonly used lab strains through a detailed account of growth under various conditions. Spherules from five strains were grown in standard (Converse) and experimental media (RPMI-sph). Strain behavior was quantified by median spherule size and spherule concentration, beginning 3 days after inoculation and followed for 10 days of growth. There were significant differences observed among Coccidioides immitis and C. posadasii strains, as well as differences between the in vitro systems.
- Shubitz, L. F., Powell, D. A., Trinh, H. T., Lewis, M. L., Orbach, M. J., Frelinger, J. A., & Galgiani, J. N. (2018). Viable spores of Coccidioides posadasii Δcps1 are required for vaccination and provide long lasting immunity. Vaccine, 36(23), 3375-3380.More infoCoccidioidomycosis is a systemic fungal infection for which a vaccine has been sought for over fifty years. The avirulent Coccidioides posadasii strain, Δcps1, which is missing a 6 kb gene, showed significant protection in mice. These studies explore conditions of protection in mice and elucidate the immune response. Mice were vaccinated with different doses and viability states of Δcps1 spores, challenged with virulent C. posadasii, and sacrificed at various endpoints, dependent on experimental objectives. Tissues from vaccinated mice were harvested for in vitro elucidation of immune response. Vaccination with viable Δcps1 spores was required for protection from lethal challenge. Viable spore vaccination produced durable immunity, lasting at least 6 months, and prolonged survival (≥6 months). The C. posadasii vaccine strain also protected mice against C. immitis (survival ≥ 6 months). Cytokines from infected lungs of vaccinated mice in the first four days after Cp challenge showed significant increases of IFN-γ, as did stimulated CD4 spleen cells from vaccinated mice. Transfer of CD4 cells, but not CD8 or B cells, reduced fungal burdens following challenge. IFN-γ from CD4 cells in vaccinated mice indicates a Th1 response, which is critical for host control of coccidioidomycosis.
- Donovan, F. M., Zangeneh, T. T., Malo, J., & Galgiani, J. N. (2017). Top Questions in the Diagnosis and Treatment of Coccidioidomycosis. Open forum infectious diseases, 4(4), ofx197.More infoRevised and greatly expanded treatment guidelines for coccidioidomycosis were published last year by the Infectious Diseases Society of America. We have selected 4 questions that commonly arise in the management of patients suspected of this disease and for which there remain divided opinions.
- Donovan, F., Malo, J., Zangeneh, T. T., & Galgiani, J. N. (2017). Top Questions in Diagnosis and Treatment of Coccidioidomycosis. Open Forum Infectious Diseases.
- Ganley, K. J., Bosch, P. R., Blair, J. E., Rischard, F., & Galgiani, J. N. (2017). Oxygen Consumption Deficits in Patients With Residual Fatigue After Primary Coccidioidomycosis. Open Forum Infectious Diseases, 4(3). doi:https://doi.org/10.1093/ofid/ofx136
- Odio, C. D., Marciano, B. E., Galgiani, J. N., & Holland, S. M. (2017). Risk Factors for Disseminated Coccidioidomycosis, United States. Emerging infectious diseases, 23(2).More infoOf 150,000 new coccidioidomycosis infections that occur annually in the United States, ≈1% disseminate; one third of those cases are fatal. Immunocompromised hosts have higher rates of dissemination. We identified 8 patients with disseminated coccidioidomycosis who had defects in the interleukin-12/interferon-γ and STAT3 axes, indicating that these are critical host defense pathways.
- Galgiani, J. N., Ampel, N. M., Blair, J. E., Catanzaro, A., Geertsma, F., Hoover, S. E., Johnson, R. H., Kusne, S., Lisse, J., MacDonald, J. D., Meyerson, S. L., Raksin, P. B., Siever, J., Stevens, D. A., Sunenshine, R., & Theodore, N. (2016). 2016 Infectious Diseases Society of America (IDSA) Clinical Practice Guideline for the Treatment of Coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 63(6), e112-46.More infoIt is important to realize that guidelines cannot always account for individual variation among patients. They are not intended to supplant physician judgment with respect to particular patients or special clinical situations. Infectious Diseases Society of America considers adherence to these guidelines to be voluntary, with the ultimate determination regarding their application to be made by the physician in the light of each patient's individual circumstances.Coccidioidomycosis, also known as San Joaquin Valley fever, is a systemic infection endemic to parts of the southwestern United States and elsewhere in the Western Hemisphere. Residence in and recent travel to these areas are critical elements for the accurate recognition of patients who develop this infection. In this practice guideline, we have organized our recommendations to address actionable questions concerning the entire spectrum of clinical syndromes. These can range from initial pulmonary infection, which eventually resolves whether or not antifungal therapy is administered, to a variety of pulmonary and extrapulmonary complications. Additional recommendations address management of coccidioidomycosis occurring for special at-risk populations. Finally, preemptive management strategies are outlined in certain at-risk populations and after unintentional laboratory exposure.
- Galgiani, J. N., Ampel, N. M., Blair, J. E., Catanzaro, A., Geertsma, F., Hoover, S. E., Johnson, R. H., Kusne, S., Lisse, J., MacDonald, J. D., Meyerson, S. L., Raksin, P. B., Siever, J., Stevens, D. A., Sunenshine, R., & Theodore, N. (2016). Executive Summary: 2016 Infectious Diseases Society of America (IDSA) Clinical Practice Guideline for the Treatment of Coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 63(6), 717-22.More infoIt is important to realize that guidelines cannot always account for individual variation among patients. They are not intended to supplant physician judgment with respect to particular patients or special clinical situations. Infectious Diseases Society of America considers adherence to these guidelines to be voluntary, with the ultimate determination regarding their application to be made by the physician in the light of each patient's individual circumstances.Coccidioidomycosis, also known as San Joaquin Valley fever, is a systemic infection endemic to parts of the southwestern United States and elsewhere in the Western Hemisphere. Residence in and recent travel to these areas are critical elements for the accurate recognition of patients who develop this infection. In this practice guideline, we have organized our recommendations to address actionable questions concerning the entire spectrum of clinical syndromes. These can range from initial pulmonary infection, which eventually resolves whether or not antifungal therapy is administered, to a variety of pulmonary and extrapulmonary complications. Additional recommendations address management of coccidioidomycosis occurring for special at-risk populations. Finally, preemptive management strategies are outlined in certain at-risk populations and after unintentional laboratory exposure.
- Narra, H. P., Shubitz, L. F., Mandel, M. A., Trinh, H. T., Griffin, K., Buntzman, A. S., Frelinger, J. A., Galgiani, J. N., & Orbach, M. J. (2016). A Coccidioides posadasii CPS1 Deletion Mutant Is Avirulent and Protects Mice from Lethal Infection. Infection and immunity, 84(10), 3007-16.More infoThe CPS1 gene was identified as a virulence factor in the maize pathogen Cochliobolus heterostrophus Hypothesizing that the homologous gene in Coccidioides posadasii could be important for virulence, we created a Δcps1 deletion mutant which was unable to cause disease in three strains of mice (C57BL/6, BALB/c, or the severely immunodeficient NOD-scid,γc(null) [NSG]). Only a single colony was recovered from 1 of 60 C57BL/6 mice following intranasal infections of up to 4,400 spores. Following administration of very high doses (10,000 to 2.5 × 10(7) spores) to NSG and BALB/c mice, spherules were observed in lung sections at time points from day 3 to day 10 postinfection, but nearly all appeared degraded with infrequent endosporulation. Although the role of CPS1 in virulence is not understood, phenotypic alterations and transcription differences of at least 33 genes in the Δcps1 strain versus C. posadasii is consistent with both metabolic and regulatory functions for the gene. The in vitro phenotype of the Δcps1 strain showed slower growth of mycelia with delayed and lower spore production than C. posadasii, and in vitro spherules were smaller. Vaccination of C57BL/6 or BALB/c mice with live Δcps1 spores either intranasally, intraperitoneally, or subcutaneously resulted in over 95% survival with mean residual lung fungal burdens of
- Noble, J. A., Nelson, R. G., Fufaa, G. D., Kang, P., Shafir, S. C., & Galgiani, J. N. (2016). Effect of Geography on the Analysis of Coccidioidomycosis-Associated Deaths, United States. Emerging infectious diseases, 22(10), 1821-3.More infoBecause coccidioidomycosis death rates vary by region, we reanalyzed coccidioidomycosis-associated mortality in the United States by race/ethnicity, then limited analysis to Arizona and California. Coccidioidomycosis-associated deaths were shown to increase among African-Americans but decrease among Native Americans and Hispanics. Separately, in a Native American cohort, diabetes co-varied with coccidioidomycosis-associated death.
- Martirosyan, N. L., Skoch, J. M., Zaninovich, O., Zoccali, C., Galgiani, J. N., & Baaj, A. A. (2015). A paradigm for the evaluation and management of spinal coccidioidomycosis. Surgical neurology international, 6, 107.More infoCoccidioidomycosis is a fungal infection that is endemic to parts of the Southwestern United States. When infection involves the spine, the treatment strategies can be challenging. We have devised a management protocol for spinal coccidioidomycosis based on a review of the literature and our experience.
- Shubitz, L. F., Trinh, H. T., Galgiani, J. N., Lewis, M. L., Fothergill, A. W., Wiederhold, N. P., Barker, B. M., Lewis, E. R., Doyle, A. L., Hoekstra, W. J., Schotzinger, R. J., & Garvey, E. P. (2015). Evaluation of VT-1161 for Treatment of Coccidioidomycosis in Murine Infection Models. Antimicrobial agents and chemotherapy, 59(12), 7249-54.More infoCoccidioidomycosis, or valley fever, is a growing health concern endemic to the southwestern United States. Safer, more effective, and more easily administered drugs are needed especially for severe, chronic, or unresponsive infections. The novel fungal CYP51 inhibitor VT-1161 demonstrated in vitro antifungal activity, with MIC50 and MIC90 values of 1 and 2 μg/ml, respectively, against 52 Coccidioides clinical isolates. In the initial animal study, oral doses of 10 and 50 mg/kg VT-1161 significantly reduced fungal burdens and increased survival time in a lethal respiratory model in comparison with treatment with a placebo (P < 0.001). Oral doses of 25 and 50 mg/kg VT-1161 were similarly efficacious in the murine central nervous system (CNS) model compared to placebo treatment (P < 0.001). All comparisons with the positive-control drug, fluconazole at 50 mg/kg per day, demonstrated either statistical equivalence or superiority of VT-1161. VT-1161 treatment also prevented dissemination of infection from the original inoculation site to a greater extent than fluconazole. Many of these in vivo results can be explained by the long half-life of VT-1161 leading to sustained high plasma levels. Thus, the efficacy and pharmacokinetics of VT-1161 are attractive characteristics for long-term treatment of this serious fungal infection.
- Shubitz, L. F., Trinh, H. T., Powell, D. A., Lewis, M. L., Orbach, M. J., Frelinger, J. A., & Galgiani, J. N. (2017). Viable spores of Coccidioides posadasii delta-cps1 are required for vaccination and provide long lasting immunity. Vaccine.
