STUDY ON THE MODE OF ACTION OF MONOCLONAL NONSPECIFIC SUPPRESSOR FACTOR (MNSF)

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Hybridoma-derived monoclonal nonspecific suppressor factor (MNSF) was purified with the use of anti-MNSF monoclonal antibody. SDS-polyacryl amide gel electrophoresis of the purified native MNSF was performed under reducing conditions, followed by immunoblot transfer and chemical staining with Amino black. The 70-kDa band was excised, and subjected to amino acid sequence analysis. The N-terminal sequence of 18 amino acids was determined. Degenerated primers corresponding to five amino acids were synthesized and used for RF-PCR.MNSFalpha mRNA was expressed in concanavalin A (Con A) activated (12 hours ) -, but not in unstimulated- T cells. The amino acid sequences deduced from the partial MNSFalpha cDNA showed no homology to any known proteins. However, the sequence showed 35% homology to T cell receptor (TCR) alpha chain in good aggrement with a hypothesis that MNSFalpha might be related to TCRalpha chain. The sequencing of full MNSFalpha cDNA was under way. The elucidation of the mode of action of MNSF with the use of recombinant MNSFalpha should be possible, in the near future.