分泌白细胞介素-2T细胞杂交瘤株的建立

Establishment of a T Hybridoma Secreting Interleukin 2

本研究利用T淋巴细胞杂交瘤技术建立了能持续稳定地分泌白细胞介素—2(IL—2)的小鼠T淋巴细胞杂交瘤株,首先用刀豆蛋白A(ConA)活化BALB/C小鼠的脾细胞,然后使其与小鼠胸腺瘤细胞BW5147融合,用四甲基偶氮唑盐(MTT)比色分析法从261个杂交克隆中筛选出至少30个分泌IL—2的T细胞杂交瘤株,对选出的一个杂交瘤细胞株FB4作了进一步研究,该杂交瘤株的染色体平均44(42～47)条,经传代培养、冻存和复苏后仍能持续稳定地分泌IL—2,用含2％小牛血清的DMEM完全培养基大量培养FB4杂交瘤细胞,制得含IL—2的培养上清,再经沉淀和凝胶过滤等步骤制得了部分纯化的IL—2,用十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS—PAGE)技术检出了IL—2成分,其表观分子量(MW)约28KDa。

A T hybridoma cell line secreting interleukin 2 (IL-2) was established by lymphocyte hy-bridoma techniques. Spleocytes from BALB /C mouse were activated by concanavalin A (Con A) and then fused with BW5417 T cell lymphoma. At least 30 T hybridomas secreting IL-2 were selected from 261 hybrid clones by MTT colorimetric assay and of which,a hybridoma FB4 was cloned and used in following studies. This hybridoma,with 44 (42-47) chromosomes,can maintain to produce IL-2 stably after long-term propagating,freezing and thawing. The IL-2 was partially purified from the FB4 hybridoma culture supernatant by sulphate precipitation and gel chromatograph. On the pattern of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) ,the IL-2 was detected with apparent molecular weight(MW)of 28KDa.