- Wack, E. E., Ampel, N. M., Sunenshine, R. H., & Galgiani, J. N. (2015). The Return of Delayed-Type Hypersensitivity Skin Testing for Coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 61(5), 787-91.More infoA skin test that detects dermal hypersensitivity in persons with past infection with Coccidioides species is again available for clinical use. Nearly all of the clinical studies with similar materials were published prior to the 1990s, and as a result, many practicing physicians will be unfamiliar with how skin testing for coccidioidomycosis might be useful in patient management or as a research tool. We review clinical and epidemiological studies with past skin test antigens, the composition of past and current skin test preparations with particular attention to differences in the preservatives, and how the current preparation could be used today.
- Galgiani, J. N. (2014). 267 – Coccidioidomycosis (Coccidioides Species). Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases, 2974-2984. doi:10.1016/b978-1-4557-4801-3.00267-8
- Navalkar, K. A., Johnston, S. A., Woodbury, N., Galgiani, J. N., Magee, D. M., Chicacz, Z., & Stafford, P. (2014). Application of immunosignatures for diagnosis of valley fever. Clinical and vaccine immunology : CVI, 21(8), 1169-77.More infoValley fever (VF) is difficult to diagnose, partly because the symptoms of VF are confounded with those of other community-acquired pneumonias. Confirmatory diagnostics detect IgM and IgG antibodies against coccidioidal antigens via immunodiffusion (ID). The false-negative rate can be as high as 50% to 70%, with 5% of symptomatic patients never showing detectable antibody levels. In this study, we tested whether the immunosignature diagnostic can resolve VF false negatives. An immunosignature is the pattern of antibody binding to random-sequence peptides on a peptide microarray. A 10,000-peptide microarray was first used to determine whether valley fever patients can be distinguished from 3 other cohorts with similar infections. After determining the VF-specific peptides, a small 96-peptide diagnostic array was created and tested. The performances of the 10,000-peptide array and the 96-peptide diagnostic array were compared to that of the ID diagnostic standard. The 10,000-peptide microarray classified the VF samples from the other 3 infections with 98% accuracy. It also classified VF false-negative patients with 100% sensitivity in a blinded test set versus 28% sensitivity for ID. The immunosignature microarray has potential for simultaneously distinguishing valley fever patients from those with other fungal or bacterial infections. The same 10,000-peptide array can diagnose VF false-negative patients with 100% sensitivity. The smaller 96-peptide diagnostic array was less specific for diagnosing false negatives. We conclude that the performance of the immunosignature diagnostic exceeds that of the existing standard, and the immunosignature can distinguish related infections and might be used in lieu of existing diagnostics.
- Shubitz, L. F., Trinh, H. T., Perrill, R. H., Thompson, C. M., Hanan, N. J., Galgiani, J. N., & Nix, D. E. (2014). Modeling nikkomycin Z dosing and pharmacology in murine pulmonary coccidioidomycosis preparatory to phase 2 clinical trials. The Journal of infectious diseases, 209(12), 1949-54.More infoNikkomycin Z (NikZ) is a chitin synthase inhibitor with activity against Coccidioides species that is being developed as a first-in-class orphan product for treatment of coccidioidomycosis. It has previously been shown to reduce lethal respiratory infections in mice to undetectable levels when treatment is begun 48 hours after infection. The studies described here focus on bracketing NikZ doses for phase 2 and 3 clinical trials, using an established mouse respiratory infection as a model and starting treatment 120 hours after infection. A dose of 80 mg/kg/day, divided into 2 doses, nearly eradicated infection, and larger doses did not improve fungal clearance. Increasing the duration of treatment from 1 week to 3 weeks resulted in a greater percentage of culture-negative mice. Comparative data show that plasma levels of NikZ that nearly eradicate Coccidioides in mice are achievable in patients and provide a plausibly effective dose range for initial phase 2 clinical studies.
- Galgiani, J. N. (2013). Elements of style in managing coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 56(11), 1586-8.
- Nguyen, C., Barker, B. M., Hoover, S., Nix, D. E., Ampel, N. M., Frelinger, J. A., Orbach, M. J., & Galgiani, J. N. (2013). Recent advances in our understanding of the environmental, epidemiological, immunological, and clinical dimensions of coccidioidomycosis. Clinical microbiology reviews, 26(3), 505-25.More infoCoccidioidomycosis is the endemic mycosis caused by the fungal pathogens Coccidioides immitis and C. posadasii. This review is a summary of the recent advances that have been made in the understanding of this pathogen, including its mycology, genetics, and niche in the environment. Updates on the epidemiology of the organism emphasize that it is a continuing, significant problem in areas of endemicity. For a variety of reasons, the number of reported coccidioidal infections has increased dramatically over the past decade. While continual improvements in the fields of organ transplantation and management of autoimmune disorders and patients with HIV have led to dilemmas with concurrent infection with coccidioidomycosis, they have also led to advances in the understanding of the human immune response to infection. There have been some advances in therapeutics with the increased use of newer azoles. Lastly, there is an overview of the ongoing search for a preventative vaccine.
- Sampaio, E. P., Hsu, A. P., Pechacek, J., Bax, H. I., Dias, D. L., Paulson, M. L., Chandrasekaran, P., Rosen, L. B., Carvalho, D. S., Ding, L., Vinh, D. C., Browne, S. K., Datta, S., Milner, J. D., Kuhns, D. B., Long Priel, D. A., Sadat, M. A., Shiloh, M., De Marco, B., , Alvares, M., et al. (2013). Signal transducer and activator of transcription 1 (STAT1) gain-of-function mutations and disseminated coccidioidomycosis and histoplasmosis. The Journal of allergy and clinical immunology, 131(6), 1624-34.More infoImpaired signaling in the IFN-γ/IL-12 pathway causes susceptibility to severe disseminated infections with mycobacteria and dimorphic yeasts. Dominant gain-of-function mutations in signal transducer and activator of transcription 1 (STAT1) have been associated with chronic mucocutaneous candidiasis.
- Shubitz, L. F., Roy, M. E., Nix, D. E., & Galgiani, J. N. (2013). Efficacy of Nikkomycin Z for respiratory coccidioidomycosis in naturally infected dogs. Medical mycology, 51(7), 747-54.More infoNikkomycin Z (NikZ) is a chitin synthase inhibitor with antifungal efficacy against Coccidioides spp. and other endemic fungi. Dogs suffer a rate and range of natural coccidioidomycosis similar to humans and were considered an excellent model for initially testing NikZ against naturally acquired disease. Twelve dogs with coccidioidal pneumonia that had been present for an average of three months were treated with 250 mg (5-15 kg) or 500 mg (> 15-30 kg) twice daily for 60 days. Nine dogs completed the course of treatment and seven dogs had improvement in disease based on radiographs, clinicopathological parameters, physical examination findings, and subjective assessment by owners; three dogs had resolution or near resolution of disease. Based on this small study, NikZ shows efficacy to treat naturally acquired coccidioidomycosis and merits further development for trials in humans.
- Muhammed, M., Feldmesser, M., Shubitz, L. F., Lionakis, M. S., Sil, A., Wang, Y., Glavis-Bloom, J., Lewis, R. E., Galgiani, J. N., Casadevall, A., Kontoyiannis, D. P., & Mylonakis, E. (2012). Mouse models for the study of fungal pneumonia: a collection of detailed experimental protocols for the study of Coccidioides, Cryptococcus, Fusarium, Histoplasma and combined infection due to Aspergillus-Rhizopus. Virulence, 3(3), 329-38.More infoMouse models have facilitated the study of fungal pneumonia. In this report, we present the working protocols of groups that are working on the following pathogens: Aspergillus, Coccidioides, Cryptococcus, Fusarium, Histoplasma and Rhizopus. We describe the experimental procedures and the detailed methods that have been followed in the experienced laboratories to study pulmonary fungal infection; we also discuss the anticipated results and technical notes, and provide the practical advices that will help the users of these models.
- Stafford, P., Halperin, R., Legutki, J. B., Magee, D. M., Galgiani, J., & Johnston, S. A. (2012). Physical characterization of the "immunosignaturing effect". Molecular & cellular proteomics : MCP, 11(4), M111.011593.More infoIdentifying new, effective biomarkers for diseases is proving to be a challenging problem. We have proposed that antibodies may offer a solution to this problem. The physical features and abundance of antibodies make them ideal biomarkers. Additionally, antibodies are often elicited early in the ontogeny of different chronic and infectious diseases. We previously reported that antibodies from patients with infectious disease and separately those with Alzheimer's disease display a characteristic and reproducible "immunosignature" on a microarray of 10,000 random sequence peptides. Here we investigate the physical and chemical parameters underlying how immunosignaturing works. We first show that a variety of monoclonal and polyclonal antibodies raised against different classes of antigens produce distinct profiles on this microarray and the relative affinities are determined. A proposal for how antibodies bind the random sequences is tested. Sera from vaccinated mice and people suffering from a fugal infection are individually assayed to determine the complexity of signals that can be distinguished. Based on these results, we propose that this simple, general and inexpensive system could be optimized to generate a new class of antibody biomarkers for a wide variety of diseases.
- Taroumian, S., Knowles, S. L., Lisse, J. R., Yanes, J., Ampel, N. M., Vaz, A., Galgiani, J. N., & Hoover, S. E. (2012). Management of coccidioidomycosis in patients receiving biologic response modifiers or disease-modifying antirheumatic drugs. Arthritis care & research, 64(12), 1903-9.More infoCoccidioidomycosis (valley fever) is an endemic fungal infection of the American Southwest, an area with a large population of patients with rheumatic diseases. There are currently no guidelines for management of patients who develop coccidioidomycosis while under treatment with biologic response modifiers (BRMs) or disease-modifying antirheumatic drugs (DMARDs). We conducted a retrospective study of how both concurrent diseases were managed and the patient outcomes at 2 centers in Tucson, Arizona.
- Hector, R. F., Rutherford, G. W., Tsang, C. A., Erhart, L. M., McCotter, O., Anderson, S. M., Komatsu, K., Tabnak, F., Vugia, D. J., Yang, Y., & Galgiani, J. N. (2011). The public health impact of coccidioidomycosis in Arizona and California. International journal of environmental research and public health, 8(4), 1150-73.More infoThe numbers of reported cases of coccidioidomycosis in Arizona and California have risen dramatically over the past decade, with a 97.8% and 91.1% increase in incidence rates from 2001 to 2006 in the two states, respectively. Of those cases with reported race/ethnicity information, Black/African Americans in Arizona and Hispanics and African/Americans in California experienced a disproportionately higher frequency of disease compared to other racial/ethnic groups. Lack of early diagnosis continues to be a problem, particularly in suspect community-acquired pneumonia, underscoring the need for more rapid and sensitive tests. Similarly, the inability of currently available therapeutics to reduce the duration and morbidity of this disease underscores the need for improved therapeutics and a preventive vaccine.
