Michael O Daines

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Scholarly Contributions

Chapters

• Daines, M. O., & Phan, H. (2018). Allergic Rhinitis. In Pharmacotherapy: Principles and Practice. 5th Ed.(pp 967-982). McGraw-Hill.
• Daines, M., Daines, M. O., & Morgan, W. J. (2013). The emergence of monoclonal antibodies. In Landmark Papers in Allergy. Oxford University Press. doi:10.1093/MED/9780199651559.003.0077

Journals/Publications

• Covar, R., Lazarus, S. C., Krishnan, J. A., Blake, K. V., Sorkness, C. A., Dyer, A. M., Lang, J. E., Lugogo, N. L., Mauger, D. T., Wechsler, M. E., Wenzel, S. E., Cardet, J. C., Castro, M., Israel, E., Phipatanakul, W., King, T. S., & , N. H. (2024). Association of Sputum Eosinophilia With Easily Measured Type-2 Inflammatory Biomarkers in Untreated Mild Persistent Asthma. The journal of allergy and clinical immunology. In practice, 12(4), 960-969.e6.
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  A multicenter clinical trial in patients with mild persistent asthma indicated that response to inhaled corticosteroids (ICS) is limited to those with sputum eosinophilia. However, testing for sputum eosinophilia is impractical in most clinical settings.
• Eremija, J., & Daines, M. O. (2023).

  Neuropsychiatric Testing Provides Objective Insight Into Beneficial Effects Of Intravenous Immunoglobulins In Patients With Pediatric Acute-Onset Neuropsychiatric Syndrome

  . JACI, Abstract for meeting. doi:10.1016/j.jaci.2022.12.253
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  Pediatric Acute-Onset Neuropsychiatric Syndrome (PANS) is defined by acute beginning of diverse neuropsychiatric symptoms, likely in the setting of underlying immune dysfunction. Thus, immunomodulatory interventions have a crucial therapeutic role, but objective post-treatment evaluations remain challenging given disease complexity. We used standardized neuropsychiatric testing to assess how intravenous immunoglobulins (IVIG) impact cognitive function.
• Eremija, J., Patel, S., Rice, S., & Daines, M. (2023). Intravenous immunoglobulin treatment improves multiple neuropsychiatric outcomes in patients with pediatric acute-onset neuropsychiatric syndrome. Frontiers in pediatrics, 11, 1229150.
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  Pediatric Acute-Onset Neuropsychiatric Syndrome (PANS) is defined by acute onset of diverse neuropsychiatric manifestations, presumably in the setting of underlying immune dysfunction We used standardized neuropsychological testing to assess how intravenous immunoglobulins (IVIG) impact neurological and cognitive functions in PANS patients by comparing pretreatment with post-treatment scores. A 5-year retrospective study was undertaken in Children's Postinfectious Autoimmune Encephalopathy Center at University of Arizona. We identified 12 children diagnosed with PANS and treated with immunomodulatory IVIG doses, who also completed neuropsychological testing before and after treatment. We tracked multiple patient characteristics, type/timeline of testing, and number of IVIG courses. Score change of 1 standard deviation in any tested domain/subdomain was considered improvement. We further reviewed records for laboratory signs of triggering infection and immune dysfunction. Improvement occurred in 11/12 patients, in one or multiple domains/subdomains, independently of time between disease onset and IVIG initiation (0-7 years). Participants received 1-7 IVIG courses. Improvement was primarily seen in memory (58%), sensory-motor (37%) and visual-motor integration (30%). In 5/12 patients we detected hypogammaglobulinemia requiring ongoing IVIG replacement, one patient had isolated low IgA. Only one patient had to discontinue IVIG therapy due to severe adverse effects. Standardized neuropsychological testing represents an important tool to objectively measure improvement in PANS patients. IVIG was tolerated well and showed efficacy in the vast majority of participants, independently from timelapse since disease onset, emphasizing impact of immunomodulation in PANS. Significant presence of baseline hypogammaglobulinemia in children with PANS emphasizes the presumed role of immune dysfunction in disease pathogenesis.
• Eremija, J., Patel, S., Rice, S., & Daines, M. O. (2023). Intravenous immunoglobulin treatment improves multiple neuropsychiatric outcomes in patients with pediatric acute-onset neuropsychiatric syndrome. Front Pediatri. doi:10.3389/fped.2023.1229150
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  Pediatric Acute-Onset Neuropsychiatric Syndrome (PANS) is defined by acute onset of diverse neuropsychiatric manifestations, presumably in the setting of underlying immune dysfunction . We used standardized neuropsychological testing to assess how intravenous immunoglobulins (IVIG) impact neurological and cognitive functions in PANS patients by comparing pretreatment with post-treatment scores. A 5-year retrospective study was undertaken in Children's Postinfectious Autoimmune Encephalopathy Center at University of Arizona. We identified 12 children diagnosed with PANS and treated with immunomodulatory IVIG doses, who also completed neuropsychological testing before and after treatment. We tracked multiple patient characteristics, type/timeline of testing, and number of IVIG courses. Score change of 1 standard deviation in any tested domain/subdomain was considered improvement. We further reviewed records for laboratory signs of triggering infection and immune dysfunction. Improvement occurred in 11/12 patients, in one or multiple domains/subdomains, independently of time between disease onset and IVIG initiation (0–7 years). Participants received 1–7 IVIG courses. Improvement was primarily seen in memory (58%), sensory-motor (37%) and visual-motor integration (30%). In 5/12 patients we detected hypogammaglobulinemia requiring ongoing IVIG replacement, one patient had isolated low IgA. Only one patient had to discontinue IVIG therapy due to severe adverse effects. Standardized neuropsychological testing represents an important tool to objectively measure improvement in PANS patients. IVIG was tolerated well and showed efficacy in the vast majority of participants, independently from timelapse since disease onset, emphasizing impact of immunomodulation in PANS. Significant presence of baseline hypogammaglobulinemia in children with PANS emphasizes the presumed role of immune dysfunction in disease pathogenesis.
• Booth, N. A., Freeman, C. M., Wright, B. L., Rukasin, C., Badia, P., Daines, M., Bauer, C. S., & Miller, H. (2022). Severe Combined Immunodeficiency (SCID) Screening in Arizona: Lessons Learned from the First 2 Years. Journal of clinical immunology, 42(6), 1321-1329.
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  The incidence of severe combined immunodeficiency (SCID) in the USA was reported as 1 in 58,000 live births. In Arizona, it was anticipated that newborn screening would identify two to four cases of SCID per year. This estimate did not consider ethnic nuances in Arizona, with higher percentages of Native American and Hispanic populations compared to national percentages. The true incidence of SCID and non-SCID T cell lymphopenia has not previously been reported in Arizona.
• Bacharier, L. B., Maspero, J. F., Katelaris, C. H., Fiocchi, A. G., Gagnon, R., de Mir, I., Jain, N., Sher, L. D., Mao, X., Liu, D., Zhang, Y., Khan, A. H., Kapoor, U., Khokhar, F. A., Rowe, P. J., Deniz, Y., Ruddy, M., Laws, E., Patel, N., , Weinreich, D. M., et al. (2021). Dupilumab in Children with Uncontrolled Moderate-to-Severe Asthma. The New England journal of medicine, 385(24), 2230-2240.
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  Children with moderate-to-severe asthma continue to have disease complications despite the receipt of standard-of-care therapy. The monoclonal antibody dupilumab has been approved for the treatment of adults and adolescents with asthma as well as with other type 2 inflammatory diseases.
• Daines, M. O., Zhu, l., Pereira, R., Zhou, X., Bondy, C., Pryor, B. M., Zhou, J., & Chen, Y. (2021). Alternaria induces airway epithelial cytokine expression independent of protease-activated receptor. Respirology, 25(5), 502-510. doi:10.1111/resp.13675
• Daines, M., Pereira, R., Cunningham, A., Pryor, B., Besselsen, D. G., Liu, Y., Luo, Q., & Chen, Y. (2021). Novel Mouse Models of Fungal Asthma. Frontiers in cellular and infection microbiology, 11, 683194.
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  is a ubiquitous fungus and a major allergen associated with the development of asthma. Inhalation of intact spores is the primary cause of human exposure to fungal allergen. However, allergen-rich cultured fungal filtrates are oftentimes used in the current models of fungal sensitization that do not fully reflect real-life exposures. Thus, establishing novel spore exposure models is imperative. In this study, we established novel fungal exposure models of both adult and neonate to live spores. We examined pathophysiological changes in the spore models as compared to the non-exposure controls and also to the conventional filtrate models. While both filtrate- and spore-exposed adult BALB/c mice developed elevated airway hyperresponsiveness (AHR), filtrates induced a greater IgE mediated response and higher broncholavage eosinophils than spores. In contrast, the mice exposed to spores had higher numbers of neutrophils. Both exposures induced comparable levels of lung tissue inflammation and mucous cell metaplasia (MCM). In the neonatal model, exposure to spores resulted in a significant increase of AHR in both adult and neonatal mice. Increased levels of IgE in both neonatal and adult mice exposed to spores was associated with increased eosinophilia in the treatment groups. Adult demonstrated increased numbers of lymphocytes that was paralleled by increased IgG1 production. Both adults and neonates demonstrated similarly increased eosinophilia, IgE, tissue inflammation and MCM.
• Rice, S. A., Cabrera, A., Daines, M. O., Spiekerman, C., & Klinger, P. (2021). A Case Study of Acute Onset Obsessive-Compulsive Symptoms in a Pediatric Patient. Journal of Multidisciplinary Clinical Case Reports, 1a(103), 1-6. doi:doi: https://doi.org/10.33790/ jmccr1100103
• Daines, M., Zhu, L., Pereira, R., Zhou, X., Bondy, C., Pryor, B. M., Zhou, J., & Chen, Y. (2020). Alternaria induces airway epithelial cytokine expression independent of protease-activated receptor. Respirology (Carlton, Vic.).
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  A novel fungal allergen, Alternaria (Alt), has been previously shown to associate with the pathogenesis of allergic rhinitis and bronchial asthma, particularly in arid and semi-arid regions. Airway epithelial cells are among the first to encounter Alt, and epithelial cytokine production and subsequent airway inflammation are early events in the response to Alt exposure. However, the underlying mechanism is unclear. As protease-activated receptor 2 (PAR2) has been implicated in most of the Alt-induced biological events, we investigated the regulation of airway inflammation and epithelial cytokine expression by PAR2.
