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戊二醛固定靶细胞的合成能力及抗原表达研究

Studies on the synthetic capacity and antigenic expression of glutaraldehyde-fixed target cells.

作者信息

Bubbers J E, Henney C S

出版信息

J Immunol. 1975 Mar;114(3):1126-31.

PMID:122990
Abstract

Mastocytoma cells (P815 of the DBA/2 strain) treated with increasing concentrations of glutaraldehyde were concurrently evaluated for their ability to incorporate exogeneous uridine, thymidine, and amino acids. The antigenic expression and membrane integrity of these treated cells were assayed by measuring their susceptibility to lysis by antibody and complement and by T-effector cells. The concentrations of glutarladehyde required to effect target cell antigen display were greater than those required to inhibit totally the cell's protein and nucleic acid synthetic processes. Thus, P815 cells treated with 0.15% glutaraldehyde for 10 sec were unable to incorporate either amino acids or nucleotides into macromolecules, but were readily lysed by effector T cell populations, and by alloantibody in the presence of complement. Target cells treated with glutaraldehyde concentrations in excess of 0.25% for 10 sec were resistant to both forms of immune lysis. In keeping with these observations, monolayers of P815 cells treated for 10 sec with 0.15% glutaraldehyde, were capable of specifically depleting T-effector cells from a cytolytically-active spleen cell population. After treatment with higher concentrations of glutaraldehyde (0.3%), however, the monolayers lost their capacity to adsorb effector cells. Although P815 cells treated with glutaraldehyde continued to exhibit H-2d alloantigen, neither these cells nor glutaraldehyde-treated DBA/2 spleen cells induced significant blastogenesis or stimulated the production of cytolytically active effector cells in mixed leukocyte cultures.

摘要

用浓度不断增加的戊二醛处理肥大细胞瘤细胞(DBA/2品系的P815细胞),同时评估其摄取外源性尿苷、胸苷和氨基酸的能力。通过测量这些处理过的细胞对抗体、补体和T效应细胞裂解的敏感性,来检测其抗原表达和膜完整性。实现靶细胞抗原展示所需的戊二醛浓度高于完全抑制细胞蛋白质和核酸合成过程所需的浓度。因此,用0.15%戊二醛处理10秒的P815细胞无法将氨基酸或核苷酸掺入大分子中,但很容易被效应T细胞群体以及在补体存在下的同种异体抗体裂解。用超过0.25%的戊二醛浓度处理10秒的靶细胞对两种形式的免疫裂解均具有抗性。与这些观察结果一致,用0.15%戊二醛处理10秒的P815细胞单层能够从具有细胞溶解活性的脾细胞群体中特异性耗尽T效应细胞。然而,用更高浓度的戊二醛(0.3%)处理后,单层细胞失去了吸附效应细胞的能力。尽管用戊二醛处理的P815细胞继续表现出H-2d同种异体抗原,但在混合白细胞培养中,这些细胞以及用戊二醛处理的DBA/2脾细胞均未诱导明显的母细胞化或刺激产生具有细胞溶解活性的效应细胞。

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