- Shubitz, L. F., Dial, S. M., & Galgiani, J. N. (2011). T-lymphocyte predominance in lesions of canine coccidioidomycosis. Veterinary pathology, 48(5), 1008-11.More infoCoccidioidomycosis is a systemic fungal infection endemic to the southwestern United States. Although cell-mediated immunity is considered critical in control of the infection, little is known of the cellular population in naturally occurring lesions. To characterize the lymphocytic infiltration, archived formalin-fixed, paraffin-embedded tissues (subcutis, pericardium/heart, lung, bone, and synovium) from 18 dogs with coccidioidomycosis were studied with immunohistochemistry for CD3 and CD79a. In nearly all lesions, T lymphocytes were more numerous than B lymphocytes and were distributed throughout the lesion with concentration in the periphery of granulomas, whereas B lymphocytes were mostly confined to the periphery of granulomas. The predominance of T lymphocytes in lesions of canine coccidioidomycosis was independent of the tissue evaluated, the number of intralesional organisms, and the nature or severity of the inflammatory response.
- Shubitz, L., Dial, S. M., & Galgiani, J. N. (2011). T-lymphocyte predominance in lesions of canine coccidioidomycosis. Vet Pathol, 48(5), 1008-11. doi:10.1177/0300985810384410
- Vinh, D. C., Schwartz, B., Hsu, A. P., Miranda, D. J., Valdez, P. A., Fink, D., Lau, K. P., Long-Priel, D., Kuhns, D. B., Uzel, G., Pittaluga, S., Hoover, S., Galgiani, J. N., & Holland, S. M. (2011). Interleukin-12 receptor β1 deficiency predisposing to disseminated Coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 52(4), e99-e102.
- Galgiani, J. N., Shubitz, L. F., & Dial, S. M. (2010). T-Lymphocyte Predominance in Lesions of Canine Coccidioidomycosis. Veterinary Pathology, 48(5), 1008-1011. doi:10.1177/0300985810384410
- Neafsey, D. E., Barker, B. M., Sharpton, T. J., Stajich, J. E., Park, D. J., Whiston, E., Hung, C., McMahan, C., White, J., Sykes, S., Heiman, D., Young, S., Zeng, Q., Abouelleil, A., Aftuck, L., Bessette, D., Brown, A., FitzGerald, M., Lui, A., , Macdonald, J. P., et al. (2010). Population genomic sequencing of Coccidioides fungi reveals recent hybridization and transposon control. GENOME RESEARCH, 20(7), 938-946.
- Neafsey, D. E., Barker, B. M., Sharpton, T. J., Stajich, J. E., Park, D. J., Whiston, E., Hung, C., McMahan, C., White, J., Sykes, S., Heiman, D., Young, S., Zeng, Q., Abouelleil, A., Aftuck, L., Bessette, D., Brown, A., FitzGerald, M., Lui, A., , Macdonald, J. P., et al. (2010). Population genomic sequencing of Coccidioides fungi reveals recent hybridization and transposon control. Genome research, 20(7), 938-46.More infoWe have sequenced the genomes of 18 isolates of the closely related human pathogenic fungi Coccidioides immitis and Coccidioides posadasii to more clearly elucidate population genomic structure, bringing the total number of sequenced genomes for each species to 10. Our data confirm earlier microsatellite-based findings that these species are genetically differentiated, but our population genomics approach reveals that hybridization and genetic introgression have recently occurred between the two species. The directionality of introgression is primarily from C. posadasii to C. immitis, and we find more than 800 genes exhibiting strong evidence of introgression in one or more sequenced isolates. We performed PCR-based sequencing of one region exhibiting introgression in 40 C. immitis isolates to confirm and better define the extent of gene flow between the species. We find more coding sequence than expected by chance in the introgressed regions, suggesting that natural selection may play a role in the observed genetic exchange. We find notable heterogeneity in repetitive sequence composition among the sequenced genomes and present the first detailed genome-wide profile of a repeat-induced point mutation (RIP) process distinctly different from what has been observed in Neurospora. We identify promiscuous HLA-I and HLA-II epitopes in both proteomes and discuss the possible implications of introgression and population genomic data for public health and vaccine candidate prioritization. This study highlights the importance of population genomic data for detecting subtle but potentially important phenomena such as introgression.
- Ostrosky-Zeichner, L., Casadevall, A., Galgiani, J. N., Odds, F. C., & Rex, J. H. (2010). An insight into the antifungal pipeline: selected new molecules and beyond. NATURE REVIEWS DRUG DISCOVERY, 9(9), 719-727.
- Ostrosky-Zeichner, L., Casadevall, A., Galgiani, J. N., Odds, F. C., & Rex, J. H. (2010). An insight into the antifungal pipeline: selected new molecules and beyond. Nature reviews. Drug discovery, 9(9), 719-27.More infoInvasive fungal infections are increasing in incidence and are associated with substantial mortality. Improved diagnostics and the availability of new antifungals have revolutionized the field of medical mycology in the past decades. This Review focuses on recent developments in the antifungal pipeline, concentrating on promising candidates such as new azoles, polyenes and echinocandins, as well as agents such as nikkomycin Z and the sordarins. Developments in vaccines and antibody-based immunotherapy are also discussed. Few therapeutic products are currently in active development, and progression of therapeutic agents with fungus-specific mechanisms of action is of key importance.
- Stern, N. G., & Galgiani, J. N. (2010). Coccidioidomycosis among scholarship athletes and other college students, Arizona, USA. Emerging infectious diseases, 16(2), 321-3.More infoTo compare coccidioidomycosis case rates among groups of young adults in a disease-endemic region, we reviewed medical charts for serologic testing and coding. Case rates were higher for scholarship athletes than for other students and paralleled 5x more serologic testing. Our findings underscore the need to routinely test patients for coccidioidomycosis.
- Ampel, N. M., Dionne, S. O., Giblin, A., Podany, A. B., & Galgiani, J. (2009). Mannose-binding lectin serum levels are low in persons with clinically active coccidioidomycosis. Mycopathologia, 167(4), 173-80.More infoMannose-binding lectin (MBL) is a circulating collectin that is part of the innate immune response. We explored the serum levels of MBL in persons with different forms of coccidioidomycosis.
- Ampel, N. M., Giblin, A., Mourani, J. P., & Galgiani, J. N. (2009). Factors and outcomes associated with the decision to treat primary pulmonary coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 48(2), 172-8.More infoStudies that assess the value of initiating oral antifungal therapy to treat primary pulmonary coccidioidomycosis have not been published previously.
- Nix, D. E., Swezey, R. R., Hector, R., & Galgiani, J. N. (2009). Pharmacokinetics of nikkomycin Z after single rising oral doses. Antimicrobial agents and chemotherapy, 53(6), 2517-21.More infoNikkomycin Z is an antifungal drug that inhibits chitin synthase. This agent is under development as an orphan product for treatment of coccidioidomycosis. Safety and pharmacokinetics of nikkomycin Z were evaluated in healthy male subjects following single, rising oral doses ranging from 250 mg to 2,000 mg. A total of 12 subjects were recruited and divided into two groups. Group 1 (n = 6) received two out of three doses of 250 mg, 1,000 mg, or 1,750 mg and a placebo randomly in place of one of the doses. Group 2 (n = 6) received two out of three doses of 500 mg, 1,500 mg, or 2,000 mg and a placebo in place of one of the doses. Subjects were confined to the study unit overnight prior to dosing, and 12 blood samples were collected over 24 h postdosing while subjects were confined. Subjects returned for additional blood samples and safety evaluations at 48 h and 72 h after each dose. There was a 2-week washout period between doses. Plasma drug concentrations were determined using a validated high-performance liquid chromatography method. Nikkomycin Z was absorbed after oral administration, reaching a maximum concentration in serum of 2.21 microg/ml at 2 h postdose and an area under the concentration-time curve from 0 h to infinity of 11.3 microg x h/ml for the 250-mg dose. Pharmacokinetics appeared linear over the range of 250 to 500 mg; however, relative bioavailability was about 62 to 70% for the 1,000-mg dose and 42 to 47% for doses between 1,500 and 2,000 mg. The mean terminal half-life ranged from 2.1 to 2.5 h and was independent of dose. No serious or dose-related adverse events were observed. This study provides a basis for pharmacokinetic simulations and continued studies of nikkomycin Z administered in multiple doses.
- Sharpton, T. J., Stajich, J. E., Rounsley, S. D., Gardner, M. J., Wortman, J. R., Jordar, V. S., Maiti, R., Kodira, C. D., Neafsey, D. E., Zeng, Q., Hung, C., McMahan, C., Muszewska, A., Grynberg, M., Mandel, M. A., Kellner, E. M., Barker, B. M., Galgiani, J. N., Orbach, M. J., , Kirkland, T. N., et al. (2009). Comparative genomic analyses of the human fungal pathogens Coccidioides and their relatives. GENOME RESEARCH, 19(10), 1722-1731.
- Sharpton, T. J., Stajich, J. E., Rounsley, S. D., Gardner, M. J., Wortman, J. R., Jordar, V. S., Maiti, R., Kodira, C. D., Neafsey, D. E., Zeng, Q., Hung, C., McMahan, C., Muszewska, A., Grynberg, M., Mandel, M. A., Kellner, E. M., Barker, B. M., Galgiani, J. N., Orbach, M. J., , Kirkland, T. N., et al. (2009). Comparative genomic analyses of the human fungal pathogens Coccidioides and their relatives. Genome research, 19(10), 1722-31.More infoWhile most Ascomycetes tend to associate principally with plants, the dimorphic fungi Coccidioides immitis and Coccidioides posadasii are primary pathogens of immunocompetent mammals, including humans. Infection results from environmental exposure to Coccidiodies, which is believed to grow as a soil saprophyte in arid deserts. To investigate hypotheses about the life history and evolution of Coccidioides, the genomes of several Onygenales, including C. immitis and C. posadasii; a close, nonpathogenic relative, Uncinocarpus reesii; and a more diverged pathogenic fungus, Histoplasma capsulatum, were sequenced and compared with those of 13 more distantly related Ascomycetes. This analysis identified increases and decreases in gene family size associated with a host/substrate shift from plants to animals in the Onygenales. In addition, comparison among Onygenales genomes revealed evolutionary changes in Coccidioides that may underlie its infectious phenotype, the identification of which may facilitate improved treatment and prevention of coccidioidomycosis. Overall, the results suggest that Coccidioides species are not soil saprophytes, but that they have evolved to remain associated with their dead animal hosts in soil, and that Coccidioides metabolism genes, membrane-related proteins, and putatively antigenic compounds have evolved in response to interaction with an animal host.
- Vinh, D. C., Masannat, F., Dzioba, R. B., Galgiani, J. N., & Holland, S. M. (2009). Refractory disseminated coccidioidomycosis and mycobacteriosis in interferon-gamma receptor 1 deficiency. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 49(6), e62-5.More infoSevere coccidioidomycosis is rare, and specific genetic susceptibility to the disease remains unidentified. We describe a patient with disseminated recalcitrant coccidioidomycosis with autosomal dominant interferon-gamma receptor 1 deficiency caused by a heterozygous IFNGR1 818del4 mutation. Therefore, the interleukin-12/interferon-gamma axis appears to be critical for control of coccidioidomycosis.
- Galgiani, J. N. (2008). Vaccines to prevent systemic mycoses: holy grails meet translational realities. The Journal of infectious diseases, 197(7), 938-40.