• Fitzpatrick, A. M., Bacharier, L. B., Guilbert, T. W., Jackson, D. J., Szefler, S. J., Beigelman, A., Cabana, M. D., Covar, R., Holguin, F., Lemanske, R. F., Martinez, F. D., Morgan, W., Phipatanakul, W., Pongracic, J. A., Zeiger, R. S., Mauger, D. T., & , N. A. (2019). Phenotypes of Recurrent Wheezing in Preschool Children: Identification by Latent Class Analysis and Utility in Prediction of Future Exacerbation. The journal of allergy and clinical immunology. In practice, 7(3), 915-924.e7.
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  Recurrent preschool wheezing is a heterogeneous disorder with significant morbidity, yet little is known about phenotypic determinants and their impact on clinical outcomes.
• Guilbert, T. W., Bacharier, L. B., Mauger, D. T., Phipatanakul, W., Szefler, S. J., Boehmer, S., Beigelman, A., Fitzpatrick, A. M., Jackson, D. J., Baxi, S. N., Benson, M., Burnham, C. D., Cabana, M. D., Castro, M., Chmiel, J. F., Covar, R., Daines, M., Gaffin, J. M., Gentile, D. A., , Holguin, F., et al. (2019). Challenges in assessing the efficacy of systemic corticosteroids for severe wheezing episodes in preschool children. The Journal of allergy and clinical immunology, 143(5), 1934-1937.e4.
• Lazarus, S. C., Krishnan, J. A., King, T. S., Lang, J. E., Blake, K. V., Covar, R., Lugogo, N., Wenzel, S., Chinchilli, V. M., Mauger, D. T., Dyer, A. M., Boushey, H. A., Fahy, J. V., Woodruff, P. G., Bacharier, L. B., Cabana, M. D., Cardet, J. C., Castro, M., Chmiel, J., , Denlinger, L., et al. (2019). Mometasone or Tiotropium in Mild Asthma with a Low Sputum Eosinophil Level. The New England journal of medicine, 380(21), 2009-2019.
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  In many patients with mild, persistent asthma, the percentage of eosinophils in sputum is less than 2% (low eosinophil level). The appropriate treatment for these patients is unknown.
• Zhou, J., Zhou, X., Daines, M., Zhu, L., Pereira, R., Bondy, C., Pryor, B. M., & Chen, Y. (2019). Alternaria induces airway epithelial cytokine expression independent of protease‐activated receptor. Respirology, 25(5), 502-510. doi:10.1111/resp.13675
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  A novel fungal allergen, Alternaria (Alt), has been previously shown to associate with the pathogenesis of allergic rhinitis and bronchial asthma, particularly in arid and semi-arid regions. Airway epithelial cells are among the first to encounter Alt, and epithelial cytokine production and subsequent airway inflammation are early events in the response to Alt exposure. However, the underlying mechanism is unclear. As protease-activated receptor 2 (PAR2) has been implicated in most of the Alt-induced biological events, we investigated the regulation of airway inflammation and epithelial cytokine expression by PAR2.Wild-type (WT) and Par2 knockout (Par2-KO) mice were used to evaluate the in vivo role of PAR2. Primary human and mouse airway epithelial cells were used to examine the mechanistic basis of epithelial cytokine regulation in vitro.Surprisingly, Par2 deficiency had no negative impact on the change of lung function, inflammation and cytokine production in the mouse model of Alt-induced asthma. Alt-induced cytokine production in murine airway epithelial cells from Par2-KO mice was not significantly different from the WT cells. Consistently, PAR2 knockdown in human cells also had no effect on cytokine expression. In contrast, the cytokine expressions induced by synthetic PAR2 agonist or other asthma-related allergens (e.g. cockroach extracts) were indeed mediated via a PAR2-dependent mechanism. Finally, we found that EGFR pathway was responsible for Alt-induced epithelial cytokine expression.The activation of EGFR, but not PAR2, was likely to drive the airway inflammation and epithelial cytokine production induced by Alt.
• Hiranrattana, A., Stern, D. A., Guerra, S., Halonen, M., Wright, A. L., Daines, M., Martinez, F. D., & Morgan, W. J. (2018). sensitisation at age 6 years is associated with subsequent airway hyper-responsiveness in non-asthmatics. Thorax, 73(12), 1170-1173.
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  In the non-selected birth cohort Tucson Children's Respiratory Study, early sensitisation to was associated with increased airway hyper-responsiveness (AHR) into adult life among non-asthmatics. The increase in AHR was of a similar magnitude to that seen for sensitised asthmatics and was primarily evident among those who were overweight or obese. In contrast, there was no significant association between early sensitisation to aeroallergens other than and AHR among non-asthmatics. Why this group of sensitised individuals without asthma demonstrated increased AHR of a magnitude similar to asthmatics is unknown and requires further investigation.
• Jackson, D. J., Bacharier, L. B., Mauger, D. T., Boehmer, S., Beigelman, A., Chmiel, J. F., Fitzpatrick, A. M., Gaffin, J. M., Morgan, W. J., Peters, S. P., Phipatanakul, W., Sheehan, W. J., Cabana, M. D., Holguin, F., Martinez, F. D., Pongracic, J. A., Baxi, S. N., Benson, M., Blake, K., , Covar, R., et al. (2018). Quintupling Inhaled Glucocorticoids to Prevent Childhood Asthma Exacerbations. The New England journal of medicine, 378(10), 891-901.
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  Asthma exacerbations occur frequently despite the regular use of asthma-controller therapies, such as inhaled glucocorticoids. Clinicians commonly increase the doses of inhaled glucocorticoids at early signs of loss of asthma control. However, data on the safety and efficacy of this strategy in children are limited.
• Morris, C. R., Mauger, D. T., Suh, J. H., Phipatanakul, W., Sheehan, W. J., Moy, J. N., Paul, I. M., Szefler, S. J., Jackson, D. J., Fitzpatrick, A. M., & , N. I. (2018). Glutathione and arginine levels: Predictors for acetaminophen-associated asthma exacerbation?. The Journal of allergy and clinical immunology, 142(1), 308-311.e9.
• Wright, A. L., Daines, M. O., Morgan, W. J., Martinez, F. D., Halonen, M., Guerra, S., Stern, D. A., & Hiranrattana, A. (2018). Alternaria sensitisation at age 6 years is associated with subsequent airway hyper-responsiveness in non-asthmatics. Thorax. doi:10.1136/thoraxjnl-2017-210325
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  In the non-selected birth cohort Tucson Children’s Respiratory Study, early sensitisation to Alternaria was associated with increased airway hyper-responsiveness (AHR) into adult life among non-asthmatics. The increase in AHR was of a similar magnitude to that seen for Alternaria sensitised asthmatics and was primarily evident among those who were overweight or obese. In contrast, there was no significant association between early sensitisation to aeroallergens other than Alternaria and AHR among non-asthmatics. Why this group of Alternaria sensitised individuals without asthma demonstrated increased AHR of a magnitude similar to asthmatics is unknown and requires further investigation.
• Yee, M. C., Nichols, H. L., Polley, D., Mahmoud, S., Pal, K., Lee, K., Wilson, E. H., Daines, M. O., Hollenberg, M. D., Boitano, S. A., & DeFea, K. A. (2018). Protease-activated Receptor-2 Signaling through beta-Arrestin-2 Mediates Alternaria Alkaline Serine Protease-induced Airway Inflammation. Am J Physiol Lung Cell Mol Physiol. doi:10.1152/ajplung.00196.2018
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  Alternaria alternata is a fungal allergen associated with severe asthma and asthma exacerbations. Similar to other asthma-associated allergens, Alternaria secretes a serine-like trypsin protease(s) that is thought to act through the G protein-coupled receptor, protease-activated receptor-2 (PAR2), to induce asthma symptoms. However, specific mechanisms underlying Alternaria-induced PAR2 activation and signaling remain ill-defined. We sought to determine whether Alternaria-induced PAR2 signaling contributed to asthma symptoms via a PAR2/beta-arrestin signaling axis, to identify the protease activity responsible for PAR2 signaling and determine if protease activity was sufficient for Alternaria-induced asthma symptoms in animal models. We initially used in vitro models to demonstrate Alternaria-induced PAR2/beta-arrestin-2 signaling. Alternaria filtrates were then used to sensitize and challenge wild-type, PAR2(-/-) and beta-arrestin-2(-/-) mice in vivo. Intranasal administration of Alternaria filtrate resulted in a protease-dependent increase of airway inflammation and mucin production in wild-type mice, but not PAR2(-/-) or beta-arrestin-2(-/-) mice. Protease was isolated from Alternaria preparations and select in vitro and in vivo experiments were repeated to evaluate sufficiency of the isolated Alternaria protease to induce asthma phenotype. Administration of a single isolated serine protease from Alternaria, Alternaria Alkaline Serine Protease (AASP), was sufficient to fully activate PAR2 signaling and induce beta-arrestin-2-dependent eosinophil and lymphocyte recruitment in vivo. In conclusion, Alternaria filtrates induce airway inflammation and mucus hyperplasia largely via AASP using the PAR2/beta-arrestin signaling axis. Thus, beta-arrestin-biased PAR2 antagonists represent novel therapeutic targets for treating aeroallergen-induced asthma.
• Yee, M. C., Nichols, H. L., Polley, D., Saifeddine, M., Pal, K., Lee, K., Wilson, E. H., Daines, M. O., Hollenberg, M. D., Boitano, S., & DeFea, K. A. (2018). Protease-activated receptor-2 signaling through β-arrestin-2 mediates Alternaria alkaline serine protease-induced airway inflammation. American journal of physiology. Lung cellular and molecular physiology, 315(6), L1042-L1057.
