Citations 163
Reactivity of monoclonal antibodies EG1 and EG2 with eosinophils and their granule proteins
Nakajima, Hirokazu, Loegering, David A., Kita, Hirohito , Kephart, Gail M., Gleich, Gerald J. , Kita, H
Journal of Leukocyte Biology | January 12, 2018 | Volume: 66 | Issue: 3 | Pages: 447-454
Abstract Use of the murine monoclonal antibodies EG1 and EG2 has been based on the assumption that EG2 recognizes activated eosinophils. We examined the reactivity of EG1 and EG2 with eosinophil cationic protein (ECP), eosinophil-derived neurotoxin (EDN), and stimulated and nonstimulated eosinophils from normal donors. By radioimmunoassay, EG1 recognized only ECP, whereas EG2 recognized both ECP and EDN. By Western blot, EG1 reacted with ECP, EG2 reacted with both ECP and EDN, but EG2 could not distinguish between lysates of stimulated and nonstimulated eosinophils. By immunofluorescence, EG1 and EG2 at 20 μg/mL stained 95–100% of nonstimulated eosinophils, regardless of fixative; EG1 and EG2 at 0.1 μg/mL stained 61–90% of acetone- and paraformaldehyde-fixed and only 5–21% of methanol-fixed nonstimulated eosinophils. Thus, the reactivity of EG1 and EG2 with eosinophils depends on the method of fixation and antibody concentration; and EG2, in contrast to previous reports, cannot reliably discriminate between resting and activated eosinophils. J. Leukoc. Biol. 66: 447–454; 1999.
Eosinophil granule proteins in peripheral blood granulocytes
Abu-Ghazaleh, R I, Dunnette, Sandra L., Loegering, David A., Checked, James L, Kita, Hirohito , Thomas, Larry L., Gleich, Gerald J. , Kita, H
Journal of Leukocyte Biology | January 12, 2018 | Volume: 52 | Issue: 6 | Pages: 611-618
Abstract Eosinophils contain four principal cationic proteins, major basic protein (MBP), eosinophil-derived neurotoxin (EDN), eosinophil cationic protein (ECP), and eosinophil peroxidase (EPO). To determine the quantities of these proteins in granulocytes and whether they are specific to eosinophils, their concentrations in ly- sates of human granulocytes were measured using specific radioimmunoassays. The effect of different methods for eosinophil lysis on the recovery of the proteins was also studied. Maximal recovery occurred at pH 2 for MBP and pH 5.6 for the other granule proteins. The proteins cosedimented with eosinophils and their concentrations (X ± SEM) in ng/106 eosinophils (and in nM/106 eosinophils) were: MBP, 8,982 ± 611 (641.6); EDN, 3,283 ± 116 (178.4); ECP, 5,269 ± 283 (250.9); and EPO, 12,174 ± 859 (171.5). Basophils from a normal person contained (in ng/106 cells) MBP, 2,374; EDN, 214; ECP, 77; and EPO, 17. Highly purified neutrophils contained (in ng/106 cells) MBP, 3 ± 0.5; EDN, 72 ± 9; and ECP, 50 ± 12. Therefore we conclude that these proteins are mainly expressed in eosinophils, but that certain ones are present in basophils and neutrophils.
Localization of eosinophil-derived neurotoxin and eosinophil cationic protein in neutrophilic leukocytes
Sur, Sanjiv , Glitz, Dohn G., Kita, Hirohito , Kujawa, Stephen M., Peterson, Ellen A., Weiler, Deborah A., Kephart, Gail M., Wagner, Jill M., George, Terry J., Gleich, Gerald J. , Leiferman, Kristin M. , Kita, H
Journal of Leukocyte Biology | January 12, 2018 | Volume: 63 | Issue: 6 | Pages: 715-722
Abstract Eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) are generally regarded as eosinophil-specific proteins. We tested whether EDN and ECP are present in mature neutrophils. By indirect immunofluorescence, both eosinophils and neutrophils stained with antibodies to EDN and ECP. Lysates of purified (<0.1% eosinophil contamination) neutrophils contained EDN, 112 ± 4 ng/106 cells, and ECP, 163 ± 2 ng/106 cells, whereas eosinophil major basic protein (MBP) was not detectable. Electron microscopic examination of immunogold-labeled buffy coat cells stained with EDN antibody showed that EDN is localized to neutrophil granules. Finally, EDN mRNA was detected in lysates of highly purified neutrophils (0.001% eosinophil contamination) by the reverse transcription-polymerase chain reaction. We conclude that proteins that are either identical to or immunologically cross-reactive with EDN and ECP are present in neutrophils and that EDN is synthesized and localized to neutrophil granules. Thus, caution must be exercised in interpreting the presence of EDN and ECP as specific markers of eosinophil-associated inflammation in human disease.