- Galgiani, J. N., Shubitz, L. F., Dial, S. M., Perrill, R., & Casement, R. (2008). Vaccine-Induced Cellular Immune Responses Differ from Innate Responses in Susceptible and Resistant Strains of Mice Infected with Coccidioides posadasii. Infection and Immunity, 76(12), 5553-5564. doi:10.1128/iai.00885-08
- Shubitz, L. F., Dial, S. M., Perrill, R., Casement, R., & Galgiani, J. N. (2008). Vaccine-induced cellular immune responses differ from innate responses in susceptible and resistant strains of mice infected with Coccidioides posadasii. Infection and immunity, 76(12), 5553-64.More infoSusceptibility to Coccidioides spp. varies widely in humans and other mammals and also among individuals within a species. Among strains of mice with various susceptibilities, immunohistopathology revealed that C57BL/6 mice were highly susceptible to the disease following intranasal infection, DBA/2n mice were intermediate, and Swiss-Webster mice were innately resistant. Resistant Swiss-Webster mice developed prominent perivascular/peribronchiolar lymphocytic cuffing and well-formed granulomas with few fungal elements and debris in the necrotic center, surrounded by a mantle of macrophages, lymphocytes, and fibrocytes. Susceptible C57BL/6 mice became moribund between 14 and 18 days postinfection, with overwhelming numbers of neutrophils and spherules and very few T cells, the drastic reduction of which was associated with failure and death, while intermediate DBA/2n mice controlled the fungal burden but demonstrated progressive lung inflammation with prominent suppuration, and they deteriorated clinically. Vaccinated C57BL/6 mice had an early and robust lymphocyte response, which included significantly higher Mac2(+), CD3(+), and CD4(+) cell scores on day 18 than those of innately resistant SW mice and DBA/2n mice; they also had prominent perivascular/peribronchiolar lymphocytic infiltrates not present in their unvaccinated counterparts, and they appeared to be resolving lesions by day 56 compared to the other two strains, based on significantly lower disease scores and observably smaller and fewer lesions with few spherules and neutrophils.
- Shubitz, L., Dial, S. M., Perrill, R., Casement, R., & Galgiani, J. N. (2008). Vaccine-induced cellular immune responses differ from innate responses in susceptible and resistant strains of mice infected with Coccidioides posadasii.. Infect Immun, 76(12), 5553-5564. doi:10.1128/IAI.00885-08
- Catanzaro, A., Cloud, G. A., Stevens, D. A., Levine, B. E., Williams, P. L., Johnson, R. H., Rendon, A., Mirels, L. F., Lutz, J. E., Holloway, M., & Galgiani, J. N. (2007). Safety, tolerance, and efficacy of posaconazole therapy in patients with nonmeningeal disseminated or chronic pulmonary coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 45(5), 562-8.More infoCoccidioidomycosis can be difficult to treat with available therapies, particularly in patients with progressive or disseminated disease. Posaconazole is a new azole antifungal with potent activity against Coccidioides species, the causative agent of coccidioidomycosis.
- Galgiani, J. N. (2007). Coccidioidomycosis: changing perceptions and creating opportunities for its control. Annals of the New York Academy of Sciences, 1111, 1-18.More infoThe perceptions of coccidioidomycosis as a medical problem has undergone sequential and dramatic metamorphoses since its first description more than a century ago. First thought to be rare and lethal, coccidioidomycosis was subsequently found to be common and often mild. During World War II, its overall impact upon large populations came sharply into focus and the consequences for public health became clearer. Early treatments had significant limitations and toxicities, and therefore treatment of coccidioidomycosis was reserved for only the sickest patients. Since then, safer oral therapies have become commonplace. Despite their availability, there has been no investigation of their use in the less severe and much more common early infections. Even newer drugs such as nikkomycin Z, which might actually cure infections, until very recently have had trouble finding a sponsor to move it through clinical trials. Perceptions once formed by the understanding of coccidioidomycosis as a medical problem now appear to hinder the future study of newer therapeutic opportunities. It is suggested in this review that it is time to revisit and possibly change these perceptions if we are to improve our care of patients.
- Johnson, S. M., Lerche, N. W., Pappagianis, D., Yee, J. L., Galgiani, J. N., & Hector, R. F. (2007). Safety, antigenicity, and efficacy of a recombinant coccidioidomycosis vaccine in cynomolgus macaques (Macaca fascicularis). Annals of the New York Academy of Sciences, 1111, 290-300.More infoThe safety, immunogenicity and efficacy of recombinant Ag2/PRA106 + CSA chimeric fusion protein (CFP) vaccine in ISS/Montanide adjuvant-administered intramuscular (IM) was assessed in adult female cynomolgus macaques challenged with Coccidioides posadasii. Animals received three immunizations with either 5 microg CFP, 50-microg CFP, or adjuvant alone and were challenged 4 weeks following the final immunization. Although significant antibody response was produced in response to vaccination, there were no discernable adverse effects, suggesting that the vaccine was well tolerated. Upon intratracheal challenge, all animals showed evidence of disease. Two animals that received 5-microg doses of CFP were euthanatized prior to the study's end because of severe symptoms. Animals vaccinated with 50-microg doses of CFP showed evidence of enhanced sensitization compared to adjuvant controls and animals vaccinated with 5-microg doses of CFP. This was based on higher serum anti-CFP titers, enhanced secretion of interferon-gamma (IFN-gamma) from stimulated bronchoalveolar lavage mononuclear cells (BALMC), reduced pulmonary radiologic findings following intratracheal challenge, reduced terminal complement fixation titers, and reduced necropsy findings. Overall the vaccine was well tolerated, induced sensitization, and resulted in a protective response when given at the higher 50-microg dose. Additional experiments may be needed to optimize the vaccination and to confer greater protection against lethal challenge.
- Li, L., Schmelz, M., Kellner, E. M., Galgiani, J. N., & Orbach, M. J. (2007). Nuclear labeling of Coccidioides posadasii with green fluorescent protein. Annals of the New York Academy of Sciences, 1111, 198-207.More infoCoccidioidomycosis is a mild to life-threatening disease in otherwise healthy humans and other mammals caused by the fungus Coccidioides spp. Understanding the development of the unique dimorphic life cycle of Coccidioides spp. and its role in pathogenesis has been an area of research focus. However, nuclear behavior during the saprobic and parasitic life cycle has not been studied intensively. In this study, green fluorescent protein (GFP) was fused to histone H1 and introduced into Coccidioides posadasii (C. posadasii) strain Silveira to monitor the nuclear behavior of the fungus during the saprobic and parasitic stages of the life cycle. We constructed an Agrobacterium tumefaciens-mediated transformation (ATMT) vector that had in its T-DNA region a hygromycin-resistance gene as well as the fused histone H1-GFP gene under the control of the histone H3 promoter of C. posadasii. More than 30 hygromycin-resistant transformants were obtained and 23 were purified to homozygosity through multiple passages of the original transformants on hygromycin-containing media. One strain (VFC1420) transformed with a single copy of the fusion histone H1-GFP gene was selected for cytological studies. Strong nuclear-localized GFP signals were observed in arthroconidia, hyphae, as well as in spherules and endospores developed in vitro. Thus GFP can be used to study the expression pattern of potential virulence genes identified in serial analysis of gene expression (SAGE) or expressed sequence tags (EST) libraries, and could be a useful tool to monitor disease development in the murine model.
- Rohrbough, J. G., Galgiani, J. N., & Wysocki, V. H. (2007). The application of proteomic techniques to fungal protein identification and quantification. Annals of the New York Academy of Sciences, 1111, 133-46.More infoThe number of sequenced genomes has increased rapidly in the last few years, supporting a revolution in bioinformatics that has been leveraged by scientists seeking to analyze the proteomes of numerous biological systems. The primary technique employed for the identification of peptides and proteins from biological sources is mass spectrometry (MS). This analytical process is usually in the form of whole-protein analysis (termed "top-down" proteomics) or analysis of enzymatically produced peptides (known as the "bottom-up" approach). This article will focus primarily on the more common bottom-up proteomics to include topics such as sample preparation, separation strategies, MS instrumentation, data analysis, and techniques for protein quantification. Strategies for preparation of samples for proteomic analysis, as well as tools for protein and peptide separation will be discussed. A general description of common MS instruments along with tandem mass spectrometry (MS/MS) will be given. Different methodologies of sample ionization including matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) will be discussed. Data analysis methods including database search algorithms and tools for protein sequence analysis will be introduced. We will also discuss experimental strategies for MS protein quantification using stable isotope labeling techniques and fluorescent labeling. We will introduce several fungal proteomic studies to illustrate the use of these methods. This article will allow investigators to gain a working knowledge of proteomics along with some strengths and weaknesses associated with the techniques presented.
- Dionne, S. O., Podany, A. B., Ruiz, Y. W., Ampel, N. M., Galgiani, J. N., & Lake, D. F. (2006). Spherules derived from Coccidioides posadasii promote human dendritic cell maturation and activation. Infection and immunity, 74(4), 2415-22.More infoPrevious studies have shown that dendritic cells (DC) pulsed with T27K, an antigenic preparation derived from spherules (of Coccidioides posadasii), activate peripheral blood mononuclear cells (PBMC) from nonimmune subjects as well as from patients with disseminated coccidioidomycosis. In this study, we have assessed the interaction between human DC and C. posadasii spherules in order to better understand the initial response between Coccidioides and the human host. Whole autoclaved spherules induced lymphocyte transformation in PBMC obtained from immune but not from nonimmune donors. Immature DC (iDC) bound fluorescein isothiocyanate-labeled spherules in a time- and temperature-dependent manner. This binding was blocked by the addition of mannan, suggesting mannose receptor involvement in the DC-Coccidioides interaction. Binding was subsequently associated with ingestion and intracellular processing of spherules. Coculturing of spherules with iDC was associated with the development of mature DC that were morphologically, phenotypically, and functionally similar to those induced by tumor necrosis factor alpha and prostaglandin E2. Finally, spherules incubated with iDC induced activation of PBMC from nonimmune donors. These data indicate that human DC are capable of binding, internalizing, and presenting antigens from Coccidioides spherules and suggest that DC may play a critical early role in the formation of a cellular immune response in human coccidioidomycosis.
- Mandel, M. A., Galgiani, J. N., Kroken, S., & Orbach, M. J. (2006). Coccidioides posadasii contains single chitin synthase genes corresponding to classes I to VII. Fungal genetics and biology : FG & B, 43(11), 775-88.More infoCoccidioides posadasii is a dimorphic fungal pathogen of humans and other mammals. The switch between saprobic and parasitic growth involves synthesis of new cell walls of which chitin is a significant component. To determine whether particular subsets of chitin synthases (CHSes) are responsible for production of chitin at different stages of differentiation, we have isolated six CHS genes from this fungus. They correspond, together with another reported CHS gene, to single members of the seven defined classes of chitin synthases (classes I-VII). Using Real-Time RT-PCR we show their pattern of expression during morphogenesis. CpCHS2, CpCHS3, and CpCHS6 are preferentially expressed during the saprobic phase, while CpCHS1 and CpCHS4 are more highly expressed during the parasitic phase. CpCHS5 and CpCHS7 expression is similar in both saprobic and parasitic phases. Because C. posadasii contains single members of the seven classes of CHSes found in fungi, it is a good model to investigate the putatively different roles of these genes in fungal growth and differentiation.
