RSV毛细支气管炎患儿血液CD4^+CD25^+Foxp3^+Treg与CD40/CD40L关系的研究  

Study on the relationship between blood CD4^+CD25^+Foxp3^+ Treg and CD40/CD40L in children with RSV induced brochiolitis

作　　者：石涛[1] 吴福玲[1] 李宾[2] 

机构地区：[1]滨州医学院附属医院儿科,滨州256603 [2]滨州医学院儿科学教研室

出　　处：《现代免疫学》2012年第5期425-429,共5页Current Immunology

基　　金：山东省自然科学基金资助项目(ZR2009CL007)

摘　　要：探讨CD40/CD40L信号通路对不同病原体毛细支气管炎(以下简称:毛支炎)患儿血液CD4^+CD25^+Foxp3^+ Treg的增殖分化及其分泌抑制性细胞因子TGF-β1、IL-10的影响。取呼吸道合胞病毒(RSV)及非RSV感染毛支炎患儿血液并检测CD4^+CD25^+Foxp3^+Treg的百分率;并将血液中分离的单个核细胞(PBMC)接种于24孔板内,加入CD40L McAb进行阻断,作用72h后采用流式细胞仪检测培养板内CD4^+CD25^+Foxp3^+Treg的百分率,激光共聚焦检测CD4^+CD25^+Foxp3^+Treg细胞表面Foxp3的平均密度,酶联免疫吸附法检测培养上清中TGF-β1、IL-10的含量。RSV毛支炎患儿血液中CD4^+CD25^+Foxp3^+Treg百分率显著低于非RSV毛支炎患儿及正常对照组(P<0.05);RSV毛支炎患儿体外培养PBMC中CD4^+CD25^+Foxp3^+Treg百分率均显著低于非RSV毛支炎患儿、正常对照组和抗CD40L McAb组(P<0.05)。毛支炎患儿PBMC经过体外培养72h后,抗CD40L McAb组培养的细胞中CD4^+CD25^+Foxp3^+Treg表面Foxp3的平均密度显著高于RSV毛支炎患儿组(P<0.05)。PBMC培养上清中IL-10和TGF-β1的水平抗CD40L McAb组明显高于非RSV毛支炎组,非RSV毛支炎组明显高于RSV毛支炎组,差异有统计学意义(P<0.05)。RSV毛支炎患儿体内存在严重的CD4^+CD25^+Foxp3^+Treg数量不足;体外用抗CD40L McAb阻断CD40/CD40L通路可促进RSV毛支炎PBMC中CD4^+CD25^+Foxp3^+Treg的增殖。Abstract： To determine the effect of the CD40/CD40L signaling pathway to the proliferation and differentiation of CD4 ＋ CD25＋ Foxp3＋ Treg and the secretion of inhibitory cytokines, TGFβ1, IL10 in the blood of the bronchiolitis infected with different pathogens, the blood of the children who developed bronchiolitis which infected or uninfected with RSV were collected and the percentage of the CD4＋ CD25＋ Foxp3＋ Treg was detected by FCM（flow cytometer）. Lymphocytes were isolated and seeded in 24 well plate, after 72hcoculture with CD40L monoclonal antibody to block, the percentage of the CD4＋ CD25＋ Foxp3＋ Treg were detected by FCM（flow cytometer） ,the expression of PECY5Foxp3 was detected by laser scanning cofocal microscopy, the levels of the TGF131and IL10 in supernatant were detected by enzymelinked immunesorbent assay. The percentage of CD4＋CD25＋ Foxp3＋ Treg in blood of the children who were infected with RSV and developed bronchiolitis （RSV infected group） was significantly lower than those in the RSV uninfected group and the control group （P〈0.05）. In the in vitro culture system, the percentage of CD4＋ CD25＋ Foxp3＋ Treg in RSV infected group was significantly lower than in RSV uninfected group, the control group and the anti CD40L McAb group（P〈0.05）. Cocuhured with the CD40L monoclonal antibody, 72h later, the average optical density of the PE CYSFoxp3 in the surface of CD4＋ CD25＋ Foxp3＋ Treg in antiCD40L McAb group was higher than in the RSV infected group. （P〈0.05）. The levels of IL10 and TGF131 in the supernatant of the mixed lym phocyte in antiCD40L McAb group were significantly higher than in the RSV uninfected group. The RSV uninfected group were significantly higher than the RSV uninfected group. The number of CD4＋ CD25＋ Foxp3＋ Treg in the blood of the children who developed bronchiolitis due to RSV infection is lower than that in the normal controls. The in vitro study demonstrated that blocking the CD40/CD40L signaling with ant

关 键 词：毛细支气管炎 CD40/CD40L CD40L McAb TGF-β1 IL-10 

分 类 号：R392.12[医药卫生—免疫学]