IL-33–Responsive Group 2 Innate Lymphoid Cells Are Regulated by Female Sex Hormones in the Uterus
Bartemes, Kathleen R. , Iijima, Koji, Drake, Li Y. , Kita, Hirohito , Chen, Chien-Chang, Drake, Li , Kita, H , Chen, Chien‐Chang
The Journal of Immunology | November 13, 2017 | Volume: 200 | Issue: 1 | Pages: 229-236
Abstract Group 2 innate lymphoid cells (ILC2s) reside in multiple organs in the body, where they play roles in immunity, tissue homeostasis, and metabolic regulation. However, little is known about the regulatory mechanisms of ILC2s in different organs. Here, we identified ILC2s in the mouse uterus and found that they express cell surface molecules, including the IL-33 receptor, ST2, that are roughly comparable to those expressed by lung ILC2s. Both in vivo and in vitro treatment with IL-33 induced type 2 cytokine production in uterine ILC2s, suggesting that they respond to IL-33 in a manner similar to ILC2s in other organs. Importantly, uterine ILC2s were nearly absent in ovariectomized mice and were increased in wild-type mice by estrogen administration, whereas lung ILC2s were unaffected by both ovariectomy and estrogen administration. Likewise, a marked reduction in uterine ILC2s was observed in mice deficient in estrogen receptor α or estrogen receptor β. Furthermore, uterine ILC2s highly expressed estrogen receptor α, and in vitro culture of isolated uterine ILC2s with 17β-estradiol modified expression of a number of genes. Finally, an increased prevalence in neonatal mortality was observed in litters from dams lacking the IL-33 receptor, ST2. Taken together, our findings indicate that unlike lung IL2Cs, uterine ILC2s are regulated by female sex hormones, which may specialize them for specific physiological functions.
CELLULAR SENESCENCE DRIVES FIBROTIC PULMONARY DISEASE
White, Thomas A. , Iijima, Koji, Mazula, Daniel L., Passos, JF, Kirkland, James L. , Schafer, Marissa , Kita, Hirohito , LeBrasseur, Nathan K.
Innovation in Aging | July 1, 2017 | Volume: 1 | Issue: suppl_1 | Pages: 135-135
IL ‐33: biological properties, functions, and roles in airway disease
Kita, Hirohito , Drake, Li Yin, Kita, H , Drake, Li Y.
Immunological Reviews | June 28, 2017 | Volume: 278 | Issue: 1 | Pages: 173-184
Summary Interleukin ( IL )‐33 is a key cytokine involved in type 2 immunity and allergic airway diseases. Abundantly expressed in lung epithelial cells, IL ‐33 plays critical roles in both innate and adaptive immune responses in mucosal organs. In innate immunity, IL ‐33 and group 2 innate lymphoid cells ( ILC 2s) provide an essential axis for rapid immune responses and tissue homeostasis. In adaptive immunity, IL ‐33 interacts with dendritic cells, Th2 cells, follicular T cells, and regulatory T cells, where IL ‐33 influences the development of chronic airway inflammation and tissue remodeling. The clinical findings that both the IL ‐33 and ILC 2 levels are elevated in patients with allergic airway diseases suggest that IL ‐33 plays an important role in the pathogenesis of these diseases. IL ‐33 and ILC 2 may also serve as biomarkers for disease classification and to monitor the progression of diseases. In this article, we reviewed the current knowledge of the biology of IL ‐33 and discussed the roles of the IL ‐33 in regulating airway immune responses and allergic airway diseases.
Airway epithelial anion secretion and barrier function following exposure to fungal aeroallergens: role of oxidative stress
Zaidman, Nathan A. , O’Grady, Kelly E., Patil, Nandadevi, Milavetz, Francesca, Maniak, Peter J., Kita, Hirohito , O’Grady, Scott M. , Kita, H
American Journal of Physiology-Cell Physiology | April 27, 2017 | Volume: 313 | Issue: 1 | Pages: C68-C79
Aeroallergens produced by Alternaria alternata can elicit life-threatening exacerbations of asthma in patients sensitized to this fungus. In this study, the effect of Alternaria on ion transport mechanisms underlying mucociliary clearance and airway epithelial barrier function was investigated in human airway epithelial cells. Apical exposure to Alternaria induced an increase in anion secretion that was inhibited by blockers of CFTR and Ca2+-activated Cl− channels. Stimulation of anion secretion was dependent on Ca2+ uptake from the apical solution. Alternaria exposure also produced an increase in reactive oxygen species (ROS) that was blocked by pretreatment with the oxidant scavenger glutathione (GSH). GSH and the NADPH oxidase inhibitor/complex 1 electron transport inhibitor diphenylene iodonium chloride (DPI) blocked ATP release and the increase in intracellular [Ca2+] evoked by Alternaria. Alternaria also decreased transepithelial resistance, and a portion of this effect was dependent on the increase in ROS. However, the Alternaria-induced increase in unidirectional dextran (molecular mass = 4,000 Da) flux across the epithelium could not be accounted for by increased oxidative stress. These results support the conclusion that oxidative stress induced by Alternaria was responsible for regulating Ca2+-dependent anion secretion and tight junction electrical resistance that would be expected to affect mucociliary clearance.