- Orsborn, K. I., Shubitz, L. F., Peng, T., Kellner, E. M., Orbach, M. J., Haynes, P. A., & Galgiani, J. N. (2006). Protein expression profiling of Coccidioides posadasii by two-dimensional differential in-gel electrophoresis and evaluation of a newly recognized peroxisomal matrix protein as a recombinant vaccine candidate. Infection and immunity, 74(3), 1865-72.More infoCoccidioides posadasii and Coccidioides immitis are dimorphic, soil-dwelling pathogenic ascomycetes endemic to the southwestern United States. Infection can result from inhalation of a very few arthroconidia, but following natural infection, long-lived immunity is the norm. Previous work in the field has shown that spherule-derived vaccines afford more protection than those from mycelia. We have used two-dimensional differential in-gel electrophoresis coupled with nano-high-performance liquid chromatography-tandem mass spectrometry to directly assess both absolute abundance and differential expression of proteins in the spherule and the mycelial phases of C. posadasii with the intent to identify potential vaccine candidates. Peptides derived from 40 protein spots were analyzed and a probable identity was assigned to each. One spherule-abundant protein, identified as Pmp1, showed homology to allergens from Aspergillus fumigatus and other fungi, all of which exhibit similarity to yeast thiol peroxidases. Recombinant Pmp1 was reactive with serum from individuals with both acute and protracted disease, and evoked protection in two murine models of infection with C. posadasii. These results demonstrate the utility of proteomic analysis as a point of discovery for protective antigens for possible inclusion in a vaccine candidate to prevent coccidioidomycosis.
- Shubitz, L. F., Galgiani, J. N., Tian, Z., Zhong, Z., Timmermans, P., & Katz, L. (2006). Efficacy of ambruticin analogs in a murine model of coccidioidomycosis. Antimicrobial agents and chemotherapy, 50(10), 3467-9.More infoAmbruticin S, an antifungal cyclopropyl-pyran acid, showed curative effects against murine coccidioidal infection. Two analogs of this compound with greater in vitro potency were tested against lethal murine Coccidioides infection. Both improved the survival of mice over that of controls; one resulted in near-sterilization of infection.
- Shubitz, L. F., Yu, J., Hung, C., Kirkland, T. N., Peng, T., Perrill, R., Simons, J., Xue, J., Herr, R. A., Cole, G. T., & Galgiani, J. N. (2006). Improved protection of mice against lethal respiratory infection with Coccidioides posadasii using two recombinant antigens expressed as a single protein. Vaccine, 24(31-32), 5904-11.More infoTwo recombinant antigens which individually protect mice from lethal intranasal infection were studied in combination, either as a mixture of two separately expressed proteins or as a single chimeric expression product. Mice vaccinated with either combination survived longer than mice given single antigens. Immunized mice also exhibited specific IgG immunoglobulins and yielded splenocytes which produced interferon-gamma in response to either antigen. The chimeric antigen has the practical advantage of offering enhanced protection from multiple components without increasing production costs.
- Valdivia, L., Nix, D., Wright, M., Lindberg, E., Fagan, T., Lieberman, D., Stoffer, T., Ampel, N. M., & Galgiani, J. N. (2006). Coccidioidomycosis as a common cause of community-acquired pneumonia. Emerging infectious diseases, 12(6), 958-62.More infoThe early manifestations of coccidioidomycosis (valley fever) are similar to those of other causes of community-acquired pneumonia (CAP). Without specific etiologic testing, the true frequency of valley fever may be underestimated by public health statistics. Therefore, we conducted a prospective observational study of adults with recent onset of a lower respiratory tract syndrome. Valley fever was serologically confirmed in 16 (29%) of 55 persons (95% confidence interval 16%-44%). Antimicrobial medications were used in 81% of persons with valley fever. Symptomatic differences at the time of enrollment had insufficient predictive value for valley fever to guide clinicians without specific laboratory tests. Thus, valley fever is a common cause of CAP after exposure in a disease-endemic region. If CAP develops in persons who travel or reside in Coccidioides-endemic regions, diagnostic evaluation should routinely include laboratory evaluation for this organism.
- Galgiani, J. N., Ampel, N. M., Blair, J. E., Catanzaro, A., Johnson, R. H., Stevens, D. A., & Williams, P. L. (2005). Coccidioidomycosis. CLINICAL INFECTIOUS DISEASES, 41(9), 1217-1223.
- Galgiani, J. N., Ampel, N. M., Blair, J. E., Catanzaro, A., Johnson, R. H., Stevens, D. A., Williams, P. L., & , I. D. (2005). Coccidioidomycosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 41(9), 1217-23.
- Kellner, E. M., Orsborn, K. I., Siegel, E. M., Mandel, M. A., Orbach, M. J., & Galgiani, J. N. (2005). Coccidioides posadasii contains a single 1,3-beta-glucan synthase gene that appears to be essential for growth. Eukaryotic cell, 4(1), 111-20.More info1,3-beta-Glucan synthase is responsible for the synthesis of beta-glucan, an essential cell wall structural component in most fungi. We sought to determine whether Coccidioides posadasii possesses genes homologous to known fungal FKS genes that encode the catalytic subunit of 1,3-beta-glucan synthase. A single gene, designated FKS1, was identified, and examination of its predicted protein product showed a high degree of conservation with Fks proteins from other filamentous fungi. FKS1 is expressed at similar levels in mycelia and early spherulating cultures, and expression decreases as the spherules mature. We used Agrobacterium-mediated transformation to create strains that harbor DeltaFKS1::hygB, a null allele of FKS1, and hypothesize that Fks1p function is essential, due to our inability to purify this allele away from a complementing wild-type FKS1 allele in a heterokaryotic strain. The heterokaryon appears normal with respect to growth rate and arthroconidium production; however, microscopic examination of strains with DeltaFKS1::hygB alleles revealed abnormal swelling of hyphal elements.
- Park, B. J., Sigel, K., Vaz, V., Komatsu, K., McRill, C., Phelan, M., Colman, T., Comrie, A. C., Warnock, D. W., Galgiani, J. N., & Hajjeh, R. A. (2005). An epidemic of coccidioidomycosis in Arizona associated with climatic changes, 1998-2001. The Journal of infectious diseases, 191(11), 1981-7.More infoReports of coccidioidomycosis cases in Arizona have increased substantially. We investigated factors associated with the increase.
- Bennett, J. E., Kauffman, C., Walsh, T., de Pauw, B., Dismukes, W., Galgiani, J., Glauser, M., Herbrecht, R., Lee, J., Pappas, P., Powers, J., Rex, J., Verweij, P., & Viscoli, C. (2003). Forum report: issues in the evaluation of diagnostic tests, use of historical controls, and merits of the current multicenter collaborative groups. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 36(Suppl 3), S123-7.More infoThis forum report contains conclusions about 3 different issues relevant to conducting clinical trials in deep mycoses. (1) Trials of diagnostic tests for deep mycoses must define the population appropriate for testing and the clinical question being asked. The unanswered question for the serum Aspergillus galactomannan assay is whether knowledge of results can change use of empirical therapy to treat febrile patients at high risk of invasive aspergillosis. (2) Use of historical controls is suboptimal but offers a pragmatic solution for studying rare mycoses; use of contemporaneous controls, matched for critical variables and evaluated by a blinded data review committee using detailed criteria, appears optimal. (3) Established groups of independent investigators, such as the European Organization for Research on Treatment of Cancer's Invasive Fungal Infections Group and National Institute of Allergy and Infectious Diseases's Bacteriology and Mycology Study Group, provide a pool of experienced investigators, defined operating rules, impartiality, and specialized expertise. Considering the enormous investment required for adequately powered efficacy trials of antifungal agents and the importance of these trials to guide clinical practice, use of collaborative groups outweighs the extra administrative time that is sometimes required.
- Bennett, J. E., Powers, J., Walsh, T., Viscoli, C., de Pauw, B., Dismukes, W., Galgiani, J., Glauser, M., Herbrecht, R., Kauffman, C., Lee, J., Pappas, P., Rex, J., & Verweij, P. (2003). Forum report: issues in clinical trials of empirical antifungal therapy in treating febrile neutropenic patients. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 36(Suppl 3), S117-22.More infoThere is inferential evidence that some patients with prolonged neutropenia and fever not responding to antibacterial agents are at sufficient risk of deep mycoses to warrant empirical therapy, although superiority of an antifungal agent over placebo has not been conclusively demonstrated. Amphotericin B deoxycholate, liposomal amphotericin B, and intravenous itraconazole followed by oral itraconazole solution are licensed in the United States for this indication. Fluconazole and voriconazole have given favorable results in clinical trials of patients with low and high risk of deep mold infections, respectively. Design features that can profoundly influence outcome of empirical trials are (1) inclusion of low-risk patients, (2) failure to blind the study, (3) obscuration of antifungal effects by changing antibacterial antibiotics, (4) failure to balance both arms of the study in terms of patients with prior antifungal prophylaxis or with severe comorbidities, (5) the merging of end points evaluating safety with those of efficacy, and (6) choice of different criteria for resolution of fever.
- Bennett, J. E., Powers, J., de Pauw, B., Dismukes, W., Galgiani, J., Glauser, M., Herbrecht, R., Kauffman, C., Lee, J., Pappas, P., Rex, J., Verweij, P., Viscoli, C., & Walsh, T. (2003). Forum report: issues in the design of trials of drugs for the treatment of invasive aspergillosis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 36(Suppl 3), S113-6.More infoA recent trial of drugs for invasive aspergillosis was used as a background for discussing critical features in the design of antifungal trials. The study under discussion allowed stopping either drug without classifying the patient as having treatment failure, so the trial should be understood as a comparison of 2 treatment strategies, not just 2 drugs. Although the study was a noninferiority trial, the outcome permitted a claim of superiority. Use of the category of "probable" in addition to "proven" aspergillosis permitted inclusion of patients for whom the diagnosis was less certain but who were still early enough in the disease progression to respond to therapy. Different opinions still exist about some of the criteria for the diagnosis of "probable" aspergillosis. A blinded data review committee was helpful in evaluating efficacy in this unblinded trial but had limited value in assessing toxicity. An understanding of these features of design of antifungal drug trials is important in applying the results to clinical practice.
- Chiller, T. M., Galgiani, J. N., & Stevens, D. A. (2003). Coccidioidomycosis. INFECTIOUS DISEASE CLINICS OF NORTH AMERICA, 17(1), 41-+.
- Chiller, T. M., Galgiani, J. N., & Stevens, D. A. (2003). Coccidioidomycosis. Infectious disease clinics of North America, 17(1), 41-57, viii.More infoCoccidioides, a fungus, is endemic to specific parts of the Western Hemisphere. This article examines the prevalence, pathogenesis and host defense, clinical manifestations, diagnosis, and treatment of coccidioidomycosis.
