Analysis of the functional capabilities of CD3+CD4-CD8- and CD3+CD4+CD8+ human T cell clones.
Analysis of the functional capabilities of CD3+CD4-CD8- and CD3+CD4+CD8+ human T cell clones.
复制标题
DOI：
10.4049/jimmunol.143.4.1108
复制
发表时间：
1989-08
影响因子：
4.4
通讯作者：
S. Patel;M. Wacholtz;A. Duby;D. Thiele;P. Lipsky
中科院分区：
文献类型：
--
作者：
S. Patel;M. Wacholtz;A. Duby;D. Thiele;P. Lipsky
检测人外周血CD3+CD4-CD8-和CD3+CD4+CD8+ T细胞克隆的功能。在PHA、rIL-2存在的条件下，用纯化的CD3+CD4-CD8-和CD3+CD4+CD8+ T细胞在极限稀释（0.3个细胞/孔）下培养，并将PBMC作为饲料照射，产生克隆。共生成12个CD3+CD4-CD8-克隆和5个CD3+CD4+CD8+克隆。通过分析TCR γ链和β链重排模式证明了克隆性。所有的CD3+CD4-CD8-克隆都用鉴定TCR δ链框架表位的TCR- δ 1单抗染色，但用鉴定成熟T细胞上TCR- α - β的WT31单抗染色无效。相比之下，CD3+CD4+CD8+克隆均被WT31染色，而不被TCR-delta 1染色。对17个无性系进行了各种功能活性筛选。当用固定的单抗刺激CD3时，它们分泌IL-2、ifn - γ和淋巴素/ tnf样因子（64.1），尽管数量不同。这些克隆分泌的IL-2和ifn - γ远低于表达CD3+CD4+CD8-或CD3+CD4-CD8 -CD8+ α β的克隆，但淋巴毒素/肿瘤坏死因子的含量相当。所有克隆也都具有不受mhc限制的细胞毒细胞的功能。这种活性与CD3+CD4+CD8-或CD3+CD4-CD8+ α - β克隆介导的活性相当。12个CD3+CD4-CD8-克隆中的9个和5个CD3+CD4+CD8+克隆中的4个在被固定的抗CD3激活时能够支持B细胞分化，但通常不如CD3+CD4+CD8-或CD3+CD4-CD8+ α β克隆有效。不同克隆的功能能力的差异不能用CD3分子复合物信号传导能力的改变来解释，因为mAb对CD3诱导的细胞内游离钙在每个被检查的克隆中都有相当的增加。当用PWM刺激克隆时，每个克隆都抑制了丝裂霉素c处理的新鲜CD4+ T淋巴细胞支持的B细胞分化。抑制依赖于添加到培养中的克隆细胞的数量，但可以观察到每微滴孔中只有12,500个细胞。表型分析显示，所有克隆均表达CD29、CD11b和NKH1表面Ag。这些结果表明，CD3+CD4-CD8-和CD3+CD4+CD8+ T细胞克隆表现出成熟T细胞的许多功能特征，尽管它们产生IL-2和ifn - γ，并对B细胞分化的帮助不如CD3+CD4+CD8-和CD3+CD4-CD8+ α β T细胞克隆有效。
The functional capabilities of human peripheral blood CD3+CD4-CD8- and CD3+CD4+CD8+ T cell clones were examined. The clones were generated by culturing purified populations of CD3+CD4-CD8- and CD3+CD4+CD8+ T cells at limiting dilution (0.3 cell/well) in the presence of PHA, rIL-2, and irradiated PBMC as feeders. Twelve CD3+CD4-CD8- and 5 CD3+CD4+CD8+ clones were generated. Clonality was documented by analyzing TCR gamma- and beta-chain rearrangement patterns. All CD3+CD4-CD8- clones were stained by the TCR-delta 1 mAb that identifies a framework epitope of the TCR delta-chain, but not by mAb WT31 that identifies the TCR-alpha beta on mature T cells. In contrast, the CD3+CD4+CD8+ clones were all stained by WT31 and not by TCR-delta 1. All 17 clones were screened for various functional activities. Each secreted IL-2, IFN-gamma, and lymphotoxin/TNF-like factors when stimulated with immobilized mAb to CD3 (64.1), albeit in varying quantities. These clones secreted far less IL-2 and IFN-gamma than CD3+CD4+CD8- or CD3+CD4-CD8+ alpha beta expressing clones, but comparable amounts of lymphotoxin/TNF. All clones also functioned as MHC-unrestricted cytotoxic cells. This activity was comparable to that mediated by the CD3+CD4+CD8- or CD3+CD4-CD8+ alpha beta clones. Nine of 12 CD3+CD4-CD8- and 4 of 5 CD3+CD4+CD8+ clones were able to support B cell differentiation when activated by immobilized anti-CD3, but usually not as effectively as the CD3+CD4+CD8- or CD3+CD4-CD8+ alpha beta clones. The differences in the functional capabilities of the various clones could not be accounted for by alterations in the signaling capacity of the CD3 molecular complex as mAb to CD3 induced comparable increases in intracellular free calcium in each clone examined. When clones were stimulated with PWM, each suppressed B cell differentiation supported by mitomycin C-treated fresh CD4+ T lymphocytes. Suppression was dependent on the number of clone cells added to culture, but could be observed with as few as 12,500 cells per microtiter well. Phenotypic analysis of the clones revealed that all expressed CD29, CD11b, and the NKH1 surface Ag. These results demonstrate that the CD3+CD4-CD8- and CD3+CD4+CD8+ T cell clones exhibit many of the functional characteristics of mature T cells, although they produce IL-2 and IFN-gamma and provide help for B cell differentiation less effectively than CD3+CD4+CD8- and CD3+CD4-CD8+ alpha beta T cell clones.