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  Alternaria alternata is a fungal allergen associated with severe asthma and asthma exacerbations. Similarly to other asthma-associated allergens, Alternaria secretes a serine-like trypsin protease(s) that is thought to act through the G protein-coupled receptor protease-activated receptor-2 (PAR) to induce asthma symptoms. However, specific mechanisms underlying Alternaria-induced PAR activation and signaling remain ill-defined. We sought to determine whether Alternaria-induced PAR signaling contributed to asthma symptoms via a PAR/β-arrestin signaling axis, identify the protease activity responsible for PAR signaling, and determine whether protease activity was sufficient for Alternaria-induced asthma symptoms in animal models. We initially used in vitro models to demonstrate Alternaria-induced PAR/β-arrestin-2 signaling. Alternaria filtrates were then used to sensitize and challenge wild-type, PAR and β-arrestin-2 mice in vivo. Intranasal administration of Alternaria filtrate resulted in a protease-dependent increase of airway inflammation and mucin production in wild-type but not PAR or β-arrestin-2 mice. Protease was isolated from Alternaria preparations, and select in vitro and in vivo experiments were repeated to evaluate sufficiency of the isolated Alternaria protease to induce asthma phenotype. Administration of a single isolated serine protease from Alternaria, Alternaria alkaline serine protease (AASP), was sufficient to fully activate PAR signaling and induce β-arrestin-2-dependent eosinophil and lymphocyte recruitment in vivo. In conclusion, Alternaria filtrates induce airway inflammation and mucus hyperplasia largely via AASP using the PAR/β-arrestin signaling axis. Thus, β-arrestin-biased PAR antagonists represent novel therapeutic targets for treating aeroallergen-induced asthma.
• Yeee, M. C., Nichols, H. L., Polley, D., Pal, K., Lee, K., Daines, M. O., Boitano, S. A., Hollenberg, M. D., & DeFea, K. A. (2017). Protease-activated receptor 2-induced signaling through β-arrestin-2 mediates Alternaria serine protease-induced airway inflammation. PLoS One, In Revision.
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  Protease-Activated Receptor-2 (PAR2) is a G-protein-coupled receptor that is activated by a variety of trypsin-like serine proteases, including those found in common allergens such as Alternaria fungi, which are causative in asthma-associated morbidity and mortality worldwide. PAR2 signals through both G-protein and β-arrestin-dependent pathways and β-arrestin-2 mediates PAR2-induced leukocyte chemotaxis into the airway. Previous studies showed that an unidentified serine protease activity in Alternaria alternata can activate PAR2 to trigger an inflammatory cell pathology in murine lung. We hypothesized that a serine-protease-mediated PAR2/β-arrestin signaling axis plays a key role in Alternaria-induced lung inflammation. Here we isolate, from Alternaria extracts, the single serine protease responsible for PAR2 activation, AASP (Alternaria Alkaline Serine Protease). Both extracts and isolated AASP promote PAR2 β-arrestin-dependent signaling in cultured cells. Intranasal administration of Alternaria extract in wild-type (but not β-arrestin-2-/- or PAR2-/- mice) increases airway epithelial damage, mucin production, and recruitment of eosinophils, CD4+T-cells and macrophages and this response is abolished by administration of soybean trypsin inhibitor (SBTI). Administration of AASP alone is sufficient to induce eosinophil and lymphocyte recruitment. Our data demonstrate that a single serine protease in an aeroallergen like Alternaria alternata can activate PAR2 signaling through β-arrestin-2 to cause an inflammatory asthma phenotype. Thus, β-arrestin-biased PAR2 antagonists represent attractive therapeutic targets for treating aeroallergen-induced asthma.
• Addison, K. J., Morse, J., Robichaud, A., Daines, M. O., & Ledford, J. G. (2017). A Novel System to Test Bronchodilators. Journal of infectious pulmonary diseases, 3(1).
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  The incidence and severity of asthma continue to rise worldwide. β-agonists are the most commonly prescribed therapeutic for asthma management but have less efficacy for some subsets of asthmatic patients and there are concerns surrounding the side effects from their long-term persistent use. The demand to develop novel asthma therapeutics highlights the need for a standardized approach to effectively screen and test potential bronchoprotective compounds using relevant animal models. Here we describe a validated method of testing potential therapeutic compounds for their fast-acting efficacy during the midst of an induced bronchoconstriction in a house dust mite challenged animal model.
• Daines, M. O., Patel, S., Daines, M. O., & Carr, T. F. (2017). ORAI1 Mutation in a Child with Primary Immunodeficiency-9. The Journal of Allergy and Clinical Immunology, 139(2), AB17. doi:10.1016/j.jaci.2016.12.010
• Frankovich, J., Swedo, S., Murphy, T., Dale, R. C., Agalliu, D., Williams, K., Daines, M., Hornig, M., Chugani, H., Sanger, T., Muscal, E., Pasternack, M., Cooperstock, M., Gans, H., Zhang, Y., Cunningham, M., Bernstein, G., Bromberg, R., Willett, T., , Brown, K., et al. (2017). Clinical Management of Pediatric Acute-Onset Neuropsychiatric Syndrome: Part II-Use of Immunomodulatory Therapies. Journal of child and adolescent psychopharmacology, 27(7), 574-593.
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  Pediatric Acute-onset Neuropsychiatric Syndrome (PANS) is a clinically heterogeneous disorder with a number of different etiologies and disease mechanisms. Inflammatory and postinfectious autoimmune presentations of PANS occur frequently, with some clinical series documenting immune abnormalities in 75%-80% of patients. Thus, comprehensive treatment protocols must include immunological interventions, but their use should be reserved only for PANS cases in which the symptoms represent underlying neuroinflammation or postinfectious autoimmunity, as seen in the PANDAS subgroup (Pediatric Autoimmune Neuropsychiatric Disorders associated with Streptococcal infections). The PANS Research Consortium (PRC) immunomodulatory task force is comprised of immunologists, rheumatologists, neurologists, infectious disease experts, general pediatricians, psychiatrists, nurse practitioners, and basic scientists with expertise in neuroimmunology and PANS-related animal models. Preliminary treatment guidelines were created in the Spring of 2014 at the National Institute of Health and refined over the ensuing 2 years over conference calls and a shared web-based document. Seven pediatric mental health practitioners, with expertise in diagnosing and monitoring patients with PANS, were consulted to create categories in disease severity and critically review final recommendations. All authors played a role in creating these guidelines. The views of all authors were incorporated and all authors gave final approval of these guidelines. Separate guidelines were created for the use of immunomodulatory therapies in PANS patients with (1) mild, (2) moderate-to-severe, and (3) extreme/life-threatening severity. For mildly impairing PANS, the most appropriate therapy may be "tincture of time" combined with cognitive behavioral therapy and other supportive therapies. If symptoms persist, nonsteroidal anti-inflammatory drugs and/or short oral corticosteroid bursts are recommended. For moderate-to-severe PANS, oral or intravenous corticosteroids may be sufficient. However, intravenous immunoglobulin (IVIG) is often the preferred treatment for these patients by most PRC members. For more severe or chronic presentations, prolonged corticosteroid courses (with taper) or repeated high-dose corticosteroids may be indicated. For PANS with extreme and life-threatening impairment, therapeutic plasma exchange is the first-line therapy given either alone or in combination with IVIG, high-dose intravenous corticosteroids, and/or rituximab. These recommendations will help guide the use of anti-inflammatory and immunomodulatory therapy in the treatment of PANS.
• Patel, S., Daines, M. O., & Buckley, R. D. (2017). Bacterial meningitis in pediatric patient with C2 deficiency. The Journal of Allergy and Clinical Immunology, 139(2), AB23. doi:10.1016/j.jaci.2016.12.031
• Polley, D. J., Mihara, K., Ramachandran, R., Vliagoftis, H., Renaux, B., Saifeddine, M., Daines, M. O., Boitano, S., & Hollenberg, M. D. (2017). Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase-activated receptor-2. Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, 47(7), 946-960.
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  Allergy to the German cockroach (Blattella germanica) is a significant asthma risk factor for inner-city communities. Cockroach, like other allergens, contains trypsin-like enzyme activity that contributes to allergenicity and airway inflammation by activating proteinase-activated receptors (PARs). To date, the enzymes responsible for the proteolytic activity of German cockroach allergen have not been characterized.
• Polley, D., Mihara, K., Ramachandran, R., Vliagoftis, H., Renaux, B., Saifeddine, M., Daines, M. O., Boitano, S., & Hollenberg, H. (2017). Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase-activated receptor-2 (PAR2). Clinical and Experimental Allergy, Accepted for publication in 2017.
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  BACKGROUND: Allergy to the German cockroach (Blattella germanica) is a significant asthma risk factor for inner-city communities. Cockroach, like other allergens, contains trypsin-like enzyme activity that contributes to allergenicity and airway inflammation by activating proteinase-activated receptors (PARs). To date, the enzymes responsible for the proteolytic activity of German cockroach allergen have not been characterized. OBJECTIVES: We aimed to identify, isolate and characterize the trypsin-like proteinases in a German cockroach allergen extract used for clinical skin tests. For each enzyme, we sought to determine (1) its substrate and inhibitor enzyme kinetics (Km and IC50); (2) its amino acid sequence and (3) its ability to activate calcium signaling and/or ERK1/2 phosphorylation via PAR2. METHODS: Using a trypsin-specific activity-based probe, we detected three distinct enzymes that were isolated using ion-exchange chromatography. Each enzyme was sequenced by mass spectometery (deconvoluted with an expressed sequence tag library), evaluated kinetically for its substrate/inhibitor profile and assessed for its ability to activate PAR2 signaling. FINDINGS: Each of the three serine proteinase-activity-based probe-labelled enzymes isolated were biochemically distinct, with different enzyme kinetic profiles and primary amino acid sequences. The three enzymes showed a 57 to 71% sequence identity with a proteinase previously cloned from the American cockroach (Per a 10). Each enzyme was found to activate both Ca++ and MAPK signaling via PAR2. CONCLUSIONS AND RELEVANCE: We have identified three distinct allergen proteinases from the German cockroach that may play different roles for allergen-sensitization in vivo via PAR2 and may represent attractive therapeutic targets for asthma.
• Rice, S. A., Cabrera, A., Daines, M. O., Spiekerman, C., & Klinger, P. (2020). A Case Study of Acute Onset Obsessive-Compulsive Symptoms in a Pediatric Patient. Journal of Multidisciplinary Clinical Case Reports, 1a(103), 1-6. doi:doi: https://doi.org/10.33790/ jmccr1100103
• Addison, K., Morse, J., Robichaud, A., Daines, M. O., & Ledford, J. (2016). A novel method to test bronchodilators in vivo.. Journal of Infectious Pulmonary Diseases.