- Abuodeh, R. O., Galgiani, J. N., & Scalarone, G. M. (2002). Molecular approaches to the study of Coccidioides immitis. International journal of medical microbiology : IJMM, 292(5-6), 373-80.More infoThe study of the molecular biology of Coccidioides sp. is only just beginning. As the importance of coccidioidomycosis grows as a public health problem, our need for understanding of pathogenesis, immune responses, and improved antifungal therapy also increases in proportion. Tools have now become available to study gene manipulation in this pathogen and this will allow molecular approaches to be used. Genetic experiments will also be accelerated by the availability of the whole coccidioidal genome, expected to be made public in the spring of 2003 (see http://www.tigr.org/tdb/tgi/cigi/GenInfo.html). Thus, there seems to be several reasons to expect considerable progress in the coming years.
- Peng, T., Shubitz, L., Simons, J., Perrill, R., Orsborn, K. I., & Galgiani, J. N. (2002). Localization within a proline-rich antigen (Ag2/PRA) of protective antigenicity against infection with Coccidioides immitis in mice. Infection and immunity, 70(7), 3330-5.More infoSubunits of a proline-rich coccidioidal antigen (Ag2/PRA) of Coccidioides immitis were analyzed by comparison as vaccines in mice. The optimal dose of plasmid vaccine encoding full-length Ag2/PRA was determined to be between 10 and 100 microg. Mice vaccinated with plasmids encoding amino acids (aa) 1 to 106 were as protective as full-length Ag2/PRA (aa 1 to 194). The subunit from aa 27 to 106 was significantly but less protective. Plasmids encoding aa 90 to 151 or aa 90 to 194 were not protective. Analogous results were obtained with recombinant vaccines of the same amino acid sequences. In addition, mixtures of aa 90 to 194 with either aa 1 to 106 or aa 27 to 106 did not enhance protection compared to the active single-recombinant subunits alone. Humoral response of total immunoglobulin G (IgG) and subclasses IgG1 and IgG2a were detectable in subunit vaccinations but at significantly (100-fold) lower concentrations than after vaccination with plasmids encoding full-length Ag2/PRA. Since virtually all protection by vaccination with full-length Ag2/PRA can be accounted for in the first half of the protein (aa 1 to 106), this subunit could make a multicomponent vaccine more feasible by reducing the quantity of protein per dose and the possibility of an untoward reactions to a foreign protein.
- Shubitz, L., Peng, T., Perrill, R., Simons, J., Orsborn, K., & Galgiani, J. N. (2002). Protection of mice against Coccidioides immitis intranasal infection by vaccination with recombinant antigen 2/PRA. Infection and immunity, 70(6), 3287-9.More infoSubcutaneous vaccination with recombinant antigen 2/PRA (rAg2/PRA) protected BALB/c mice against intranasal infection with Coccidioides immitis. Subcutaneously vaccinated C57BL/6 mice and intranasally vaccinated BALB/c mice were protected against larger numbers of infecting spores. Weight loss correlated with lethality, but histologic appearance did not. These studies support rAg2/PRA vaccination to prevent coccidioidomycosis.
- Galgiani, J. N., Ampel, N. M., Catanzaro, A., Johnson, R. H., Stevens, D. A., & Williams, P. L. (2000). Practice guidelines for the treatment of coccidioidomycosis. CLINICAL INFECTIOUS DISEASES, 30(4), 658-661.
- Galgiani, J. N., Schafer, F., Orsborn, K. I., Orbach, M. J., Murphy, J. W., Mandel, M. A., Grace, G. G., & Galgiani, J. N. (2000). The Cryptococcus neoformans gene DHA1 encodes an antigen that elicits a delayed-type hypersensitivity reaction in immune mice.. Infection and immunity, 68(11), 6196-201. doi:10.1128/iai.68.11.6196-6201.2000More infoWhen mice are vaccinated with a culture filtrate from Cryptococcus neoformans (CneF), they mount a protective cell-mediated immune response as detected by dermal delayed-type hypersensitivity (DTH) to CneF. We have identified a gene (DHA1) whose product accounts at least in part for the DTH reactivity. Using an acapsular mutant (Cap-67) of C. neoformans strain B3501, we prepared a culture filtrate (CneF-Cap67) similar to that used for preparing the commonly used skin test antigen made with C. neoformans 184A (CneF-184A). CneF-Cap67 elicited DTH in mice immunized with CneF-184A. Deglycosylation of CneF-Cap67 did not diminish its DTH activity. Furthermore, size separation by either chromatography or differential centrifugation identified the major DTH activity of CneF-Cap67 to be present in fractions that contained proteins of approximately 19 to 20 kDa. Using N-terminal and internal amino acid sequences derived from the 20-kDa band, oligonucleotide primers were designed, two of which produced a 776-bp amplimer by reverse transcription-PCR (RT-PCR) using RNA from Cap-67 to prepare cDNA for the template. The amplimer was used as a probe to isolate clones containing the full-length DHA1 gene from a phage genomic library prepared from strain B3501. The full-length cDNA was obtained by 5' rapid amplification of cDNA ends and RT-PCR. Analysis of DHA1 revealed a similarity between the deduced open reading frame and that of a developmentally regulated gene from Lentinus edodes (shiitake mushroom) associated with fruiting-body formation. Also, the gene product contained several amino acid sequences identical to those determined biochemically from the purified 20-kDa peptide encoded by DHA1. Recombinant DHA1 protein expressed in Escherichia coli was shown to elicit DTH reactions similar to those elicited by CneF-Cap67 in mice immunized against C. neoformans. Thus, DHA1 is the first gene to be cloned from C. neoformans whose product has been shown to possess immunologic activity.
- Rex, J. H., Pfaller, M. A., Galgiani, J. N., Bartlett, M. S., EspinelIngroff, A., Ghannoum, M. A., Lancaster, M., Odds, F. C., Rinaldi, M. G., Walsh, T. J., & Barry, A. L. (1997). Development of interpretive breakpoints for antifungal susceptibility testing: Conceptual framework and analysis of in vitro in vivo correlation data for fluconazole, itraconazole, and Candida infections. CLINICAL INFECTIOUS DISEASES, 24(2), 235-247.
- Fridkin, S. K., Pear, S. M., Williamson, T. H., Galgiani, J. N., & Jarvis, W. R. (1996). The role of understaffing in central venous catheter-associated bloodstream infections. INFECTION CONTROL AND HOSPITAL EPIDEMIOLOGY, 17(3), 150-158.
- Ghannoum, M. A., Rex, J. H., & Galgiani, J. N. (1996). Susceptibility testing of fungi: Current status of correlation of in vitro data with clinical outcome. JOURNAL OF CLINICAL MICROBIOLOGY, 34(3), 489-495.
- REX, J. H., COOPER, C. R., MERZ, W. G., GALGIANI, J. N., & ANAISSIE, E. J. (1995). DETECTION OF AMPHOTERICIN B-RESISTANT CANDIDA ISOLATES IN A BROTH-BASED SYSTEM. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 39(4), 906-909.
- DENNING, D. W., LEE, J. Y., HOSTETLER, J. S., PAPPAS, P., KAUFFMAN, C. A., DEWSNUP, D. H., GALGIANI, J. N., GRAYBILL JR, ., SUGAR, A. M., CATANZARO, A., GALLIS, H., PERFECT JR, ., DOCKERY, B., DISMUKES, W. E., & STEVENS, D. A. (1994). NIAID MYCOSES STUDY-GROUP MULTICENTER TRIAL OF ORAL ITRACONAZOLE THERAPY FOR INVASIVE ASPERGILLOSIS. AMERICAN JOURNAL OF MEDICINE, 97(2), 135-144.
- PEAR, S. M., WILLIAMSON, T. H., BETTIN, K. M., GERDING, D. N., & GALGIANI, J. N. (1994). DECREASE IN NOSOCOMIAL CLOSTRIDIUM-DIFFICILE-ASSOCIATED DIARRHEA BY RESTRICTING CLINDAMYCIN USE. ANNALS OF INTERNAL MEDICINE, 120(4), 272-277.
- FROMTLING, R. A., GALGIANI, J. N., PFALLER, M. A., ESPINELINGROFF, A., BARTIZAL, K. F., BARTLETT, M. S., BODY, B. A., FREY, C., HALL, G., ROBERTS, G. D., NOLTE, F. B., ODDS, F. C., RINALDI, M. G., SUGAR, A. M., & VILLAREAL, K. (1993). MULTICENTER EVALUATION OF A BROTH MACRODILUTION ANTIFUNGAL SUSCEPTIBILITY TEST FOR YEASTS. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 37(1), 39-45.
- GALGIANI, J. N., CATANZARO, A., CLOUD, G. A., HIGGS, J., FRIEDMAN, B. A., LARSEN, R. A., & GRAYBILL JR, . (1993). FLUCONAZOLE THERAPY FOR COCCIDIOIDAL MENINGITIS. ANNALS OF INTERNAL MEDICINE, 119(1), 28-35.
- REX, J. H., PFALLER, M. A., RINALDI, M. G., POLAK, A., & GALGIANI, J. N. (1993). ANTIFUNGAL SUSCEPTIBILITY TESTING. CLINICAL MICROBIOLOGY REVIEWS, 6(4), 367-381.
- Vincent, T., Galgiani, J. N., Huppert, M., & Salkin, D. (1993). The natural history of coccidioidal meningitis: VA-Armed Forces cooperative studies, 1955-1958.. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 16(2), 247-54. doi:10.1093/clind/16.2.247More infoFrom records on the clinical course of 699 military and veteran patients who had coccidioidomycosis before the advent of effective antifungal therapy, we identified 25 cases in which the manifestations of meningeal dissemination were described. Of 21 patients who had an identifiable initial infection, 16 developed meningeal symptoms (most frequently headache, vomiting, and nuchal rigidity) within the next 6 months. Associated pulmonary lesions were significantly more frequent in the right than in the left lung, as was also the case among 47 matched control patients who had coccidioidomycosis but not meningitis. Of 17 patients whose meningeal symptoms developed during the period studied, all died within 31 months. However, four of eight patients whose onset of symptoms preceded the study period survived for 55-146 months. Patients whose extrapulmonary dissemination involved only the meninges survived significantly longer than did those with more extensive infections. Even without therapy, the white blood cell count in cerebrospinal fluid markedly decreased during the course of the infection. The quantitative analysis provided herein offers a basis for meaningful comparisons with patients enrolled in current and future trials of therapy for coccidioidal meningitis.
- ESPINELINGROFF, A., KISH, C. W., KERKERING, T. M., FROMTLING, R. A., BARTIZAL, K., GALGIANI, J. N., VILLAREAL, K., PFALLER, M. A., GERARDEN, T., RINALDI, M. G., & FOTHERGILL, A. (1992). COLLABORATIVE COMPARISON OF BROTH MACRODILUTION AND MICRODILUTION ANTIFUNGAL SUSCEPTIBILITY TESTS. JOURNAL OF CLINICAL MICROBIOLOGY, 30(12), 3138-3145.
