CORRELATION BETWEEN CD8 DEPENDENCY AND DETERMINANT DENSITY USING PEPTIDE-INDUCED, LD-RESTRICTED CYTOTOXIC LYMPHOCYTES-T
CORRELATION BETWEEN CD8 DEPENDENCY AND DETERMINANT DENSITY USING PEPTIDE-INDUCED, LD-RESTRICTED CYTOTOXIC LYMPHOCYTES-T
复制标题
DOI：
10.1084/jem.173.4.849
复制
发表时间：
1991-04-01
影响因子：
15.3
通讯作者：
CONNOLLY, JM
中科院分区：
文献类型：
--
作者：
ALEXANDER, MA;DAMICO, CA;CONNOLLY, JM
我们利用H-2L(D)的一些独特性质来研究细胞毒性T淋巴细胞(CTL)启动与效应功能的决定簇密度要求，并将决定簇密度要求与CD8依赖性相关联。在先前的研究中(Lie，W.-R.，N.B.Myers，J.Gorka，R.J.Rubocki，J.M.Connolly和T.H.Hansen)。1990年。大自然[长]。344：439)，我们证明了用已知受H-2L(D)限制的多肽培养正常细胞后，表面L(D)的表达增加了两到四倍。在本研究中，我们通过用小鼠巨细胞病毒或肿瘤多肽培养脾细胞来产生L(D)限制性的、多肽特异性的体外原代CTL，该浓度可以最大限度地诱导L(D)的表达。这些CTL可以用比初级致敏所需的更低剂量的多肽来敏化靶细胞以便识别。这表明启动和效应器功能的决定因素密度要求不同。利用多肽产生的体外原代CTL对内源性表达决定簇的靶细胞具有较弱的杀伤作用，很容易获得具有较强内源性表达的CTL株和克隆。在这两种情况下，用外源性多肽处理的靶细胞比内源性表达抗原的靶细胞裂解得更好。这表明，在多肽喂养和内源性靶点的决定簇密度上存在差异。当观察到表达内源性决定簇的靶细胞的裂解水平与不同TOM特异性CTL克隆识别靶细胞所需的多肽量成反比时，这种解释就得到了证实。此外，用于治疗靶细胞的多肽和CD8抗体的同时滴定显示，所分析的各种CTL克隆显示出非常不同的CD8依赖性。在每种情况下，CD8依赖与决定簇密度需求呈负相关。因此，CTL对CD8的依赖是相对的，但与其对决定簇密度的依赖表现出绝对的定量相关性。这些发现表明，在只可能遇到低决定簇密度的生理条件下，所有CTL克隆都将至少表现出部分CD8依赖。
We have taken advantage of some unique properties of H-2L(d) to investigate the determinant density requirements for cytotoxic T lymphocyte (CTL) priming versus effector function and to correlate the determinant density requirements with CD8 dependency. In a previous study (Lie, W.-R., N. B. Myers, J. Gorka, R. J. Rubocki, J. M. Connolly, and T. H. Hansen. 1990. Nature [Lond.]. 344:439), we demonstrated that culturing normal cells with peptides known to be restricted by H-2L(d) led to a two- to fourfold increase in surface L(d) expression. In the present study, we demonstrate the generation of L(d)-restricted, peptide-specific in vitro primary CTL by culturing spleen cells with murine cytomegalovirus or tum- peptide at concentrations previously shown to result in maximum induction of L(d) expression. Target cells can be sensitized for recognition by these CTL with lower doses of peptide than are required for the primary sensitization. This demonstrates differences in the determinant density requirements for priming versus effector function. The in vitro primary CTL generated with peptide can weakly lyse target cells that express the determinant endogenously, and CTL lines and clones capable of strong lysis of endogenous expressors are easily obtained. In both cases, target cells treated with exogenous peptide are lysed better than target cells expressing antigen endogenously. This suggested that there are differences in the determinant density of peptide-fed versus endogenous targets. This interpretation was substantiated when it was observed that the level of lysis of target cells expressing endogenous determinants correlated inversely with the amount of peptide required to sensitize targets for recognition by various tum- -specific CTL clones. Furthermore, simultaneous titration of both the peptide used to treat target cells and the antibody to CD8 revealed that the various CTL clones analyzed displayed widely disparate CD8 dependencies. In each case, the CD8 dependency correlated inversely with the determinant density requirement. Therefore, CD8 dependency of CTL is relative, but shows an absolute and quantitative correlation with their dependency on determinant density. These findings suggest that under physiologic conditions, where only low determinant densities are likely to be encountered, all CTL clones will show at least partial CD8 dependency.