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目的用异硫氰酸荧光素(FITC)标记抗马干扰素-γ(IFN-γ)单克隆抗体,为建立马IFN-γ细胞内细胞因子染色方法提供实验材料。方法利用FITC标记纯化的抗马IFN-γ单克隆抗体(SB10),荧光抗体经系列稀释后,对马外周血单核细胞(PBMC)进行细胞内细胞因子IFN-γ单染色的流式细胞术检测,同时与商品化FITC标记的抗牛IFN-γ单克隆抗体(CC302)进行流式细胞术检测对比试验。结果FITC-CC302染色马PBMC,1μl荧光抗体染色106个细胞时,分泌IFN-γ的细胞频率为9.35%,而FITC-SB10经系列稀释染色,在用0.25μl荧光抗体染色106个细胞时,分泌IFN-γ的细胞频率为9.90%。结论所制备的FITC标记的抗马IFN-γ单克隆抗体,可为建立马细胞免疫学研究方法奠定物质基础,进而为建立监测马机体免疫状态和研究机体免疫机制提供技术平台。
OBJECTIVE: To use monoclonal antibody against gamma-interferon-gamma (IFN-γ) labeled with fluorescein isothiocyanate (FITC) to provide experimental materials for the establishment of cytokine staining in horse IFN-γ cells. Methods FITC-labeled purified anti-MAb IFN-γ monoclonal antibody (SB10) was used. Fluorescent antibody was serially diluted and flow cytometry (FCM) was performed on PBMC of human peripheral blood mononuclear cells (PBMCs) , And compared with the commercialized FITC-labeled anti-bovine IFN-γ monoclonal antibody (CC302) for flow cytometry. Results When FITC-CC302 staining PBMC and 106μl cells were stained with 1μl fluorescent antibody, the frequency of IFN-γ-secreting cells was 9.35%. FITC-SB10 was serially diluted and secreted when cells were stained with 0.25μl fluorescent antibody The cell frequency of IFN-γ was 9.90%. Conclusion The FITC-labeled monoclonal antibody against horse IFN-γ could lay a material foundation for establishment of immunological research on horse cells, and provide a technological platform for monitoring immune status of the body and immune mechanism.