人T细胞杂交瘤组成性分泌的B细胞生长因子的部分纯化及特性分析

Partial purification and characterization of B cell growth factor constitutively secreted by human T-cell hybridoma.

作者信息

Pathak S K, Tsang K Y, Cathcart M K, Arnaud P, Boutin B, Fudenberg H H

出版信息

Immunol Lett. 1985;10(6):339-46. doi: 10.1016/0165-2478(85)90129-4.

DOI:10.1016/0165-2478(85)90129-4

Abstract

Human T cell hybridomas were established by fusion of PHA-activated PBL with the 8-azaguanine resistant human T-leukemic cell line CEM-CM3. High levels of B cell growth factor (BCGF) activity were detected in the supernatants of hybridoma C8-2B2 and its subclones. Hybridoma C8-2B2, in addition to the Leu 3a, also expressed the OKT11 surface marker which was not detectable on the parent CEM-CM3 cells. BCGF from the culture supernatant was purified by combined use of salt fractionation and gel filtration to 36.6 fold with 23.9% recovery of activity. The BCGF produced by hybridoma C8-2B2 has a molecular weight range of 16,000-20,000 in two major electrophoretically different forms with pI values of 6.4 and 7.4.

摘要

通过将PHA激活的外周血淋巴细胞（PBL）与8-氮杂鸟嘌呤抗性人T白血病细胞系CEM-CM3融合，建立了人T细胞杂交瘤。在杂交瘤C8-2B2及其亚克隆的上清液中检测到高水平的B细胞生长因子（BCGF）活性。杂交瘤C8-2B2除了表达Leu 3a外，还表达了OKT11表面标志物，而在亲本CEM-CM3细胞上未检测到该标志物。通过盐析和凝胶过滤相结合的方法，从培养上清液中纯化BCGF，纯化倍数达到36.6倍，活性回收率为23.9%。杂交瘤C8-2B2产生的BCGF分子量范围为16,000 - 20,000，有两种主要的电泳不同形式，其pI值分别为6.4和7.4。

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