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Characterization of monoclonal nonspecific suppressor factor (MNSF) with the use of a monoclonal antibody.

使用单克隆抗体表征单克隆非特异性抑制因子 (MNSF)。

基本信息

批准号：
62480189
负责人：
TSUNEMATSU Tokugoro
金额：
$ 3.52万
依托单位：
Shimane Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62480189/
关键词：
Nonspecific Suppressor Factor 非特異抑性因子

项目摘要

In our previous study, we attempted to prepare a hybridoma producing a monoclonal nonspecific suppressor factor (MNSF). We attempted to define its biochemical characteristics, and then to compare it with already reported nonspecific suppressor factors. This MUSF inhibits the generation of lipopolysaccharide (LPS) induced immunoglobulin (Ig)-secreting cells. In the present study, we made a monoclonal antibody toward MNSF in murine system. The monoclonal antibody designated MO6 recognizes MNSF with specific reactivity. MO6 abrogates the suppressive effect of MNSF, whereas the murine IgG monoclonal antibodies against irrelevant antigens have no such effect. These results indicate that MO6 binds the functionally active site of MNSF and consequently block the biological activity. Immuno-blotting analysis showed that MO6 reacts with MNSF of 16,000 and 24,000 daltons, thereby, indicating that these species are identical, both functionally and immuno-logically. An explorative attempt was made to detect human nonspecific suppressor factor (HNSF) from the culture supernatant of Con A activated human peripheral blood mononuclear cells. The bound fraction obtained from the MO6 affinity column chromatography showed suppressor activity. This result indicates the existence of a suppressor factor in Con A activated supernatant, which is identical with murine MNSF in terms of biological activity. The purified HNSF shows charac-teristics similar to murine MNSF, that is, both have similar m.w. and physioc-chemical properties. We developed a ELISA system with the use of MO6 antibody in order to detect MNSF antigen. Furthere, N-terminal amino acid sequence of MNSF were analyzed. This information will be a pertinent probe for isolating the cDNA that encodes for MNSF.

在我们之前的研究中，我们试图制备一种产生单克隆性非特异性抑制因子(MNSF)的杂交瘤。我们试图定义它的生化特征，然后将其与已报道的非特异性抑制因子进行比较。该MUSF可抑制脂多糖(LPS)诱导的免疫球蛋白(Ig)分泌细胞的生成。在本研究中，我们在小鼠系统中制备了抗MNSF的单抗。命名为MO6的单抗识别MNSF具有特异的反应性。MO6可阻断MNSF的抑制作用，而抗无关抗原的小鼠免疫球蛋白单抗则无此作用。这些结果表明，MO6结合了MNSF的功能活性部位，从而阻断了其生物活性。免疫印迹分析表明，MO6与16,000和24,000道尔顿的MNSF反应，从而表明这两个物种在功能和免疫学上是相同的。对从ConA激活的人外周血单核细胞培养上清液中检测人非特异性抑制因子(HNSF)进行了探索性尝试。从MO6亲和柱层析得到的结合组分具有抑制活性。这一结果表明，ConA激活的上清液中存在抑制因子，其生物学活性与小鼠MNSF一致。纯化的HNSF显示出与小鼠MNSF相似的特征，即两者具有相似的分子量。和物理化学性质。为了检测MNSF抗原，我们利用MO6抗体建立了一种检测MNSF抗原的酶联免疫吸附试验系统。并对MNSF的N端氨基酸序列进行了分析。这一信息将为分离编码MNSF的cDNA提供一个有针对性的探针。