- Calhoun, D. L., White, M. P., Bonner, J. R., Mulholland, J. H., Rumans, L. W., Stevens, D. A., Galgiani, J. N., & Waskin, H. A. (1991). Treatment of systemic sporotrichosis with ketoconazole.. Reviews of infectious diseases, 13(1), 47-51. doi:10.1093/clinids/13.1.47More infoInfections of deep soft tissues with the dimorphic fungus Sporothrix schenckii are uncommon in humans, and therapy has often required toxic drugs. We report our experience in treating 11 patients who had deep-seated sporotrichosis with ketoconazole, a well-tolerated, orally absorbed antifungal agent. Eight infections involved one or more joints, and three involved thoracic, cervical, and widespread cutaneous sites, respectively. For eight patients all evidence of infection resolved during therapy. Sustained remissions (6 months to 5 years) were noted for six patients after the discontinuation of all therapy and for an additional patient 4 years after the initiation of ketoconazole treatment. Durable responses were associated with prolonged treatment with 400-800 mg of ketoconazole daily. Our favorable experience suggests that oral therapy with ketoconazole may benefit other patients with systemic sporotrichosis.
- FISH, D. G., AMPEL, N. M., GALGIANI, J. N., DOLS, C. L., KELLY, P. C., JOHNSON, C. H., PAPPAGIANIS, D., EDWARDS, J. E., WASSERMAN, R. B., CLARK, R. J., ANTONISKIS, D., LARSEN, R. A., ENGLENDER, S. J., & PETERSEN, E. A. (1990). COCCIDIOIDOMYCOSIS DURING HUMAN-IMMUNODEFICIENCY-VIRUS INFECTION - A REVIEW OF 77 PATIENTS. MEDICINE, 69(6), 384-391.
- Galgiani, J. N., Sun, S. H., Clemons, K. V., & Stevens, D. A. (1990). Activity of cilofungin against Coccidioides immitis: differential in vitro effects on mycelia and spherules correlated with in vivo studies.. The Journal of infectious diseases, 162(4), 944-8. doi:10.1093/infdis/162.4.944More infoCilofungin, a new antifungal drug, was found to inhibit mycelial growth of Coccidioides immitis. Light and electron microscopic observations indicated delay in development of the outer hyphal wall. Cilofungin also blocked incorporation of the chitin substrate, N-acetylglucosamine, into mycelia. However, when C. immitis was grown under conditions that induced spherule development, drug effects were dramatically decreased. Furthermore, efficacy of cilofungin in treatment of murine coccidioidomycosis could not be demonstrated. These studies indicate that glucan-synthase inhibitors have activity against C. immitis, and other compounds with different pharmacologic properties or in combination with other antifungal drugs may exploit this biologic effect.
- PFALLER, M. A., RINALDI, M. G., GALGIANI, J. N., BARTLETT, M. S., BODY, B. A., ESPINELINGROFF, A., FROMTLING, R. A., HALL, G. S., HUGHES, C. E., ODDS, F. C., & SUGAR, A. M. (1990). COLLABORATIVE INVESTIGATION OF VARIABLES IN SUSCEPTIBILITY TESTING OF YEASTS. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 34(9), 1648-1654.
- Tucker, R. M., Galgiani, J. N., Denning, D. W., Hanson, L. H., Graybill, J. R., Sharkey, K., Eckman, M. R., Salemi, C., Klein, R. A., Stevens, D. A., & Libke, R. D. (1990). Treatment of coccidioidal meningitis with fluconazole.. Reviews of infectious diseases, 12 Suppl 3(Supplement_3), S380-9. doi:10.1093/clinids/12.supplement_3.s380More infoFluconazole was administered at doses of 50-400 mg/d to 18 patients (15 men, three women) with coccidioidal meningitis. After a mean duration of treatment of 9.8 months, 10 (67%) of 15 assessable patients had responded, one (7%) of 15 had partially responded, and four (27%) of 15 had not responded to therapy. Five (63%) of eight assessable patients receiving fluconazole as sole therapy responded or partially responded. Two patients discontinued fluconazole after initially responding to therapy, and both experienced relapse. The toxicity of fluconazole remains minimal at doses to 400 mg/d. The penetration of fluconazole into cerebrospinal fluid is substantial at all doses studied. Thus fluconazole continues to show promise even as sole therapy against coccidioidal meningitis. Not all patients respond, however, and relapse may be a problem with the currently studied doses and durations of therapy.
- BRONNIMANN, D. A., ADAM, R. D., GALGIANI, J. N., HABIB, M. P., PETERSEN, E. A., PORTER, B., & BLOOM, J. W. (1987). COCCIDIOIDOMYCOSIS IN THE ACQUIRED-IMMUNODEFICIENCY-SYNDROME. ANNALS OF INTERNAL MEDICINE, 106(3), 372-379.
- ROGERS, T. E., & GALGIANI, J. N. (1986). ACTIVITY OF FLUCONAZOLE (UK-49,858) AND KETOCONAZOLE AGAINST CANDIDA-ALBICANS INVITRO AND INVIVO. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 30(3), 418-422.
- Rogers, T. E., & Galgiani, J. N. (1986). Activity of fluconazole (UK 49,858) and ketoconazole against Candida albicans in vitro and in vivo.. Antimicrobial agents and chemotherapy, 30(3), 418-22. doi:10.1128/aac.30.3.418More infoFluconazole (UK 49,858), a new orally administered bis-triazole, was compared with ketoconazole for activity in synthetic broth dilution susceptibility tests against Candida albicans and also in treatment of experimental systemic candidal infections in rats. In vitro studies indicated that fluconazole activity is less sensitive to acidic medium than is that of ketoconazole. At physiologic pH, fluconazole was approximately 16-fold less active than ketoconazole against 35 representative isolates of C. albicans. Two additional isolates (K-1 and K-3) recovered from patients who had failed ketoconazole therapy were 32- to 64-fold more resistant than the median of each drug for other isolates. In animal studies, fluconazole was very effective in prolonging survival of rats infected with a representative candidal strain. With an inoculum sufficient to kill 29 of 38 sham-treated animals, only 1 of 18 animals treated with 0.5 mg of fluconazole per kg per day died compared with 13 of 20 animals treated with 10.0 mg of ketoconazole per kg per day. However, when similar fluconazole treatment was administered to rats infected with the more resistant strain, K-1, no prolongation of survival was found. Thus, in vivo and in vitro results between strains correlated well for fluconazole. However, in comparing results between drugs, ketoconazole was 16-fold more active in vitro and fluconazole was 20-fold more active in vivo. This discrepancy may be due to drug distribution, modes of drug metabolism, or other pharmacologic differences between the two agents.
- Calhoun, D. L., & Galgiani, J. N. (1984). Analysis of pH and buffer effects on flucytosine activity in broth dilution susceptibility testing of Candida albicans in two synthetic media.. Antimicrobial agents and chemotherapy, 26(3), 364-7. doi:10.1128/aac.26.3.364More infoWe examined the influences of different pH levels and three different buffers on flucytosine activity against 12 isolates of Candida albicans in two synthetic media, yeast nitrogen base (YNB) and synthetic amino acid medium-fungal (SAAMF), using broth dilution techniques and measuring the endpoints of visual MICs and turbidimetric 50% inhibitory concentrations. The two media were originally prepared as follows: YNB, unbuffered, pH 5.6; SAAMF, buffered with morpholinepropanesulfonic acid-Tris, pH 7.4; the resultant geometric mean MIC and 50% inhibitory concentration of 5-FC were 78- and 32-fold higher, respectively, in SAAMF. Raising the pH of YNB or lowering the pH of SAAMF had virtually no effect on these differences in MIC and 50% inhibitory concentration in the two media. In contrast, virtually all of the discrepancy appeared to be due to morpholinepropanesulfonic acid-Tris, which exerted concentration-dependent inhibition of flucytosine activity not evident when N-2-hydroxyethylpiperazine-N'-ethanesulfonic acid or phosphate buffer systems were substituted. In other turbidimetric studies, growth was slowed more than 50% in YNB as the pH was raised to 7.4, regardless of which buffer was used. Based on our studies, we recommend modifying the composition of SAAMF by substituting a nonantagonistic buffer if any buffer is to be used with SAAMF in the testing of flucytosine. With this modification, SAAMF warrants further study as a generally applicable medium for fungal-susceptibility testing.
- Galgiani, J. N. (1984). Why not standardize antifungal susceptibility testing. Antimicrobic Newsletter, 1(5), 40. doi:10.1016/0738-1751(84)90017-0
- PONT, A., GRAYBILL JR, ., CRAVEN, P. C., GALGIANI, J. N., DISMUKES, W. E., REITZ, R. E., & STEVENS, D. A. (1984). HIGH-DOSE KETOCONAZOLE THERAPY AND ADRENAL AND TESTICULAR FUNCTION IN HUMANS. ARCHIVES OF INTERNAL MEDICINE, 144(11), 2150-2153.
- BRASS, C., GALGIANI, J. N., BLASCHKE, T. F., DEFELICE, R., OREILLY, R. A., & STEVENS, D. A. (1982). DISPOSITION OF KETOCONAZOLE, AN ORAL ANTIFUNGAL, IN HUMANS. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 21(1), 151-158.
- Defelice, R., Wieden, M. A., & Galgiani, J. N. (1982). The incidence and implications of coccidioidouria.. The American review of respiratory disease, 125(1), 49-52. doi:10.1164/arrd.1918.104.22.168More infoTo estimate the frequency and significance of isolating Coccidioides immitis from urine specimens, we reviewed the records of 75 patients with various forms of coccidioidomycosis of whom 29 had urine cultures for fungi. The 7 patients (24%) who had coccidioidouria appeared similar as a group to those whose cultured urine did not yield C. immitis. Over half of the patients had no other evidence of extrathoracic spread, and in 2 patients with coccidioidouria, illness resolved without treatment. Usually only a few colonies of the fungus were isolated from 200-ml specimens. Thus, our use of a concentrating technique and repeated sampling may account for the high incidence of coccidioidouria in this study. Isolating C. immitis from the urine may be helpful for diagnosis and should lead to further evaluation of the urinary tract for destructive parenchymal lesions. However, as an isolated finding, coccidioidouria does not always indicate the need for therapy.
- DEFELICE, R., JOHNSON, D. G., & GALGIANI, J. N. (1981). GYNECOMASTIA WITH KETOCONAZOLE. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 19(6), 1073-1074.
- Brass, C., Galgiani, J. N., Campbell, S. C., & Stevens, D. A. (1980). Therapy of disseminated or pulmonary coccidioidomycosis with ketoconazole.. Reviews of infectious diseases, 2(4), 656-60. doi:10.1093/clinids/2.4.656More infoOral therapy with ketoconazole for active disseminated or progressive pulmonary coccidioidomycosis was evaluated according to defined criteria of objective improvement for 39 patients, most of whom had received other antifungal chemotherapy. Minimal inhibitory concentrations of ketoconazole for isolates of Coccidioides immitis were below mean peak serum concentrations. Eighteen patients responded at all sites of disease, one patient failed to respond, and the others either are being evaluated or cannot be evaluated. Most patients who responded to therapy required more than three months of treatment before the response was clearly noted. Responses were seen with skin, soft tissue, skeletal, and pulmonary infection as well as other conditions. Reversions of cultures for C. immitis to negative and decreases in titers of complement-fixing antibody were common. One patient relapsed after a course of therapy of only four months. In general, patients with skin disease who responded required only 200 mg per day, whereas those with skeletal disease required 400 mg per day. Adverse effects were uncommon despite extensive monitoring and were generally limited to transient nausea (with vomiting in patients receiving 400 mg per day). The data show that ketoconazole appears to be a promising new drug for treatment of coccidioidomycosis.