• Daines, M. O., Thyne, S., Phipatanakul, W., Wechsler, M. E., Sorkness, C. A., Ross, K. R., Robison, R. G., Raissy, H. H., Peters, S. P., Olin, J. T., Myers, R., Morgan, W. J., Marbin, J., Ly, N. P., Lima, J. J., Lazarus, S. C., Lang, J. E., Kumar, H. V., Israel, E., , Gower, W. A., et al. (2016). Acetaminophen versus Ibuprofen in Young Children with Mild Persistent Asthma. The New England Journal of Medicine. doi:10.1056/nejmoa1515990
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  Studies have suggested an association between frequent acetaminophen use and asthma-related complications among children, leading some physicians to recommend that acetaminophen be avoided in children with asthma; however, appropriately designed trials evaluating this association in children are lacking.In a multicenter, prospective, randomized, double-blind, parallel-group trial, we enrolled 300 children (age range, 12 to 59 months) with mild persistent asthma and assigned them to receive either acetaminophen or ibuprofen when needed for the alleviation of fever or pain over the course of 48 weeks. The primary outcome was the number of asthma exacerbations that led to treatment with systemic glucocorticoids. Children in both groups received standardized asthma-controller therapies that were used in a simultaneous, factorially linked trial.Participants received a median of 5.5 doses (interquartile range, 1.0 to 15.0) of trial medication; there was no significant between-group difference in the median number of doses received (P=0.47). The number of asthma exacerbations did not differ significantly between the two groups, with a mean of 0.81 per participant with acetaminophen and 0.87 per participant with ibuprofen over 46 weeks of follow-up (relative rate of asthma exacerbations in the acetaminophen group vs. the ibuprofen group, 0.94; 95% confidence interval, 0.69 to 1.28; P=0.67). In the acetaminophen group, 49% of participants had at least one asthma exacerbation and 21% had at least two, as compared with 47% and 24%, respectively, in the ibuprofen group. Similarly, no significant differences were detected between acetaminophen and ibuprofen with respect to the percentage of asthma-control days (85.8% and 86.8%, respectively; P=0.50), use of an albuterol rescue inhaler (2.8 and 3.0 inhalations per week, respectively; P=0.69), unscheduled health care utilization for asthma (0.75 and 0.76 episodes per participant, respectively; P=0.94), or adverse events.Among young children with mild persistent asthma, as-needed use of acetaminophen was not shown to be associated with a higher incidence of asthma exacerbations or worse asthma control than was as-needed use of ibuprofen. (Funded by the National Institutes of Health; AVICA ClinicalTrials.gov number, NCT01606319.).
• Daines, M. O., Wright, A. L., Wright, A. L., Stern, D. A., Mathur, A. K., Martinez, F. D., Daines, M. O., & Carr, T. F. (2016). Sensitivity and Specificity of a Clinical Diagnosis of Allergic Rhinitis in Childhood. The Journal of Allergy and Clinical Immunology, 137(2), AB163. doi:10.1016/j.jaci.2015.12.664
• Fitzpatrick, A. M., Jackson, D. J., Mauger, D. T., Boehmer, S. J., Phipatanakul, W., Sheehan, W. J., Moy, J. N., Paul, I. M., Bacharier, L. B., Cabana, M. D., Covar, R., Holguin, F., Lemanske, R. F., Martinez, F. D., Pongracic, J. A., Beigelman, A., Baxi, S. N., Benson, M., Blake, K., , Chmiel, J. F., et al. (2016). Individualized therapy for persistent asthma in young children. The Journal of allergy and clinical immunology, 138(6), 1608-1618.e12.
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  Phenotypic presentations in young children with asthma are varied and might contribute to differential responses to asthma controller medications.
• Sheehan, W. J., Mauger, D. T., Paul, I. M., Moy, J. N., Boehmer, S. J., Szefler, S. J., Fitzpatrick, A. M., Jackson, D. J., Bacharier, L. B., Cabana, M. D., Covar, R., Holguin, F., Lemanske, R. F., Martinez, F. D., Pongracic, J. A., Beigelman, A., Baxi, S. N., Benson, M., Blake, K., , Chmiel, J. F., et al. (2016). Acetaminophen versus Ibuprofen in Young Children with Mild Persistent Asthma. The New England journal of medicine, 375(7), 619-30.
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  Studies have suggested an association between frequent acetaminophen use and asthma-related complications among children, leading some physicians to recommend that acetaminophen be avoided in children with asthma; however, appropriately designed trials evaluating this association in children are lacking.
• Morgan, W., Daines, M., & Cheah, C. (2015). Proinflammatory Environmental Exposures in Asthma. The FASEB Journal, 29(S1). doi:10.1096/fasebj.29.1_supplement.571.8
• Soler, X., Searing, D. A., Santiago, M. T., Morgan, W. J., Knox, K. S., Goodwin, J. L., Ezmigna, D. R., Daines, M. O., Berry, C. E., Zheng, G., Zagaja, V., Yasin, R., Xu, B., Wu, N., Wu, E. Y., Wolf, D., Wise, R. A., Williams, J., Wences, J. A., , Welter, J., et al. (2015). Effect of a soy isoflavone supplement on lung function and clinical outcomes in patients with poorly controlled asthma: a randomized clinical trial.. JAMA, 313(20), 2033-43. doi:10.1001/jama.2015.5024
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  Soy isoflavone supplements are used to treat several chronic diseases, although the data supporting their use are limited. Some data suggest that supplementation with soy isoflavone may be an effective treatment for patients with poor asthma control..To determine whether a soy isoflavone supplement improves asthma control in adolescent and adult patients with poorly controlled disease..Multicenter, randomized, double-blind, placebo-controlled trial conducted between May 2010 and August 2012 at 19 adult and pediatric pulmonary and allergy centers in the American Lung Association Asthma Clinical Research Centers network. Three hundred eighty-six adults and children aged 12 years or older with symptomatic asthma while taking a controller medicine and low dietary soy intake were randomized, and 345 (89%) completed spirometry at week 24..Participants were randomly assigned to receive soy isoflavone supplement containing 100 mg of total isoflavones (n=193) or matching placebo (n=193) in 2 divided doses administered daily for 24 weeks..The primary outcome measure was change in forced expiratory volume in the first second (FEV1) at 24 weeks. Secondary outcome measures were symptoms, episodes of poor asthma control, Asthma Control Test score (range, 5-25; higher scores indicate better control), and systemic and airway biomarkers of inflammation..Mean changes in prebronchodilator FEV1 over 24 weeks were 0.03 L (95% CI, -0.01 to 0.08 L) in the placebo group and 0.01 L (95% CI, -0.07 to 0.07 L) in the soy isoflavone group, which were not significantly different (P = .36). Mean changes in symptom scores on the Asthma Control Test (placebo, 1.98 [95% CI, 1.42-2.54] vs soy isoflavones, 2.20 [95% CI, 1.53-2.87]; positive values indicate a reduction in symptoms), number of episodes of poor asthma control (placebo, 3.3 [95% CI, 2.7-4.1] vs soy isoflavones, 3.0 [95% CI, 2.4-3.7]), and changes in exhaled nitric oxide (placebo, -3.48 ppb [95% CI, -5.99 to -0.97 ppb] vs soy isoflavones, 1.39 ppb [95% CI, -1.73 to 4.51 ppb]) did not significantly improve more with the soy isoflavone supplement than with placebo. Mean plasma genistein level increased from 4.87 ng/mL to 37.67 ng/mL (P < .001) in participants receiving the supplement..Among adults and children aged 12 years or older with poorly controlled asthma while taking a controller medication, use of a soy isoflavone supplement, compared with placebo, did not result in improved lung function or clinical outcomes. These findings suggest that this supplement should not be used for patients with poorly controlled asthma..clinicaltrials.gov Identifier: NCT01052116.
• Sherwood, C. L., Daines, M. O., Price, T. J., Vagner, J., & Boitano, S. (2014). A highly potent agonist to protease-activated receptor-2 reveals apical activation of the airway epithelium resulting in Ca2+-regulated ion conductance. American journal of physiology. Cell physiology, 307(8), C718-26.
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  The airway epithelium provides a barrier that separates inhaled air and its various particulates from the underlying tissues. It provides key physiological functions in both sensing the environment and initiating appropriate innate immune defenses to protect the lung. Protease-activated receptor-2 (PAR2) is expressed both apically and basolaterally throughout the airway epithelium. One consequence of basolateral PAR2 activation is the rapid, Ca(2+)-dependent ion flux that favors secretion in the normally absorptive airway epithelium. However, roles for apically expressed PAR2 activation have not been demonstrated, in part due to the lack of specific, high-potency PAR2 ligands. In the present study, we used the newly developed PAR2 ligand 2at-LIGRLO(PEG3-Pam)-NH2 in combination with well-differentiated, primary cultured airway epithelial cells from wild-type and PAR2 (-/-) mice to examine the physiological role of PAR2 in the conducting airway after apical activation. Using digital imaging microscopy of intracellular Ca(2+) concentration changes, we verified ligand potency on PAR2 in primary cultured airway cells. Examination of airway epithelial tissue in an Ussing chamber showed that apical activation of PAR2 by 2at-LIGRLO(PEG3-Pam)-NH2 resulted in a transient decrease in transepithelial resistance that was due to increased apical ion efflux. We determined pharmacologically that this increase in ion conductance was through Ca(2+)-activated Cl(-) and large-conductance K(+) channels that were blocked with a Ca(2+)-activated Cl(-) channel inhibitor and clotrimazole, respectively. Stimulation of Cl(-) efflux via PAR2 activation at the airway epithelial surface can increase airway surface liquid that would aid in clearing the airway of noxious inhaled agents.
