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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。
To investigate the proliferation and differentiation of CD4 ~ + CD25 ~ + Foxp3 ~ + Tregs and the secretion of inhibitory cytokines TGF-β1 and IL-10 in blood of children with different pathogen of bronchiolitis (hereinafter referred to as: Impact. The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Tregs in children with rheumatoid arthritis was measured by using RSV and non-RSV infection. Mononuclear cells (PBMCs) isolated from the blood were inoculated into 24-well plates And the cells were blocked with CD40L McAb. The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg in the culture plate was detected by flow cytometry after 72 hours. The surface of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells The average density of Foxp3 and the content of TGF-β1 and IL-10 in culture supernatants were detected by enzyme-linked immunosorbent assay. The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg in children with RSV hairy dermatitis was significantly lower than that in non-RSV hairy dermatitis children and normal controls (P <0.05) ~ + CD25 ~ + Foxp3 ~ + Treg percentage were significantly lower than non-RSV hairy dermatitis children, normal control group and anti-CD40L McAb group (P <0.05). The average density of Foxp3 on the surface of CD4 ~ + CD25 ~ + Foxp3 ~ + Tregs in PBMCs of anti-CD40L McAb group was significantly higher than that of RSV bronchiolitis group (P <0.05) . The levels of IL-10 and TGF-β1 in PBMC culture supernatant were significantly higher in anti-CD40L McAb group than in non-RSV bronchiolitis group, and were significantly higher in non-RSV bronchiolitis group than in RSV bronchitis group (P <0.05). The number of CD4 ~ + CD25 ~ + Foxp3 ~ + Tregs in RSV hairy dermatitis patients was not enough. Anti-CD40L McAb blocking CD40 / CD40L pathway in vitro could promote the proliferation of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg proliferation.