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目的:明确兔源性抗人糖皮质激素诱导的肿瘤坏死因子受体胞外段的多克隆抗体(anti-hGITRaa27-165 PcAb)与人类天然GITR分子的结合能力及其对CD4+T细胞的生物学作用。方法:分离、刺激培养人外周血单个核细胞后,利用流式细胞术(FCM)检测anti-hGITRaa27-165 PcAb与细胞膜表面GITR分子的结合能力;免疫磁珠分离人外周血收集CD4+T细胞,以3H-TdR掺入试验分别检测在细胞刺激生长剂存在或不存在的情况下,anti-hGITRaa27-165PcAb对CD4+T细胞增殖的影响。结果:兔源性anti-hGITRaa27-165 PcAb可以与天然构象的GITR分子结合,且此种结合具有浓度依赖性;在细胞生长刺激因子存在或不存在情况下,该多抗均能促进CD4+T细胞增殖。结论:本室制备的兔源性anti-hGITRaa27-165 PcAb具有与天然分子结合的能力以及一定的生物学活性,进一步的研究可考虑将其应用于临床相关疾病的检测和治疗。
OBJECTIVE: To determine the binding ability of anti-hGITRaa27-165 PcAb, an extracellular fragment of tumor necrosis factor receptor induced by rabbit anti-human glucocorticoid, to human natural GITR molecule and its biological activity on CD4 + T cells Learning role. METHODS: After culturing human peripheral blood mononuclear cells, the binding ability of anti-hGITRaa27-165 PcAb to GITR on cell membrane surface was detected by flow cytometry (FCM). CD4 + T cells were isolated from peripheral blood by immunomagnetic beads The effects of anti-hGITRaa27-165PcAb on the proliferation of CD4 + T cells were examined by 3H-TdR incorporation assay in the presence or absence of cell-stimulating agents. RESULTS: Rabbit-derived anti-hGITRaa27-165 PcAb could bind to the native conformation GITR molecule in a concentration-dependent manner. In the presence or absence of cytokine stimulator, the polyclonal antibody promoted CD4 + T Cell Proliferation. CONCLUSION: The rabbit-derived anti-hGITRaa27-165 PcAb prepared in our laboratory possesses the ability to bind to natural molecules and has certain biological activity. Further study may be considered for the detection and treatment of clinically relevant diseases.