抗hGITR_(aa27-165)多克隆抗体生物学活性的初步鉴定  

Primary detection of biological functions of anti-hGITR_(aa27-165)PcAb

作　　者：蔡玲[1] 刘艳[2] 张慧[3] 韩清珍[2] 郑东[2] 陈建国[1] 

机构地区：[1]江苏大学附属人民医院检验科,江苏镇江212002 [2]江苏大学检验医学研究所,江苏镇江212013 [3]江苏大学附属医院检验科,江苏镇江212001

出　　处：《细胞与分子免疫学杂志》2011年第11期1217-1219,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省高校自然科学基础研究项目(08KJB320002);江苏省卫生厅面上项目(H200859)

摘　　要：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。AIM： To detect the rabbit-derived polyclonal antibodies against extracellular protein segments of human glucocorticoid-inducible tumor necrosis factor receptors （anti-hGITRaa27- 165 PcAb） with regard to its capacity of linkage to natural GITR molecules and the function on CD4＋ T cells, METHODS： Human peripheral blood mononuclear cells （PBMCs） were isolated and cultured under stimulation; flow cytometry was applied to check the capacity of anti- hGITRaa27- 165 PcAb for linkage to natural GITR molecules on PBMCs; human CD4 ＋ T cells were isolated by immunological magnetic beads and 3H-TdR incorporation tests were performed to observe improving-proliferation action of anti- hGITRaa27- 165 PcAb while CD4＋ T cells were cultured with or not with some cytokines. RESULTS： Anti-hGITRaa27- 165 PcAb was able to bind GITR molecules with natural conformation in a concentration-dependent way; furthermore, this PcAb could improve the reproduction of CD4＋ T cells. CONCLUSION： The rabbit-derived anti-hGITRaa27- 165PcAb prepared in our laboratory is capable of linking to natural target molecules and possesses the activation function upon CD4＋ T cells, the further exploration should allow for its applications for diagnosis and treatment of relavent diseases.

关 键 词：糖皮质激素诱导的肿瘤坏死因子受体 多克隆抗体 流式细胞术 CD4+T细胞 

分 类 号：R394[医药卫生—医学遗传学]