DEVELOPMENT OF A HUMAN T-CELL HYBRIDOMA SECRETING SEPARATE B-CELL GROWTH AND DIFFERENTIATION FACTORS
DEVELOPMENT OF A HUMAN T-CELL HYBRIDOMA SECRETING SEPARATE B-CELL GROWTH AND DIFFERENTIATION FACTORS
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DOI：
10.1073/pnas.81.8.2475
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
FAUCI, AS
中科院分区：
文献类型：
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作者：
BUTLER, JL;FALKOFF, RJM;FAUCI, AS
建立了一株分泌B细胞生长因子（BCGF）和B细胞分化因子（BCDF）的克隆化人T细胞杂交瘤（7 D5）。来自该杂交体的上清液能够在抗IgM激活的正常人B细胞中维持增殖。杂交瘤上清诱导金黄色葡萄球菌科万I刺激的B细胞分化和抗体分泌。该杂交瘤的上清液中不存在白细胞介素2。通过凝胶过滤色谱法测定来自BCGF和BCDF的杂交瘤的分子大小。BCGF活性存在于20-kDa [千道尔顿]级分中，而BCDF活性洗脱于30-至35-kDa级分中。用色谱聚焦法测得各因子的等电点分别为BCGF 6.6和BCDF 5.9。使用特定靶细胞进行吸收实验。植物血凝素刺激的T细胞母细胞没有去除BCGF或BCDF活性。抗IgM激活的B细胞吸收BCGF，但不吸收BCDF。CESS [人B淋巴母细胞]细胞去除BCDF，但不去除BCGF。因此，开发了分泌2种不同B细胞淋巴因子的人T细胞杂交瘤。这些免疫调节分子的进一步免疫化学和功能研究应大大提高对正常和疾病状态下人类B细胞功能调节的理解。
A cloned human T-cell hybridoma (7D5), secreting B-cell growth factor (BCGF) and B-cell differentiation factor (BCDF), was established. Supernatant from this hybrid was capable of maintaining proliferation in anti-IgM-activated normal human B cells. The hybridoma supernatant induced differentiation and antibody secretion in Staphylococcus aureus Cowan I-stimulated B cells. No interleukin 2 was present in supernatant from this hybridoma. Molecular size of the hybridoma-derived from BCGF and BCDF was determined by gel filtration chromatography. BCGF activity was present in the 20-kDa [kiloDalton] fractions, and BCDF activity eluted in the 30- to 35-kDa fractions. The isoelectric points of the factors, determined by chromatofocusing, were 6.6 for BCGF and 5.9 for BCDF. Absorption experiments were performed using specific target cells. Phytohemagglutinin-stimulated T-cell blasts did not remove either BCGF or BCDF activity. Anti-IgM-activated B cells absorbed BCGF but not BCDF. CESS [human B-lymphoblastoid cells] cells removed BCDF but not BCGF. Thus, a human T-cell hybridoma secreting 2 distinct B-cell lymphokines was developed. Further immunochemical and functional studies of these immunoregulatory molecules should greatly enhance understanding of the regulation of human B-cell function in normal and disease states.