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检测TNF受体胞外区在RBL-2H3细胞中表达及分泌后的生物活性。用Ⅰ型TNF受体(TNFR1)胞外区的表达载体转染RBL-2H3细胞,用免疫荧光技术检测TNFR1的亚细胞定位,通过诱导IgE受体交联诱导细胞脱粒,用免疫沉淀结合SDS-PAGE检测TNFR1,用TNF-α敏感细胞株WEHI细胞检测分泌的TNFR1的生物活性。免疫荧光结果显示RBL-2H3细胞的分泌型溶酶体中存在TNFR1,免疫沉淀结果显示TNFR1在RBL-2H3细胞脱粒中释放,对WEHI细胞的TNF-α毒性实验显示分泌的TNFR1具有降低TNF-α细胞毒的作用。提示在RBL-2H3细胞中表达及分泌的TNFR1具有结合TNF-α的生物活性,能够降低TNF-α的细胞毒作用。
The biological activity of the TNF receptor extracellular domain was expressed and secreted in RBL-2H3 cells. RBL-2H3 cells were transfected with the expression vector of extracellular domain of type I TNF receptor (TNFR1), the subcellular localization of TNFR1 was detected by immunofluorescence technique, and cell degranulation was induced by inducing IgE receptor cross-linking. Immunoprecipitation and SDS- PAGE was used to detect TNFR1. The biological activity of secreted TNFR1 was detected by TNF-α sensitive cell line WEHI cells. Immunofluorescence results showed that TNFR1 was present in secretory lysosomes of RBL-2H3 cells. Immunoprecipitation results showed that TNFR1 was released from the degranulation of RBL-2H3 cells. Toxicity of TNF-α to WEHI cells showed that TNFR1 secretion decreased TNF- Cytotoxic effects. It is suggested that TNFR1 expressed and secreted in RBL-2H3 cells has the biological activity of binding to TNF-α, which can reduce the cytotoxic effect of TNF-α.