Taylor S G, Saffold P C, Braun D P, Harris J E
J Natl Cancer Inst. 1982 Apr;68(4):549-54.
Peripheral blood mononuclear cells (PBMC), T-cells, non-T-cells, TG cells, and TM cells were tested for leukocyte adherence inhibition (LAI) reactivity to purified protein derivative (PPD) in normal human volunteers to determine the reactive cell subfractions. T-cells were separated by sheep red blood cell rosetting and TM and TG cells by rosetting with anti-ox red blood cell (ORBC) IgM- and IgG-treated ORBC. Fresh PBMC, T-cells, and TG cells gave positive LAI reactivity to PPD in PPD-positive donors only. PBMC that had undergone 24-hour incubation or extensive washing, non-T-cells, and TM cells were unreactive. All cell subfractions were negative in PPD-negative donors. Indirect LAI assay was performed on the spent media from overnight incubation of unstimulated PBMC and also following a 1-hour incubation of the various cell subfractions obtained from a PPD-reactive donor with PPD. Positive reactions, indicating the release of an LAI factor (LAIF), were obtained with the spent media and with the T-cell and TG cell subfractions following PPD stimulation, whereas non-T-cell and TM cell subfractions yielded no reactivity. Incubation of sensitized T-cells with PPD in the presence of cycloheximide and preincubation of T-cells for 24 hours with cycloheximide before PPD exposure failed to inhibit LAI reactivity. These data suggest that T-cells, specifically TG cells, can release LAIF upon specific antigen stimulation with PPD, whereas non-T-cell and TM cell subpopulations fail to respond to the same stimulus. LAIF may also be released into the culture media with prolonged (overnight) incubation in artificial media or with extensive washing. Inhibition of protein synthesis during antigen exposure or up to 24 hours before antigen exposure does not appear to inhibit LAI reactivity.
在正常人类志愿者中,检测外周血单个核细胞(PBMC)、T细胞、非T细胞、TG细胞和TM细胞对纯化蛋白衍生物(PPD)的白细胞黏附抑制(LAI)反应性,以确定反应性细胞亚群。通过绵羊红细胞花环法分离T细胞,通过用抗氧红细胞(ORBC)IgM和IgG处理的ORBC花环法分离TM细胞和TG细胞。新鲜的PBMC、T细胞和TG细胞仅在PPD阳性供体中对PPD产生阳性LAI反应性。经过24小时孵育或大量洗涤的PBMC、非T细胞和TM细胞无反应性。所有细胞亚群在PPD阴性供体中均为阴性。对未刺激的PBMC过夜孵育后的用过的培养基以及对从PPD反应性供体获得的各种细胞亚群与PPD孵育1小时后进行间接LAI测定。PPD刺激后,用过的培养基以及T细胞和TG细胞亚群获得了阳性反应,表明释放了LAI因子(LAIF),而非T细胞和TM细胞亚群无反应性。在存在放线菌酮的情况下,将致敏T细胞与PPD一起孵育,以及在PPD暴露前用放线菌酮将T细胞预孵育24小时,均未能抑制LAI反应性。这些数据表明,T细胞,特别是TG细胞,在受到PPD特异性抗原刺激时可释放LAIF,而非T细胞和TM细胞亚群对相同刺激无反应。在人工培养基中长时间(过夜)孵育或大量洗涤后,LAIF也可能释放到培养基中。抗原暴露期间或抗原暴露前长达24小时的蛋白质合成抑制似乎并未抑制LAI反应性。