• Daines, M. O., Hollenberg, M. D., Boitano, S., Vliagoftis, H., Ramachandran, R., Mihara, K., & Polley, D. B. (2013). Cockroach Allergen Proteinases Regulate Proteinase‐Activated Receptor‐1 (PAR1) Signalling. The FASEB Journal. doi:10.1096/fasebj.27.1_supplement.1171.6
• Daines, M. O., Warrier, M. R., Chen, W., Tabata, Y., Khurana, G. K., & Gibson, M. (2013). Soluble Forms 2 Yields Distinct Membrane and α Receptor Alternative Splicing of IL-13. JACI.
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  HersheyM. Gibson, Michael O. Daines and Gurjit K. Khurana Yasuhiro Tabata, Weiguo Chen, Manoj R. Warrier, Aaronhttp://www.jimmunol.org/content/177/11/7905J Immunol€2006; 177:7905-7912; ;Referenceshttp://www.jimmunol.org/content/177/11/7905.full#ref-list-1This article cites 29 articles, 16 of which you can access for free at: Subscriptionshttp://jimmunol.org/subscriptionsInformation about subscribing to The Journal of Immunology is online at: Permissionshttp://www.aai.org/ji/copyright.htmlSubmit copyright permission requests at: Email Alertshttp://jimmunol.org/cgi/alerts/etocReceive free email-alerts when new articles cite this article. Sign up at:
• Daines, M., Polley, D., Mihara, K., Ramachandran, R., Vliagoftis, H., Boitano, S., & Hollenberg, M. D. (2013). Cockroach Allergen Proteinases Regulate Proteinase‐Activated Receptor‐1 (PAR1) Signalling. The FASEB Journal, 27(S1). doi:10.1096/fasebj.27.1_supplement.1171.6
• Graham, L. V., Flynn, A. N., Sherwood, C. L., Schultz, S. M., Hoffman, J., Daines, M. O., Daines, M. O., Boitano, S. A., & Graham, L. V. (2013). Measuring A. alternata Protease Activity and Their Effects on Asthma. Honors Thesis.
• Daines, M. O., Hollenberg, M. D., Boitano, S., Vliagoftis, H., Renaux, B., Saifeddine, M., Mihara, K., & Polley, D. B. (2012). Allergen‐derived proteinases: Isolation, characterization and signaling via proteinase‐activated receptors (PARs). The FASEB Journal. doi:10.1096/fasebj.26.1_supplement.664.10
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  Objective To identify proteolytic enzymes in insect and mould asthma allergens and assess their ability to signal via the PARs. Methods Allergen-derived trypsin-like enzymes known to contribute to lung inflammation were visualized (western blot) with serine proteinase activity-based probes (ABP). Enzymes were purified using ion-exchange chromatography and analyzed for their ABP reactivity & substrate-inhibitor kinetic profiles. Each enzyme was tested for its activation of PAR–dependent calcium signaling in PAR1 & PAR2-expressing KNRK cells. ABP-labeled enzymes were isolated by avidin affinity chromatography and their peptide sequences determined (mass spectrometry). Results Both allergens contained ABP-labeled proteinases with MWs of 20–26 kDa. Each enzyme had distinct kinetic substrate (Km) and inhibitor (Ki) profiles. The enzymes all caused PAR2 calcium signaling, and differentially activated PAR1. Sequencing revealed that the allergen proteinases were homologous with previously described trypsin-like enzymes. Conclusions Diverse allergens contain multiple biochemically distinct trypsin-like proteinases that are able to target PARs 1 and 2. Thus, PARs and their activating allergen proteinases may be attractive therapeutic targets for the treatment of allergen-induced asthma. Support: Canadian Institutes of Health Research, NIH USA and Lung Association of Alberta & NWT
• Boitano, S., Flynn, A. N., Sherwood, C. L., Schulz, S. M., Hoffman, J., Gruzinova, I., & Daines, M. O. (2011). Alternaria alternata serine proteases induce lung inflammation and airway epithelial cell activation via PAR2. American journal of physiology. Lung cellular and molecular physiology, 300(4), L605-14.
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  Allergens are diverse proteins from mammals, birds, arthropods, plants, and fungi. Allergens associated with asthma (asthmagens) share a common protease activity that may directly impact respiratory epithelial biology and lead to symptoms of asthma. Alternaria alternata is a strong asthmagen in semiarid regions. We examined the impact of proteases from A. alternata on lung inflammation in vivo and on cleaving protease-activated receptor-2 (PAR(2)) in vitro. A. alternata filtrate applied to the airway in nonsensitized Balb/c mice induced a protease-dependent lung inflammation. Moreover, A. alternata filtrate applied to human bronchial epithelial cells (16HBE14o-) induced changes in intracellular Ca(2+) concentration ([Ca(2+)](i)), consistent with PAR(2) activation. These effects were blocked by heat inactivation or by serine protease inhibition of A. alternata filtrates, and mimicked by PAR(2) specific ligands SLIGRL-NH(2) or 2-furoyl-LIGRLO-NH(2), but not the PAR(1)-specific ligand TFLLR-NH(2). Desensitization of PAR(2) in 16HBE14o- cells with 2-furoyl-LIGRLO-NH(2) or trypsin prevented A. alternata-induced [Ca(2+)](i) changes while desensitization of PAR(1), PAR(3), and PAR(4) with thrombin had no effect on A. alternata-induced Ca(2+) responses. Furthermore, the Ca(2+) response to A. alternata filtrates was dependent on PAR(2) expression in stably transfected HeLa cell models. These data demonstrate that A. alternata proteases act through PAR(2) to induce rapid increases in human airway epithelial [Ca(2+)](i) in vitro and cell recruitment in vivo. These responses are likely critical early steps in the development of allergic asthma.
• Daines, M. O., Gruzinova, I., Hoffman, J., Schulz, S. M., Sherwood, C. L., Flynn, A. N., & Boitano, S. (2011). Alternaria alternataserine proteases induce lung inflammation and airway epithelial cell activation via PAR₂. American Journal of Physiology-lung Cellular and Molecular Physiology, 300(4), L605-L614. doi:10.1152/ajplung.00359.2010
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  Allergens are diverse proteins from mammals, birds, arthropods, plants, and fungi. Allergens associated with asthma (asthmagens) share a common protease activity that may directly impact respiratory epithelial biology and lead to symptoms of asthma. Alternaria alternata is a strong asthmagen in semiarid regions. We examined the impact of proteases from A. alternata on lung inflammation in vivo and on cleaving protease-activated receptor-2 (PAR(2)) in vitro. A. alternata filtrate applied to the airway in nonsensitized Balb/c mice induced a protease-dependent lung inflammation. Moreover, A. alternata filtrate applied to human bronchial epithelial cells (16HBE14o-) induced changes in intracellular Ca(2+) concentration ([Ca(2+)](i)), consistent with PAR(2) activation. These effects were blocked by heat inactivation or by serine protease inhibition of A. alternata filtrates, and mimicked by PAR(2) specific ligands SLIGRL-NH(2) or 2-furoyl-LIGRLO-NH(2), but not the PAR(1)-specific ligand TFLLR-NH(2). Desensitization of PAR(2) in 16HBE14o- cells with 2-furoyl-LIGRLO-NH(2) or trypsin prevented A. alternata-induced [Ca(2+)](i) changes while desensitization of PAR(1), PAR(3), and PAR(4) with thrombin had no effect on A. alternata-induced Ca(2+) responses. Furthermore, the Ca(2+) response to A. alternata filtrates was dependent on PAR(2) expression in stably transfected HeLa cell models. These data demonstrate that A. alternata proteases act through PAR(2) to induce rapid increases in human airway epithelial [Ca(2+)](i) in vitro and cell recruitment in vivo. These responses are likely critical early steps in the development of allergic asthma.
• Daines, M. O., Hershey, G. K., Silverman, G. A., Wills-Karp, M., Cras, T. D., Whitsett, J. A., Wert, S. E., Stringer, K. F., Chakir, J., Bass, S. A., Brandt, E. B., Stier, M. T., Gibson, A. M., Ericksen, M. B., Askew, D. J., & Sivaprasad, U. (2011). A nonredundant role for mouse Serpinb3a in the induction of mucus production in asthma. The Journal of Allergy and Clinical Immunology. doi:10.1016/j.jaci.2010.10.009
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  Asthma is a major public health burden worldwide. Studies from our group and others have demonstrated that SERPINB3 and SERPINB4 are induced in patients with asthma; however, their mechanistic role in asthma has yet to be determined.To evaluate the role of Serpin3a, the murine homolog of SERPINB3 and SERPINB4, in asthma.We studied wild-type Balb/c and Serpinb3a-null mice in house dust mite or IL-13-induced asthma models and evaluated airway hyperresponsiveness, inflammation, and goblet cell hyperplasia.Airway hyperresponsiveness and goblet cell hyperplasia were markedly attenuated in the Serpinb3a-null mice compared with the wild-type mice after allergen challenge, with minimal effects on inflammation. Expression of sterile alpha motif pointed domain containing v-ets avian erythroblastosis virus E26 oncogene homolog transcription factor (SPDEF), a transcription factor that mediates goblet cell hyperplasia, was decreased in the absence of Serpinb3a. IL-13-treated Serpinb3a-null mice showed attenuated airway hyperresponsiveness, inflammation, and mucus production.Excessive mucus production and mucus plugging are key pathologic features of asthma, yet the mechanisms responsible for mucus production are not well understood. Our data reveal a novel nonredundant role for Serpinb3a in mediating mucus production through regulation of SPDEF expression. This pathway may be used to target mucus hypersecretion effectively.
• Daines, M. O., Shultz, S., Sherwood, C. L., Hoffman, J., Gruzinova, I., Flynn, A. N., Daines, M. O., & Boitano, S. (2011). PAR-2 Activation by Alternaria alternata Proteases Induces Airway Epithelial Cell Activation and Lung Inflammation. The Journal of Allergy and Clinical Immunology, 127(2), AB59-AB59. doi:10.1016/j.jaci.2010.12.246
• Muralidhar, A., Borbon, I. A., Esharif, D. M., Ke, W., Manacheril, R., Daines, M., & Erickson, R. P. (2011). Pulmonary function and pathology in hydroxypropyl-beta-cyclodextin-treated and untreated Npc1⁻/⁻ mice. Molecular genetics and metabolism, 103(2), 142-7.
