Kakkanaiah V N, Nagarkatti M, Nagarkatti P S
Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg 24061.
Cell Immunol. 1990 May;127(2):442-57. doi: 10.1016/0008-8749(90)90145-h.
Autoimmune-susceptible, MRL-lpr/lpr (lpr) mice develop a profound lymphadenopathy resulting from the accumulation of CD4-CD8- (double-negative, DN) cells in peripheral lymphoid organs. The source and the mechanism of this abnormal accumulation of cells is still unknown. Recently, we reported that a significant number (approximately 35%) of the CD4-CD8- cells expressed J11d, a marker expressed by immature thymocytes but not by mature functional peripheral T cells. In the present study, we investigated the phenotype, growth requirements, and functional properties of purified J11d+ and J11d- subpopulations. Using the mAb, F23.1, which recognizes a TCR determinant encoded by the V beta 8 gene family, it was observed that approximately 30% of the J11d+ and J11d- DN cells expressed this determinant. Further studies on the thymus revealed that J11d+ DN cells from lpr thymus also contained F23.1+ cells (approximately 25%), whereas, similar cells from normal MRL(-)+/+mice were all F23.1-, consistent with earlier reports in other normal strains. Further phenotypic studies revealed that the peripheral J11d+ and J11d- cells from lpr mice were similar in expressing CD3, Ly-5 (B220), and Ly-24 (Pgp-1) determinants. When stimulated with phorbol myristic acetate (PMA) and recombinant IL-2 (rIL-2), only J11d- cells but not J11d+ cells responded by proliferation. However, in the presence of calcium ionophore (A23187) and PMA, both J11d+ and J11d- subpopulations proliferated by producing and responding to endogenous IL-2 but not IL-4. The lymph node T cells from 1-month-old MRL-lpr/lpr mice responded strongly when stimulated with PMA + rIL-4 or PMA + rIL-6. In contrast both J11d+ and J11d- subpopulations failed to respond when similarly stimulated. The J11d+ but not J11d- cells demonstrated spontaneous cytotoxic activity against the NK-sensitive YAC-1 tumor targets. The J11d- cells did not exhibit cytotoxic potential in spite of culture with PMA + rIL-2. Even after repeated culture in vitro with PMA + A23187 or PMA + rIL-2, both J11d+ and J11d- subpopulations failed to express the mature phenotype bearing CD4 and/or CD8 antigens. The present study demonstrates the expansion of unique J11d+, alpha beta-TCR+, DN T cells with cytotoxic potential in lpr mice and further suggests the existence of phenotypic and functional heterogeneity among the abnormal lpr DN cells.
自身免疫易感的MRL-lpr/lpr(lpr)小鼠会因外周淋巴器官中CD4-CD8-(双阴性,DN)细胞的积累而出现严重的淋巴结病。这种细胞异常积累的来源和机制仍然未知。最近,我们报道了相当数量(约35%)的CD4-CD8-细胞表达J11d,这是一种由未成熟胸腺细胞表达但成熟功能性外周T细胞不表达的标志物。在本研究中,我们研究了纯化的J11d+和J11d-亚群的表型、生长需求和功能特性。使用识别由Vβ8基因家族编码的TCR决定簇的单克隆抗体F23.1,观察到约30%的J11d+和J11d- DN细胞表达了该决定簇。对胸腺的进一步研究表明,来自lpr胸腺的J11d+ DN细胞也含有F23.1+细胞(约25%),而来自正常MRL(-)+/+小鼠的类似细胞均为F23.1-,这与其他正常品系的早期报道一致。进一步的表型研究表明,来自lpr小鼠的外周J11d+和J11d-细胞在表达CD3、Ly-5(B220)和Ly-24(Pgp-1)决定簇方面相似。当用佛波醇肉豆蔻酸酯(PMA)和重组IL-2(rIL-2)刺激时,只有J11d-细胞而不是J11d+细胞通过增殖做出反应。然而,在存在钙离子载体(A23187)和PMA的情况下,J11d+和J11d-亚群均通过产生和响应内源性IL-2而非IL-4进行增殖。1月龄MRL-lpr/lpr小鼠的淋巴结T细胞在用PMA + rIL-4或PMA + rIL-6刺激时反应强烈。相比之下,当受到类似刺激时,J11d+和J11d-亚群均无反应。J11d+而非J11d-细胞对NK敏感的YAC-1肿瘤靶标表现出自发细胞毒性活性。尽管用PMA + rIL-2培养,J11d-细胞仍未表现出细胞毒性潜力。即使在用PMA + A23187或PMA + rIL-2进行体外反复培养后,J11d+和J11d-亚群均未能表达带有CD4和/或CD8抗原的成熟表型。本研究证明了lpr小鼠中具有细胞毒性潜力的独特J11d+αβ-TCR+ DN T细胞的扩增,并进一步表明异常的lpr DN细胞之间存在表型和功能异质性。