PRODUCTION OF A MONOCLONAL-ANTIBODY USEFUL IN THE MOLECULAR CHARACTERIZATION OF MURINE T-CELL-REPLACING FACTOR B-CELL GROWTH FACTOR-II
PRODUCTION OF A MONOCLONAL-ANTIBODY USEFUL IN THE MOLECULAR CHARACTERIZATION OF MURINE T-CELL-REPLACING FACTOR B-CELL GROWTH FACTOR-II
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DOI：
10.1073/pnas.84.13.4581
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发表时间：
1987-07-01
影响因子：
11.1
通讯作者：
TAKATSU, K
中科院分区：
文献类型：
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作者：
HARADA, N;TAKAHASHI, T;TAKATSU, K
t细胞替代因子（t -cell- replacement factor， TRF）是活化的B细胞向免疫球蛋白分泌细胞最终分化所必需的t细胞衍生因子。先前的研究表明，小鼠t细胞杂交体（B151K12）产生的因子可以(i)诱导B细胞白血病细胞系BCL1分泌免疫球蛋白，并由二硝基苯引发的B细胞体外合成次生抗2,4-二硝基苯IgG （TRF活性），（ii）诱导BCL1细胞增殖[B细胞生长因子ii （BCGF-II）活性]。这两种活动似乎都与同一分子有关。在这里，我们报道了一种小鼠TRF单克隆抗体的产生。该单克隆抗体命名为TB13，能强烈抑制TRF和BCGF-II活性，并能吸收由B151k12和注射pSP6K-mTRF23质粒转录mRNA的爪蟾卵母细胞产生的TRF和BCGF-II活性分子。抑制作用与TB13和TRF浓度均呈线性关系。单克隆抗体TB13不抑制b细胞刺激因子1、白细胞介素1、白细胞介素2和白细胞介素3的活性。用TB13获得的免疫沉淀溶解样品中的TRF活性在NaDodSo4/PAGE后恢复，在Mr为46,000的蛋白质带对应的部分中。我们的研究结果表明，单克隆抗体TB13识别具有TRF和BCGF-II活性的分子。
T-cell-replacing factor (TRF) is a T-cell-derived factor required for terminal differentiation of activated B cells to immunoglobulin-secreting cells. Previous studies have shown that a murine T-cell hybrid (B151K12) produces factor(s) that (i) induce immunoglobulin secretion by the B-cell leukemia line BCL1 and secondary anti-2,4-dinitrophenyl IgG synthesis in vitro by dinitrophenyl-primed B cells (TRF activity) and (ii) cause proliferation of the BCL1 cells [B-cell growth factor II (BCGF-II) activity]. Both activities appear to be associated with the same molecule. Here we report the production of a monoclonal antibody to murine TRF. The monoclonal antibody, designated TB13, strongly inhibited both TRF and BCGF-II activities and absorbed TRF-as well as BCGF-II-active molecules produced by B151k12 and by Xenopus oocytes that has been injected with mRNA transcribed from plasmid pSP6K-mTRF23. Inhibition was linearly dependent on the concentration of both TB13 and TRF. Monoclonal antibody TB13 did not inhibit the activities of B-cell stimulatory factor 1, interleukin 1, interleukin 2, or interleukin 3. TRF activity in dissolved samples of immunoprecipitates obtained with TB13 was recovered after NaDodSo4/PAGE, in the fractions corresponding to a protein band at Mr 46,000. Our results indicate that monoclonal antibody TB13 recognizes a molecule that has both TRF and BCGF-II activities.