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  Lung dysfunction is an important part of the pathology of the neurodegenerative disorder, Niemann-Pick C1 (NPC1). We have studied the pulmonary disease in the Npc1(NIH/NIH) mouse model. On histology, we find large numbers of alveolar foamy macrophages but no alveolar proteinosis. Lung weight as percent of body weight was markedly increased; using the flexiVent small animal ventilator (SCIREQ, Inc.), we find inspiratory capacity, elastance and hysterisivity to be increased while resistance was not changed. Cholesterol measurements show a doubling of lung cholesterol levels. Collagen is also increased. Treatment of Npc1(-/-) mice with hydroxypropyl-β-cyclodextrin (HPBCD), despite efficacious effects in brain and liver, results in little difference from age-matched controls (using a CNS-expressed transgene to extend the life expectancy of the Npc1(-/-) mice) for these variables.
• Sivaprasad, U., Askew, D. J., Ericksen, M. B., Gibson, A. M., Stier, M. T., Brandt, E. B., Bass, S. A., Daines, M. O., Chakir, J., Stringer, K. F., Wert, S. E., Whitsett, J. A., Le Cras, T. D., Wills-Karp, M., Silverman, G. A., & Khurana Hershey, G. K. (2011). A nonredundant role for mouse Serpinb3a in the induction of mucus production in asthma. The Journal of allergy and clinical immunology, 127(1), 254-61, 261.e1-6.
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  Asthma is a major public health burden worldwide. Studies from our group and others have demonstrated that SERPINB3 and SERPINB4 are induced in patients with asthma; however, their mechanistic role in asthma has yet to be determined.
• Winzerling, J. J., Daines, M., Winzerling, J. J., Thomson, C. A., Thompson, P. A., Stendell-hollis, N. R., & Daines, M. O. (2011). Adherence to a Mediterranean-style diet and change in inflammatory biomarkers and fatty acid profiles in lactating women. The FASEB Journal, 25.
• Pivniouk, V. I., Daines, M. O., Vercelli, D., Wlasiuk, G., Vercelli, D., Strempel, J. M., Rosenbaum, D., Pivniouk, V., Pivniouk, O., Kim, K., Kiesler, P., Daines, M. O., & Bailey, T. J. (2010). Faithful epigenetic and transcriptional regulation of a transgenic human Th2 cytokine gene locus in the murine nuclear environment. Journal of Immunology, 184.
• Wills-karp, M., Whitsett, J. A., Wert, S. E., Stringer, K. F., Stier, M. T., Sivaprasad, U., Silverman, G. A., Lindsey, M., Lecras, T. D., Hershey, G. K., Gibson, A. M., Ericksen, M. B., Daines, M. O., Chakir, J., Brandt, E. B., Bass, S. A., Askew, D. S., & Aronow, B. J. (2010). A non-redundant role for Serpinb3a in the induction of mucus production in asthma. Journal of Immunology, 184.
• Winzerling, J. J., Daines, M., Winzerling, J. J., Thomson, C. A., Thompson, P. A., Stendell-hollis, N. R., Laudermilk, M., & Daines, M. O. (2010). A Mediterranean diet intervention rich in walnuts among lactating women: study design and baseline characteristics. The FASEB Journal, 24.
• Chen, W., Tabata, Y., Gibson, A. M., Daines, M. O., Warrier, M. R., Wills-Karp, M., & Hershey, G. K. (2008). Matrix metalloproteinase 8 contributes to solubilization of IL-13 receptor alpha2 in vivo. The Journal of allergy and clinical immunology, 122(3), 625-32.
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  IL-13 receptor alpha2 (IL-13R alpha 2) is a high-affinity receptor for IL-13, a central mediator of allergic asthma. It acts predominantly as a decoy receptor but can also contribute to IL-13 responses under certain conditions. IL-13R alpha 2 exists in soluble and membrane forms, which can both bind IL-13 and modulate its activity. Yet the proteolytic processes that contribute to the generation of soluble IL-13R alpha 2 are largely unknown.
• Halonen, M., Daines, M. O., Halonen, M., Daines, M. O., Campbell, R., & Boitano, S. (2008). Effect of Proteases and Anti-proteases on Alternaria-Induced Inflammation. The FASEB Journal, 22(S2), 650-650. doi:10.1096/fasebj.22.2_supplement.650
• Marsh, R. A., Lucky, A. W., Walsh, T. J., Pacheco, M. C., Rinaldi, M. G., Mailler-Savage, E., Puel, A., Casanova, J. L., Bleesing, J. J., Filippi, M. D., Williams, D. A., Daines, M. O., & Filipovich, A. H. (2008). Cutaneous infection with Metarhizium anisopliae in a patient with hypohidrotic ectodermal dysplasia and immune deficiency. The Pediatric infectious disease journal, 27(3), 283-4.
• Daines, M. O., Chen, W., Tabata, Y., Walker, B. A., Gibson, A. M., Masino, J. A., Warrier, M. R., Daines, C. L., Wenzel, S. E., & Hershey, G. K. (2007). Allergen-dependent solubilization of IL-13 receptor alpha2 reveals a novel mechanism to regulate allergy. The Journal of allergy and clinical immunology, 119(2), 375-83.
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  Allergic sensitization affects half of western populations and often precedes the development of allergic disorders including asthma. Despite the critical role of allergens in the pathogenesis of these disorders, little is known about how allergens modulate the immune response. IL-13 receptor alpha2 (IL-13Ralpha2) is a decoy receptor for IL-13.
• Zornes, P. A., Hershey, G. K., & Daines, M. O. (2007). Signaling Interactions Between IL-4 or IL-13 and Leukotrienes in Allergic Inflammation. The Journal of Allergy and Clinical Immunology, 119(1), S288. doi:10.1016/j.jaci.2006.12.500
• Daines, M. O., Hershey, G. K., Basu, S., Warrier, M., Chen, W., Walker, B. A., & Tabata, Y. (2006). Level of Expression of IL-13Rα2 Impacts Receptor Distribution and IL-13 Signaling. Journal of Immunology. doi:10.4049/jimmunol.176.12.7495
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  IL-13, a critical cytokine for allergic inflammation, exerts its effects through a complex receptor system including IL-4Ralpha, IL-13Ralpha1, and IL-13Ralpha2. IL-4Ralpha and IL-13Ralpha1 form a heterodimeric signaling receptor for IL-13. In contrast, IL-13Ralpha2 binds IL-13 with high affinity but does not signal. IL-13Ralpha2 exists on the cell surface, intracellularly, and in soluble form, but no information is available regarding the relative distributions of IL-13Ralpha2 among these compartments, whether the compartments communicate, and how the relative expression levels impact IL-13 responses. Herein, we investigated the distribution of IL-13Ralpha2 in transfected and primary cells, and we evaluated how the total level of IL-13Ralpha2 expression impacted its distribution. Our results demonstrate that the distribution of IL-13Ralpha2 is independent of the overall level of expression. The majority of the IL-13Ralpha2 protein existed in intracellular pools. Surface IL-13Ralpha2 was continually released into the medium in a soluble form, yet surface expression remained constant supporting receptor trafficking to the cell surface. IL-13Ralpha2 inhibited IL-13 signaling proportionally to its level of expression, and this inhibition could be overcome with high concentrations of IL-13.
• Daines, M. O., Hershey, G. K., Gibson, A. M., Warrier, M., Chen, W., & Tabata, Y. (2006). Allergy-Driven Alternative Splicing of IL-13 Receptor α2 Yields Distinct Membrane and Soluble Forms. Journal of Immunology. doi:10.4049/jimmunol.177.11.7905
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  IL-13 is a key mediator of allergic inflammation. Its diverse functions are mediated by a complex receptor system including IL-4Ralpha, IL-13Ralpha1, and IL-13Ralpha2. IL-4Ralpha and IL-13Ralpha1 form a high-affinity signaling heterodimer. IL-13Ralpha2 binds IL-13 with high affinity and has been found to exist in membrane and soluble forms. Soluble IL-13Ralpha2 has been postulated as a critical endogenous modulator of IL-13 responses. However, the mechanism of generation for the soluble form remains unclear. We present the initial study that a mechanism for generation of the soluble form is alternative splicing and that alternative splicing yields a distinct form of soluble IL-13Ralpha2. We found that several mouse organs expressed two IL-13Ralpha2 transcripts, the 1152-bp transcript encoding the full-length protein and the 1020-bp transcript lacking exon10, which encodes the transmembrane region. Deletion of exon 10 (DeltaEx10) caused a frameshift resulting in a different amino acid sequence from position 327 to position 339 and early termination. Constructs encoding both splice variants were transfected into WEHI-274.1 cells. Transfectants expressing the full-length transcript had IL-13Ralpha2 on the cell surface but produced minimal soluble IL-13Ralpha2 in the supernatants. In contrast, transfectants expressing the DeltaEx10 transcript displayed no membrane IL-13Ralpha2 but secreted high levels of soluble IL-13Ralpha2 capable of inhibiting IL-13 signaling. Both variants bound IL-13, but the DeltaEx10 variant displayed approximately 2-fold increase in IL-13 binding activity. Expression of the two IL-13Ralpha2 transcripts was differentially regulated in vivo in an experimental allergic asthma model. Thus, alternatively spliced variants of IL-13Ralpha2 may have a distinct biologic function in vivo.
• Daines, M. O., Tabata, Y., Walker, B. A., Chen, W., Warrier, M. R., Basu, S., & Hershey, G. K. (2006). Level of expression of IL-13R alpha 2 impacts receptor distribution and IL-13 signaling. Journal of immunology (Baltimore, Md. : 1950), 176(12), 7495-501.
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  IL-13, a critical cytokine for allergic inflammation, exerts its effects through a complex receptor system including IL-4Ralpha, IL-13Ralpha1, and IL-13Ralpha2. IL-4Ralpha and IL-13Ralpha1 form a heterodimeric signaling receptor for IL-13. In contrast, IL-13Ralpha2 binds IL-13 with high affinity but does not signal. IL-13Ralpha2 exists on the cell surface, intracellularly, and in soluble form, but no information is available regarding the relative distributions of IL-13Ralpha2 among these compartments, whether the compartments communicate, and how the relative expression levels impact IL-13 responses. Herein, we investigated the distribution of IL-13Ralpha2 in transfected and primary cells, and we evaluated how the total level of IL-13Ralpha2 expression impacted its distribution. Our results demonstrate that the distribution of IL-13Ralpha2 is independent of the overall level of expression. The majority of the IL-13Ralpha2 protein existed in intracellular pools. Surface IL-13Ralpha2 was continually released into the medium in a soluble form, yet surface expression remained constant supporting receptor trafficking to the cell surface. IL-13Ralpha2 inhibited IL-13 signaling proportionally to its level of expression, and this inhibition could be overcome with high concentrations of IL-13.