- Carnevale, N. T., Stevens, D. A., Langston, J. W., Herrick, M. K., & Galgiani, J. N. (1980). Amphotericin B-induced myelopathy.. JAMA Internal Medicine, 140(9), 1189-1192. doi:10.1001/archinte.1980.00330200065022More info• Two patients with coccidioidal meningitis experienced transient neurologic deficits shortly after receiving intrathecal injections of amphotericin B. Continuation of treatment eventually led to a severe flaccid paraparesis with a thoracic sensory level in one patient, and a partial Brown-Sequard's syndrome in the other. Myelography was normal in both, with no evidence of arachnoiditis. Autopsy findings in the first patient showed a focal area of necrosis in the left half of the spinal cord consistent with the patient's clinical findings during life. The distribution of the lesion corresponded to the area supplied by a central sulcal artery. Amphotericin B may exert a direct toxic effect on the spinal cord or its vascular supply when given intrathecally. (Arch Intern Med 140:1189-1192, 1980)
- Galgiant, J. N., Stevens, D. A., & Galgiani, J. N. (1978). Turbidimetric studies of growth inhibition of yeasts with three drugs: inquiry into inoculum-dependent susceptibility testing, time of onset of drug effect, and implications for current and newer methods.. Antimicrobial agents and chemotherapy, 13(2), 249-54. doi:10.1128/aac.13.2.249More infoSusceptibility testing with the broth-dilution visual end-point method is inoculum dependent with miconazole and 5-fluorocystosine, but not with amphotericin B. Turbidimetric measurements of yeast growth in the presence of antifungal drugs were therefore performed. With miconazole and 5-fluorocytosine, over the range of concentrations studied, growth occurred until a plateau phase was reached. With drug present prior to log phase growth, a delayed onset of effect was noted which was proportional to the generation time of the organism. With amphotericin B, in contrast, there was sharp transition with increasing drug concentration from no inhibition to complete arrest of growth, and no relation of onset of effect to generation time. These findings provide a possible explanation of inoculum dependence; i.e., at higher inocula, partially inhibited but growing yeasts become visible at higher drug concentrations. Supporting evidence derives from observations with different starting inocula, varying time of reading end points, and other methods of studying growth inhibition. The delay in miconazole and 5-fluorocytosine effect described suggests that rapid methods of susceptibility testing should be utilized with cultures already in log phase.
- Carnevale, N. T., Galgiani, J. N., Stevens, D. A., Langston, J. W., & Herrick, M. K. (1977). Amphotericin B-Induced Myelopathy. Neurology, 27(4).More infoone patient, and a partial Brown-S\l=e'\quard'ssyndrome in the other. Myelography was normal in both, with no evidence of arachnoiditis. Autopsy findings in the first patient showed a focal area of necrosis in the left half of the spinal cord consistent with the patient's clinical findings during life. The distribution of the lesion corresponded to the area supplied by a central sulcal artery. Amphotericin B may exert a direct toxic effect on the spinal cord or its vascular supply when given intrathecally. (Arch Intern Med 140:1189-1192, 1980) CJymptoms suggesting spinal cord or radicular dysfunc^ tion often occur in patients undergoing treatment with intrathecally administered amphotericin B. These compli¬ cations are usually attributed to arachnoiditis near or at the site of injection.16 The purpose of this report is to present two cases in which clinical, myelographie, and (in one) pathologic evidence failed to support the presence of an amphotericin B-induced arachnoiditis, but rather sug¬ gested direct damage to the spinal cord. This potentially catastrophic complication of intrathecally administered amphotericin may be avoidable, since transient symp¬ toms and signs appear to precede permanent neurologic deficits.
- GALGIANI, J. N., & STEVENS, D. A. (1976). ANTIMICROBIAL SUSCEPTIBILITY TESTING OF YEASTS - TURBIDIMETRIC TECHNIQUE INDEPENDENT OF INOCULUM SIZE. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 10(4), 721-726.
- LUTWICK, L. I., GALGIANI, J. N., JOHNSON, R. H., & STEVENS, D. A. (1976). VISCERAL FUNGAL INFECTIONS DUE TO PETRIELLIDIUM BOYDII (ALLESCHERIA BOYDII) - INVITRO DRUG SENSITIVITY STUDIES. AMERICAN JOURNAL OF MEDICINE, 61(5), 632-640.
- Lutwick, L. I., Galgiani, J. N., Johnson, R. H., & Stevens, D. A. (1976). Visceral fungal infections due to Petriellidium boydii (allescheria boydii). In vitro drug sensitivity studies.. The American journal of medicine, 61(5), 632-40. doi:10.1016/0002-9343(76)90141-8More infoFour patients with visceral infections due to the fungus Petriellidium boydii, who were recently hospitalized in our institutions, are described. Three of the patients were compromised hosts; in the fourth patient, infection occurred after trauma. All had received prior steroid and antibiotic therapy. Studies of patients with mycetoma or secondary infection of a pulmonary cavity due to this organism and of patients with visceral infections are reviewed. Because of histologic similarities to Aspergillus species, infections due to P. boydii may have been misdiagnosed in the past if the infecting fungus was not isolated in culture. The fungus has been shown to be resistant in vitro to currently available antifungal agents. Resistance to amphotericin and 5-fluorocytosine is demonstrated in our studies. There are few reports of successful chemotherapy of any manifestation of this infection, and no such reports of visceral disease. We demonstrate in vitro sensitivity of isolates in our cases and in others to micronazole, a new antimicrobial agent; this drug may be indicated for treatment of disease due to P. boydii.
- Donovan, F., Ramadan, F., Ellingson, K., & Galgiani, J. N. (2020, March). A Clinical Profile of Newly Diagnosed Valley Fever in Hospital Settings. SHEA/CDC Decennial 6th International Conference on Healthcare Associated Infections. Atlanta, GA: SHEA/CDC Decennial 6th International Conference on Healthcare Associated Infections.
- Galgiani, J. N. (2019, April). Coccidioidomycosis (Valley fever): A work-related disease. American Occupational Health Conference. Anaheim CA: American College of Occupational and Environmental Medicine.
- Galgiani, J. N. (2019, April). Development of Nikkomycin Z in an Academic Setting. 2nd Antifungal Drug Discovery Symposium. Durham NC: Duke University.
- Galgiani, J. N. (2019, March). Vaccines to Prevent Coccidioidomycosis: 60 Years and Counting. Vaccine Strategies for Endemic Fungal Pathogens. Rockville MD: NIH-NIAID.
- Galgiani, J. N., Leroy, G. A., Donovan, F., Gu, Y., & Song, H. (2021, November). Automated Biomedical Lexicon Creation with Graph Search for Word Embeddings. IEEE CHASE. Washington DC.
- Leroy, G. A., Galgiani, J. N., Donovan, F., Pu, J., Ramadan, F., & Ellingson, K. (2019, November). Valley Fever Phenotypic Presentation: Exploration of a Precision Medicine Approach to Clinical Decision Support". Reimagine Health: Is my fate in my genes?.
- Lussier, Y. A., Galgiani, J. N., Frelinger, J. A., Holland, S. M., Shubitz, L., Vitali, F., Pouladi, N., Chaput, A. L., Powell, D. P., Hsu, A. P., Stewart, E., Waldman, S., Bays, D., Thompson, G. R., & Berghout, J. (2019, Nov). Collaborative Arizona and California combined cohort to investigate the role of rare genetic variants in susceptibility to disseminated coccidioidomycosis (Valley fever). reimagine Health: Is my fate in my genes? 2nd Annual University of Arizona Research Symposium. Phoenix, AZ: University of Arizona.
- Lussier, Y. A., Galgiani, J. N., Frelinger, J. A., Holland, S. M., Shubitz, L., Vitali, F., Pouladi, N., Chaput, A. L., Powell, D. P., Hsu, A. P., Stewart, E., Waldman, S., Bays, D., Thompson, G. R., & Berghout, J. (2019, Oct). Identification of rare variants contributing to disseminated coccidioidomycosis after infection. American Society of Human Genetics. Houston, TX.More infoASHG Abstract ID 30302128
- Butkiewicz, C. D., Lewis, M. L., Trinh, H. T., Galgiani, J. N., Shubitz, L., Frelinger, J. A., & Orbach, M. J. (2017, August). Further characterization of the avirulent delta-cps1 Coccidioides vaccine in mice. Seventh International Coccidioidomycosis Symposium. Stanford University, Stanford, CA: Stanford Center for Continuing Medical Education.More infoPoster presentation on further analysis of the duration and protection of the delta-cps1 valley fever vaccine.
- Frelinger, J. A., Galgiani, J. N., Orbach, M. J., D, B. C., Trinh, H. T., Lewis, M. L., Shubitz, L., & Powell, D. A. (2017, August). Determining Mechanisms of Protection in a Live Attenuated Coccidiodes Vaccine. 7th International Coccidioidomycosis Symposium. Stanford University, Stanford, CA: Stanford Center for Continuing Medical Education.More infoposter presentation describing the immune response of mice to our valley fever vaccine.
- Lussier, Y. A., Galgiani, J. N., Frelinger, J. A., Holland, S. M., Hsu, A., Powell, D., Li, H., Li, Q., & Berghout, J. (2017, August). SINGLE-SUBJECT TRANSCRIPTOME PROFILING OF STAT4 MUTANT PATIENT PBMCS SUGGESTS ALTERED RESPONSIVENESS TO COCCI LYSATE STIMULATION. PROCEEDINGS OF THE COCCIDIOIDOMYCOSIS STUDY GROUP 61st ANNUAL MEETING in collaboration with the 7th INTERNATIONAL COCCIDIOIDOMYCOSIS SYMPOSIUM. Stanford University.
- Orbach, M. J., Galgiani, J. N., Frelinger, J. A., Buntzman, A. S., Trinh, H., Lewis, L., Shubitz, L., & Mandel, M. A. (2017, March, 2017). Construction and Efficacy of delta-cps1, a live attenuated vaccine for coccidioidomycosis. 29th Fungal Genetics Conference. Asilomar Conference Center, Pacific Grove, CA: Genetics Society of America.More infoA poster was presented, and was selected for a poster talk in the Concurrent Session "Human Pathogenic Fungi", which was presented by Dr. Mandel.The abstracts are published as a supplement of Fungal Genetics Reports.
- Orbach, M. J., Galgiani, J. N., Frelinger, J. A., Buntzman, A., Lewis, M. L., Trinh, H., Shubitz, L., & Mandel, M. A. (2017, April 1). Construction and efficacy of delta-cps1, a live attenuated vaccine for coccidioidomycosis. 56th Annual ASM Regional Meeting- Southwest Region. University of Arizona: American Society of Microbiology.