Guha A, Cone R E
Department of Pathology (Division of Experimental Pathology), University of Connecticut Health Center, Farmington 06032.
J Immunol. 1991 Mar 15;146(6):1858-61.
The effect of IFN-gamma on the rate of shedding and biosynthesis of H-2Dd was determined by culture of cell surface-radioiodinated BALB/c spleen cells with rIFN-gamma or spleen cells metabolically labeled with 35S-methionine in the presence of IFN-gamma. Radioiodinated or 35S-labeled H-2Dd was quantitated by immunoprecipitation of H-2Dd from detergent lysates of radiolabeled cells taken at different culture intervals. The loss of 125I-labeled H-2Dd was retarded 75 to 90% by IFN-gamma whereas the biosynthetic rate was unaffected during the first 10-h culture. The net result was a ninefold increase in newly synthesized cell-associated H-2Dd. The results were consistent with determination of the kinetics of increased expression of H-2Dd determined by immunofluorescence and suggest that an early effect of IFN-gamma on the expression of class I Ag is a retardation of catabolism leading to an increase of newly synthesized class I Ag.
通过用重组干扰素-γ(rIFN-γ)培养细胞表面放射性碘化的BALB/c脾细胞,或在干扰素-γ存在的情况下用35S-甲硫氨酸代谢标记脾细胞,来测定干扰素-γ对H-2Dd脱落率和生物合成的影响。通过对在不同培养间隔时取自放射性标记细胞的去污剂裂解物中的H-2Dd进行免疫沉淀,对放射性碘化或35S标记的H-2Dd进行定量。干扰素-γ使125I标记的H-2Dd的丢失延迟了75%至90%,而在最初10小时的培养过程中生物合成速率未受影响。最终结果是新合成的细胞相关H-2Dd增加了九倍。这些结果与通过免疫荧光测定的H-2Dd表达增加的动力学结果一致,并表明干扰素-γ对I类抗原表达的早期作用是减缓分解代谢,导致新合成的I类抗原增加。