| Budget Amount *help |
¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1988: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1987: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Research Abstract |
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.
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