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用~3H-尿苷离体掺入实验研究兔腹腔巨噬细胞在识别抗原过程中的RNA合成。当用不同种类的红细胞膜作抗原时,无论是自体兔膜还是人、羊、异体兔的膜,都促进~3H-尿苷掺入巨噬细胞RNA,但程度上有所不同(异体兔>羊、人>自体兔)。然而,如果用利福平或放线菌素D预先完全抑制巨噬细胞正常转录之后,再用不同种类的红细胞膜刺激,结果发现自体膜不再能刺激巨噬细胞的RNA合成,而其他种类的膜都在一定程度上恢复巨噬细胞的RNA合成。用2.5％聚丙烯酰胺-0.5％琼脂糖凝胶电泳分析巨噬细胞在抗原刺激下新合成的RNA,结果表明上述几种红细胞膜都促进所有种类的RNA合成,但当用利福平完全抑制正常转录时,外来抗原只促进4—5 S RNA的合成。poly(U)-Sepharose亲和层析表明这种4—5 S RNA带有poly(A)。根据这些结果,提出了一个关于巨噬细胞识别抗原过程的工作假设。
In vitro incorporation of 3H-uridine was used to study the synthesis of RNA in rabbit peritoneal macrophages during antigen recognition. When different types of erythrocyte membranes were used as antigens, the incorporation of ~ 3H-uridine into macrophage RNA was promoted by either autologous rabbit membranes or membranes of human, sheep and allogeneic rabbits to varying degrees (allogeneic rabbits> Sheep, human> autologous rabbit). However, if rifampicin or actinomycin D were used to completely inhibit the normal transcription of macrophages before being stimulated with different types of erythrocyte membranes, it was found that autologous membranes were no longer able to stimulate macrophage RNA synthesis, whereas other types Of the membranes are to some extent restore macrophage RNA synthesis. The newly synthesized RNA of macrophages stimulated with antigen was analyzed by 2.5% polyacrylamide-0.5% agarose gel electrophoresis. The results showed that all the above-mentioned erythrocyte membranes promoted the synthesis of all kinds of RNAs, but when rifampin was completely inhibited During normal transcription, foreign antigens only promote the synthesis of 4-5 S RNA. Poly (U) -Sepharose affinity chromatography showed that this 4-5 S RNA carries poly (A). Based on these results, a working hypothesis on the process of antigen recognition by macrophages was proposed.