In order to determine whether or not IFN-gamma R is associated with regulatory mechanisms on human eosinophil function, we examined the expression of functional IFN-gamma R on human peripheral eosinophils. In this study, peripheral blood eosinophils were obtained from seven normal controls and 12 patients (bronchial asthma, n = 9, and hypereosinophilic syndrome (HES), n = 3), and the purity of eosinophils was 97.11 +/- 2.31%, n = 19. We first showed that anti-IFN-gamma R alpha-chain MoAb reacted with all tested eosinophils of both normal controls and patients by flow cytometry analysis. We also showed expression of mRNA for the alpha-chain of IFN-gamma R in all purified eosinophils of six individuals. Further, to characterize IFN-gamma R on eosinophils, we did binding experiments with I-125-IFN-gamma on purified peripheral eosinophils. The linear Scatchard plot indicated a single type of high-affinity binding sites (dissociation constant (Kd) = 3.89-4.95x10(-10) M, numbers of binding sites = 183-233/cell, n = 3). To determine whether IFN-gamma R on eosinophils is functional, we examined surface eosinophilic cationic protein (ECP) and CD69 induction after IFN-gamma R Ligation with recombinant human IFN-gamma (rhIFN-gamma) on eosinophils by flow cytometry. rhIFN-gamma stimulation significantly induced both ECP and CD69 expression on the 2-18 h-cultured eosinophils in a dose-dependent manner. Further, the effects of rhIFN-gamma stimulation were significantly blocked by both a neutralizing anti-IFN-gamma MoAb and a blocking anti-IFN-gamma R MoAb. These results suggest that human peripheral eosinophils express functional IFN-gamma R.
机构:
SUNY Stony Brook, Div Pulm Crit Care & Sleep Med, Stony Brook, NY 11794 USASUNY Stony Brook, Div Pulm Crit Care & Sleep Med, Stony Brook, NY 11794 USA
Fusiak, Timothy
Smaldone, Gerald C.
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SUNY Stony Brook, Div Pulm Crit Care & Sleep Med, Stony Brook, NY 11794 USASUNY Stony Brook, Div Pulm Crit Care & Sleep Med, Stony Brook, NY 11794 USA
Smaldone, Gerald C.
Condos, Rany
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NYU, Sch Med, Div Pulm & Crit Care Med, New York, NY USASUNY Stony Brook, Div Pulm Crit Care & Sleep Med, Stony Brook, NY 11794 USA
机构:
Shandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R ChinaShandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R China
Ding, Huihui
Wang, Gongfu
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China Food & Drug Adm CFDA, Ctr Drug Evaluat, Beijing, Peoples R ChinaShandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R China
Wang, Gongfu
Yu, Zhen
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Shandong First Med Univ, Dept Pharm, Cent Hosp, Jinan, Shandong, Peoples R ChinaShandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R China
Yu, Zhen
Sun, Huimin
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Shandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R ChinaShandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R China
Sun, Huimin
Wang, Lu
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Shandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R China
Shandong First Med Univ, Dept Pharm, Cent Hosp, Jinan, Shandong, Peoples R ChinaShandong First Med Univ, Sch Pharm, Jinan, Shandong, Peoples R China