• Hershey, G. K., Ericksen, M. B., Ericksen, E., & Daines, M. O. (2006). Stat6 Nuclear Export Elements are Unique from other Stat Family Members. The Journal of Allergy and Clinical Immunology, 117(2), S143. doi:10.1016/j.jaci.2005.12.571
• Tabata, Y., Chen, W., Warrier, M. R., Gibson, A. M., Daines, M. O., & Hershey, G. K. (2006). Allergy-driven alternative splicing of IL-13 receptor alpha2 yields distinct membrane and soluble forms. Journal of immunology (Baltimore, Md. : 1950), 177(11), 7905-12.
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  IL-13 is a key mediator of allergic inflammation. Its diverse functions are mediated by a complex receptor system including IL-4Ralpha, IL-13Ralpha1, and IL-13Ralpha2. IL-4Ralpha and IL-13Ralpha1 form a high-affinity signaling heterodimer. IL-13Ralpha2 binds IL-13 with high affinity and has been found to exist in membrane and soluble forms. Soluble IL-13Ralpha2 has been postulated as a critical endogenous modulator of IL-13 responses. However, the mechanism of generation for the soluble form remains unclear. We present the initial study that a mechanism for generation of the soluble form is alternative splicing and that alternative splicing yields a distinct form of soluble IL-13Ralpha2. We found that several mouse organs expressed two IL-13Ralpha2 transcripts, the 1152-bp transcript encoding the full-length protein and the 1020-bp transcript lacking exon10, which encodes the transmembrane region. Deletion of exon 10 (DeltaEx10) caused a frameshift resulting in a different amino acid sequence from position 327 to position 339 and early termination. Constructs encoding both splice variants were transfected into WEHI-274.1 cells. Transfectants expressing the full-length transcript had IL-13Ralpha2 on the cell surface but produced minimal soluble IL-13Ralpha2 in the supernatants. In contrast, transfectants expressing the DeltaEx10 transcript displayed no membrane IL-13Ralpha2 but secreted high levels of soluble IL-13Ralpha2 capable of inhibiting IL-13 signaling. Both variants bound IL-13, but the DeltaEx10 variant displayed approximately 2-fold increase in IL-13 binding activity. Expression of the two IL-13Ralpha2 transcripts was differentially regulated in vivo in an experimental allergic asthma model. Thus, alternatively spliced variants of IL-13Ralpha2 may have a distinct biologic function in vivo.
• Warrier, M. R., Hershey, G. K., Gibson, A. M., Daines, M. O., Cunningham, C. M., & Chen, W. (2006). IL-13 Receptor Alpha 2 Lung-transgenic Mice Displayed Reduced Airway Hyperresponsiveness and Inflammation in a House Dust Mite Challenged Model. The Journal of Allergy and Clinical Immunology, 117(2), S150. doi:10.1016/j.jaci.2005.12.599
• Warrier, M. R., Tabata, Y., Hershey, G. K., Daines, M. O., & Chen, W. (2006). Expression of IL-13 Receptors in Human Airway and Skin Cells. The Journal of Allergy and Clinical Immunology, 117(2), S255. doi:10.1016/j.jaci.2005.12.1008
• Warrier, M. R., Tabata, Y., Hershey, G. K., Daines, M. O., & Chen, W. (2006). IL-13 Receptors in the Skin. The Journal of Allergy and Clinical Immunology, 117(2), S248. doi:10.1016/j.jaci.2005.12.983
• Warrier, M. R., Walker, W., Tabata, Y., Masino, J. A., Hershey, G. K., Gibson, A. M., Daines, M. O., & Chen, W. (2006). Distribution of IL-13Ra2: Impact of Allergen Exposure and Level of Expression. The Journal of Allergy and Clinical Immunology, 117(2), S148. doi:10.1016/j.jaci.2005.12.589
• Boesch, R. P., Daines, M., Kaul, A., Cotton, R., & Amin, R. (2005). Lymphoproliferative disorder of the airway of an adolescent without immunodeficiency. International journal of pediatric otorhinolaryngology, 69(11), 1591-4.
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  Epstein-Barr virus (EBV) is a ubiquitous viral pathogen in humans that has a unique ability to immortalize B-cells. In immunosuppressed individuals, EBV can produce non-neoplastic lymphoproliferative disorders involving various organs. We describe a case report of EBV-associated lymphoproliferative disorder in an immunocompetent 14-year-old male. The case provides a description of EBV-associated lymphoproliferation affecting the upper and lower respiratory tract. The massive submucosal infiltration of B-cells in the lingual tonsils, trachea, and bronchi produced near-complete airway obstruction resulting in tracheotomy. Neither surgical reduction of lingual tonsils nor treatment with steroids was of benefit. An extensive evaluation for immunodeficiency and neoplasia was normal. Treatment with rituximab, an anti-CD20 antibody, resulted in near-complete resolution of the infiltrative process, sufficient to allow decannulation. Rituximab is a treatment option for the rare occurrence of non-neoplastic, EBV-associated, lymphoproliferative disorders.
• Daines, M. O., Amin, R. S., Cotton, R. T., Kaul, A., & Boesch, R. P. (2005). Lymphoproliferative disorder of the airway of an adolescent without immunodeficiency. International Journal of Pediatric Otorhinolaryngology. doi:10.1016/j.ijporl.2005.04.028
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  Epstein-Barr virus (EBV) is a ubiquitous viral pathogen in humans that has a unique ability to immortalize B-cells. In immunosuppressed individuals, EBV can produce non-neoplastic lymphoproliferative disorders involving various organs. We describe a case report of EBV-associated lymphoproliferative disorder in an immunocompetent 14-year-old male. The case provides a description of EBV-associated lymphoproliferation affecting the upper and lower respiratory tract. The massive submucosal infiltration of B-cells in the lingual tonsils, trachea, and bronchi produced near-complete airway obstruction resulting in tracheotomy. Neither surgical reduction of lingual tonsils nor treatment with steroids was of benefit. An extensive evaluation for immunodeficiency and neoplasia was normal. Treatment with rituximab, an anti-CD20 antibody, resulted in near-complete resolution of the infiltrative process, sufficient to allow decannulation. Rituximab is a treatment option for the rare occurrence of non-neoplastic, EBV-associated, lymphoproliferative disorders.
• Guajardo, J. R., Schleifer, K. W., Daines, M. O., Ruddy, R. M., Aronow, B. J., Wills-Karp, M., & Hershey, G. K. (2005). Altered gene expression profiles in nasal respiratory epithelium reflect stable versus acute childhood asthma. The Journal of allergy and clinical immunology, 115(2), 243-51.
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  Asthma is the most common chronic disease of childhood and has a strong genetic component.
• Hershey, G. K., & Daines, M. O. (2005). Structural elements of IL-13Ra2 and the inhibition of IL-13 signaling. The Journal of Allergy and Clinical Immunology, 115(2), S122. doi:10.1016/j.jaci.2004.12.502
• Masino, J. A., Hershey, G. K., & Daines, M. O. (2005). Solubilization of human IL-13Rα 2 following allergen treatment in vitro. The Journal of Allergy and Clinical Immunology, 115(2), S118-S119. doi:10.1016/j.jaci.2004.12.486
• Chen, W., Daines, M. O., & Hershey, G. K. (2004). Methylation of STAT6 modulates STAT6 phosphorylation, nuclear translocation, and DNA-binding activity. Journal of immunology (Baltimore, Md. : 1950), 172(11), 6744-50.
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  Signal transducer and activator of transcription 6 is a transcription factor important for the development of Th2 cells and regulation of gene expression by IL-4 and IL-13. It has been reported that STAT1 activity is regulated by methylation of a conserved arginine residue in the N-terminal domain. Methylation of STAT6 has not yet been explored. We observed methylation of STAT6 in cells transfected with wild-type STAT6, but not in cells transfected with Arg(27)Ala mutant, confirming that STAT6 is methylated on Arg(27). Transfectants expressing mutant Arg(27)Ala STAT6 displayed markedly diminished IL-4-dependent STAT6 phosphorylation and nuclear translocation, and no STAT6 DNA-binding activity compared with wild-type STAT6 transfectants. To confirm this, the experiments were repeated using inhibitors of methylation. In the presence of methylation inhibitors, STAT6 methylation was diminished, as was phosphorylation of STAT6 and STAT6 DNA-binding activity. Thus, methylation is a critical regulator of STAT6 activity, necessary for optimal STAT6 phosphorylation, nuclear translocation, and DNA-binding activity. Furthermore, methylation of STAT6 has distinct effects from those reported with STAT1.
• Chen, W., Daines, M. O., & Khurana Hershey, G. K. (2004). Turning off signal transducer and activator of transcription (STAT): the negative regulation of STAT signaling. The Journal of allergy and clinical immunology, 114(3), 476-89; quiz 490.
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  Signal transducer and activator of transcription (STAT) proteins are a group of transcription factors that transmit signals from the extracellular milieu of cells to the nucleus. They are crucial for the signaling of many cytokines that are mediators of allergic inflammation. Considerable information is known about the activation of STATs and their role in gene transcription; comparably much less is known about how STAT signaling is regulated. Because STATs are critical for the induction of many genes crucial for the allergic cascade and immune host defense, understanding the regulation of these molecules will provide novel insights into allergic and immunodeficiency disorders and will likely identify new targets for therapeutic interventions. This review will summarize the current understanding of the regulation of STAT signaling, emphasizing recent observations.