Correlation of Symptoms, Clinical Signs, and Biomarkers of Inflammation in Postsurgical Chronic Rhinosinusitis
Racette, Samuel D. , Wijewickrama, Rohan C., Jayaprakash, Vijayvel , Sherris, David A., Kita, Hirohito , O’Donnell, Frank E., Ponikau, Jens U., Santos, Carlos Henrique Marques dos , Kita, H
Annals of Otology, Rhinology & Laryngology | April 5, 2017 | Volume: 126 | Issue: 6 | Pages: 455-462
Objective: The study aimed to evaluate symptoms described by patients with chronic rhinosinusitis with polypoid changes/nasal polyps and their correlation with computed tomography (CT), nasal endoscopy, and intranasal biomarkers. Study Design: Prospective multicenter study symptom data from postsurgical adult chronic rhinosinusitis study participants with recurrent disease refractory to medical therapy were analyzed in comparison with objective data. Methods: Using logistic regression analysis, participant-rated 16-question surveys from 258 participants were assessed for correlation with nasal endoscopy scores, CT percentage of sinus occlusion, and intranasal biomarkers of fungal antigens (Alternaria and Aspergillus), eosinophilic inflammation (eosinophil-derived neurotoxin [EDN] and major basic protein [MBP]), and inflammatory cytokines (interleukins 5 and 13). Results: Study participant assessments revealed increased CT occlusion in participants presenting with greater inability to smell ( P < .019). Mucosal inflammation identified on nasal endoscopy was positively correlated with congestion ( P < .028), runny nose ( P < .002), and ear pain ( P < .007). Elevated EDN was positively correlated in patients with bothersome congestion ( P < .031) and runny nose ( P < .011). Sneezing was positively correlated with multiple markers: Alternaria ( P < .024), interleukin-13 ( P < .027), MBP ( P < .034), and interleukin-5 ( P < .019). Conclusion: Nasal endoscopy, not CT imaging, has the strongest correlation with the 2 cardinal symptoms of congestion and runny nose in CRS patients; these correlate with biomarkers of eosinophilic inflammation.
Application of a Natural Language Processing Algorithm to Asthma Ascertainment. An Automated Chart Review
Sohn, Sunghwan , Rolfes, Mary , Seabright, Alicia, Ryu, Euijung , Voge, Gretchen A., Bachman, Kay, Park, Miguel A., Kita, Hirohito , Croghan, Ivana T. , Liu, Hongfang , Juhn, Young J. , Bachman, Kay A, Wi, Chung‐Il , Kita, H
American Journal of Respiratory and Critical Care Medicine | April 4, 2017 | Volume: 196 | Issue: 4 | Pages: 430-437
Abstract Rationale Difficulty of asthma ascertainment and its associated methodologic heterogeneity have created significant barriers to asthma care and research. Objectives We evaluated the validity of an existing natural language processing (NLP) algorithm for asthma criteria to enable an automated chart review using electronic medical records (EMRs). Methods The study was designed as a retrospective birth cohort study using a random sample of 500 subjects from the 1997–2007 Mayo Birth Cohort who were born at Mayo Clinic and enrolled in primary pediatric care at Mayo Clinic Rochester. Performance of NLP-based asthma ascertainment using predetermined asthma criteria was assessed by determining both criterion validity (chart review of EMRs by abstractor as a gold standard) and construct validity (association with known risk factors for asthma, such as allergic rhinitis). Measurements and Main Results After excluding three subjects whose respiratory symptoms could be attributed to other conditions (e.g., tracheomalacia), among the remaining eligible 497 subjects, 51% were male, 77% white persons, and the median age at last follow-up date was 11.5 years. The asthma prevalence was 31% in the study cohort. Sensitivity, specificity, positive predictive value, and negative predictive value for NLP algorithm in predicting asthma status were 97%, 95%, 90%, and 98%, respectively. The risk factors for asthma (e.g., allergic rhinitis) that were identified either by NLP or the abstractor were the same. Conclusions Asthma ascertainment through NLP should be considered in the era of EMRs because it can enable large-scale clinical studies in a more time-efficient manner and improve the recognition and care of childhood asthma in practice.
Kita, Hirohito
Nihon Shoni Arerugi Gakkaishi. The Japanese Journal of Pediatric Allergy and Clinical Immunology | March 18, 2017 | Volume: 14 | Issue: 3 | Pages: 303-303