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

采用抗杂交瘤源性非特异性抑制因子（MNSF）单克隆抗体对其进行纯化。在还原条件下对纯化的天然MNSF进行sds -聚丙烯酰胺凝胶电泳，然后进行免疫印迹转移和氨基黑化学染色。切除70 kda条带，进行氨基酸序列分析。测定了18个氨基酸的n端序列。合成了5个氨基酸对应的退化引物，用于RF-PCR。MNSFalpha mRNA在cona激活（12小时）的T细胞中表达，而在未刺激的T细胞中不表达。从部分MNSFalpha cDNA中推断出的氨基酸序列与任何已知的蛋白质都没有同源性。然而，该序列与T细胞受体(TCR) α链有35%的同源性，这与MNSFalpha可能与TCR α链相关的假设很好地吻合。MNSFalpha全cDNA测序正在进行中。在不久的将来，利用重组MNSFalpha来阐明MNSF的作用模式应该是可能的。

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.