• Hershey, G. K., Daines, M. O., & Basu, S. (2004). IL-13 signaling in cells stably expressing FLAG-human IL-13R Alpha 2. The Journal of Allergy and Clinical Immunology, 113(2), S324. doi:10.1016/j.jaci.2004.01.672
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  Abstract Rationale IL-13Rα2 is a high affinity receptor for IL-13 that is a possible decoy receptor for IL-13. We developed a model to study the effects of this receptor on IL-13 dependent gene induction. Methods U937 cells are derived from human monocytes and respond to IL-13 by Stat6 signaling and CD23 gene induction. We stably transfected these cells to express a FLAG-IL-13Rα2 fusion protein. Surface expression of FLAG, IL-13Rα2, and IL-13 binding activity were determined in transfected cells and U937 controls. IL-13 dependent Stat6 responses were determined. Results U937 cells transfected to express FLAG-IL-13Rα2 fusion protein were assayed for the presence of FLAG, IL-13Rα2, and IL-13 binding. Transfected cells displayed surface expression of FLAG and IL-13Rα2 by flow cytometry. IL-13 binding was demonstrated in the transfected cells. Untransfected U937 cells demonstrated no significant surface expression of FLAG or IL-13Rα2 and no significant IL-13 binding. Cells expressing FLAG-IL-13Rα2 had decreased activation of Stat6 in response to IL-13 than U937 controls. Conclusions U937 cells transfected with FLAG-IL-13Rα2 fusion protein demonstrate surface expression of this receptor and bind IL-13 as shown by sandwich FACS. Expression of FLAG-IL-13Rα2 decreases IL-13 dependent Stat6 activation.
• Stevenson, M. D., Ruddy, R. M., Hershey, G. K., & Daines, M. O. (2004). Phosphorylated Stat-6 levels in human nasal epithelial (HNE) cells in acute and controlled asthmatic children. The Journal of Allergy and Clinical Immunology, 113(2), S184. doi:10.1016/j.jaci.2004.01.100
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  Abstract Rationale IL-4 and IL-13 signaling via IL-4Rα is a critical pathway in Th-2 mediated disease states such as asthma. The subsequent phosphorylation of Stat-6 is specific for transcription and gene expression induced by these cytokines. HNE cells were selected as a proxy for respiratory epithelium and activated Stat-6 levels were measured and evaluated as a biomarker of asthma. Methods HNE samples were collected via cytobrush sampling of the inferior turbinate from three groups: controlled asthmatic children, children with an asthma exacerbation through the emergency department, and normal healthy children who were skin test negative to common aeroallergens. Cells were incubated with and without IL-4 (20 ng/ml), fixed and permeabilized. A monoclonal antibody specific for phosphorylated Stat-6 site was used to detect the presence of activated Stat-6 by flow cytometry. The percentage change with IL-4 stimulation, corrected for isotype control, was compared between groups via t-test. Results No significant difference between all asthma (n=45) and normals (n=9) was found (p=.97). The difference between controlled asthma (n=23) +1.4% and acute asthma (n=22) +6.1% was not statistically significant (p=0.24) but trended towards a larger percentage change in acute asthma. Conclusions Although not statistically significant, slightly higher levels of phosphorylated Stat-6 were observed in acute asthma versus controlled asthma. This method of measuring a critical signaling molecule in a disease state warrants additional study. Furthermore, this data suggests that activated Stat-6 may have potential as a biomarker in the acute phase of asthma.
• Daines, M. O., Andrews, R. P., Chen, W., El-Zayaty, S. A., & Hershey, G. K. (2003). DNA binding activity of cytoplasmic phosphorylated Stat6 is masked by an interaction with a detergent-sensitive factor. The Journal of biological chemistry, 278(33), 30971-4.
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  Signal transducer and activator of transcription (Stat) 6 is vital to interleukin (IL)-4 and IL-13 responses and the generation of Th2 immunity. We investigated the cellular location of phosphorylated Stat6 and Stat6 DNA binding activity in A201.1 murine B cells and primary splenocytes. Phosphorylated Stat6 was present in cytoplasmic and nuclear extracts from IL-4-treated cells. Confocal microscopy confirmed the presence of phosphorylated Stat6 in the cytoplasm of IL-4-treated cells. In contrast, Stat6 DNA binding activity was present in nuclear extracts, but not in cytoplasmic extracts. Thus, cytoplasmic extracts from IL-4-stimulated cells were devoid of Stat6 DNA binding activity despite the presence of phosphorylated Stat6. Addition of cytoplasmic extracts to nuclear extracts did not inhibit Stat6 DNA binding present in the nuclear extracts. Detergent treatment restored Stat6 DNA binding activity in cytoplasmic extracts of IL-4-stimulated cells. Thus, DNA binding activity of cytoplasmic phosphorylated Stat6 is masked by a factor dissociable by detergent treatment.
• Daines, M. O., Hershey, G. K., & Chen, W. (2003). Arginine 27, a conserved methylation site in Stat6, is essential for IL-4-induced tyrosine-phosphorylation, DNA binding activity, and nuclear translocation of Stat6. The Journal of Allergy and Clinical Immunology. doi:10.1016/s0091-6749(03)81014-x
• Daines, M. O., Hershey, G. K., Cunningham, C. M., Ericksen, M. B., & Andrews, R. P. (2003). Cycling of Stat6 is required for maintenance of Stat6 activation and IL-4 dependent gene induction. The Journal of Allergy and Clinical Immunology. doi:10.1016/s0091-6749(03)81301-5
• Daines, M. O., Hershey, G. K., Cunningham, C., Ericksen, M. B., & Andrews, R. J. (2002). Analysis of the Life Cycle of Stat6. Journal of Biological Chemistry. doi:10.1074/jbc.m200986200
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  Signal transducer and activator of transcription (Stat)6 is a transcription factor important for the development of Th2 cells and regulation of gene expression by IL-4 and IL-13. It is known that Stat6 is rapidly activated in response to IL-4; however, the fate of activated Stat6 is less clear. We examined the fate of activated Stat6 and found that during continuous exposure to IL-4, Stat6 activity was sustained for 72 h and that the maintenance of a constant level of activated Stat6 did not require new protein synthesis. In contrast, when cells were pulsed with IL-4 and then incubated in the absence of IL-4, the half-life of Stat6 phosphorylation and DNA binding activity was less than 1 h. Stat6 did not accumulate in the nucleus, and protein degradation did not play a major role in the disappearance of activated Stat6. Inhibition of kinase activity by staurosporine or the JAK inhibitor, AG490, revealed that maintenance of Stat6 activation in the continuous presence of IL-4 required ongoing phosphorylation of latent cytoplasmic Stat6 molecules. Cells treated with an inhibitor of nuclear export, leptomycin B, were unable to maintain Stat6 activation. Thus, the maintenance of Stat6 activation requires a constant cycle of activation, deactivation, nuclear export, and reactivation.
• Hershey, G. K., & Daines, M. O. (2002). Phosphorylated Stat-6 in cytoplasm is devoid of DNA binding activity. The Journal of Allergy and Clinical Immunology, 109(1), S361-S361. doi:10.1016/s0091-6749(02)82260-6

Proceedings Publications

• Pereira, R. S., Daines, M. O., & Cunningham, A. (2012). Differential Immune Responses To Alternaria Alternata Components. In D21. MICROBIAL CONTRIBUTIONS TO ASTHMA PATHOGENESIS.
• Pereira, R. S., Daines, M. O., & Cunningham, A. (2012). Impact Of Early Postnatal Exposure To Alternaria On Mouse Lung Development. In A55. LUNG DEVELOPMENT AND CHRONIC LUNG DISEASE OF PREMATURITY.
• Daines, M. O., Shultz, S., Sherwood, C. L., Hoffman, J., Gruzinova, I., Flynn, A. N., Daines, M. O., & Boitano, S. (2011). Alternaria Alternata Serine Proteases Induce Lung Inflammation And Airway Epithelial Cell Activation Via Protease-Activated Receptor-2 (PAR-2). In A107. NEW CONCEPTS IN PROTEASES AND ANTIPROTEASES.

Presentations

• Kiela, P. R., West, J. J., Murray, M., Galindo, K., Mayate-Ortiz, L., Laubitz, D., Daines, M. O., & Rice, S. A. (2019, May). Assessing the Causes, Epidemiology and Under-diagnosis of Pediatric Acute-onset Neuropsychiatric Syndrome (PANS) in Arizonans. 4th Annual ABRC Research Conference, Phoenix, AZ May 2, 2019. Phoenix, AZ: Arizona Biomedical Research Commission.

Poster Presentations

• Eremija, J., & Daines, M. O. (2023, spring). Neuropsychiatric Testing Provides Objective Insight Into Beneficial Effects Of Intravenous Immunoglobulins In Patients With Pediatric Acute-Onset Neuropsychiatric Syndrome. AAAAI.
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  poster presentation
• Mayate, L., Kiela, P. R., Daines, M. O., & Rice, S. A. (2021, February). Assessing the Causes, Epidemiology and Under-diagnosis of Pediatric Acute-onset Neuropsychiatric Syndrome (PANS) in Arizonans. Arizona Biomedical Research Commission, Annual Research Conference. Phoenix, Arizona: ABRC.
• Buckley, R. D., Patel, S., & Daines, M. O. (2017, March). Bacterial meningitis in pediatric patient with C2 deficiency. AAAAI 2017 annual meeting. Atlanta, GA: AAAAI/Journal of Allergy an Clinical Immunology.
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  Case presentation of a patient with complement C2 deficiency and normal antibody levels that had bacterial meningitis after stopping IVIG treatment
• Patel, s., Carr, T. F., & Daines, M. O. (2017, March). ORAI1 Mutation in a Child with Primary Immunodeficiency-9. AAAAI 2017 national meeting/Journal of Allergy and Clinical Immunology Supplement. Atlanta, GA: AAAAI.
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  Case presentation and discussion of pathology, diagnosis, and treatment of immune deficiency 9
• Mathur, A., Stern, D., Daines, M. O., Wright, A. L., Martinez, F., & Carr, T. F. (2016, February). Sensitivity and Specificity of a Clinical Diagnosis of Allergic Rhinitis in Childhood. AAAAI 2016 annual meeting. Los Angeles, CA: AAAAI/Journal of Allergy and Clinical Immunology.
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  We sought to evaluate the sensitivity and specificity of a clinical diagnosis of allergic rhinitis, using history and physical, to identify skin test positive rhinitis.