Romani N, Inaba K, Puré E, Crowley M, Witmer-Pack M, Steinman R M
Laboratory of Cellular Physiology and Immunology, Rockfeller University, New York, New York 10021.
J Exp Med. 1989 Mar 1;169(3):1153-68. doi: 10.1084/jem.169.3.1153.
Resting T cells enter cell cycle when challenged with anti-CD3 mAb and accessory cells that bear required Fc receptors (FcR). Presentation of anti-CD3 is thought to be a model for antigens presented by accessory cells to the TCR complex. We have obtained evidence that the number of anti-CD3 molecules that are associated with the accessory cell can be very small. We first noticed that thymic dendritic cells and cultured, but not freshly isolated, epidermal Langerhans cells (LC) were active accessory cells for responses to anti-CD3 mAb. DNA synthesis was abrogated by a mAb to the FcR but not by mAb to other molecules used in clonally specific antigen recognition, i.e., class I and II MHC products or CD4 and CD8. The requisite FcR could be identified on the LC but in small numbers. Freshly isolated LC had 20,000 FcR per cell, while the more active cultured LC had only 2,000 sites, using 125I-anti-FcR mAb in quantitative binding studies. Individual LC had similar levels of FcR, as evidenced with a sensitive FACS. FcR could not be detected on T cells or within the dendritic cell cytoplasm, at the start of or during the mitogenesis response. When the response was assessed at 30 h with single cell assays, at least 20 T cells became lymphoblasts per added LC, and at least 8 T cells were synthesizing DNA while in contact with the LC in discrete cell clusters. To the extent that anti-CD3 represents a polyclonal model for antigen presentation to specific T cell clones, these results suggest two conclusions. First, only 200-300 molecules of ligand on dendritic cells may be required to trigger a T cell. Second, the maturation of LC in culture entails "sensitizing" functions other than ligand presentation (anti-CD3 on FcR) to clonotypic T cell receptors.
静息T细胞在受到抗CD3单克隆抗体和带有所需Fc受体(FcR)的辅助细胞刺激时进入细胞周期。抗CD3的呈递被认为是辅助细胞向TCR复合物呈递抗原的模型。我们已获得证据表明,与辅助细胞相关的抗CD3分子数量可能非常少。我们首先注意到,胸腺树突状细胞以及培养的而非新鲜分离的表皮朗格汉斯细胞(LC)是对抗CD3单克隆抗体反应的活跃辅助细胞。DNA合成被针对FcR的单克隆抗体阻断,但未被针对克隆特异性抗原识别中使用的其他分子的单克隆抗体阻断,即I类和II类MHC产物或CD4和CD8。在LC上可以鉴定出所需的FcR,但数量很少。在定量结合研究中,使用125I - 抗FcR单克隆抗体,新鲜分离的LC每个细胞有20,000个FcR,而活性更高的培养LC只有2,000个位点。单个LC具有相似水平的FcR,灵敏的荧光激活细胞分选术(FACS)证明了这一点。在有丝分裂反应开始时或期间,在T细胞或树突状细胞胞质内未检测到FcR。当在30小时用单细胞测定法评估反应时,每添加一个LC至少有20个T细胞变成淋巴母细胞,并且至少有8个T细胞在与离散细胞簇中的LC接触时正在合成DNA。就抗CD3代表向特定T细胞克隆呈递抗原的多克隆模型而言,这些结果表明两个结论。第一,树突状细胞上仅200 - 300个配体分子可能就足以触发T细胞。第二,培养的LC的成熟需要除了向克隆型T细胞受体呈递配体(FcR上的抗CD3)之外的“致敏”功能。