Inamitsu T
Department of Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka.
Fukuoka Igaku Zasshi. 1991 Feb;82(2):59-70.
HLA-DQw6 transgenic C57BL/6 mice (DQw6-B6) were utilized to define the role of HLA-DQw6 gene product in immune-recognition. To investigate the responsiveness of lymph node cells from C57BL/6 mice (B6) or DQw6-B6 in response to DQw6 molecules expressed in the DQw6-B6, in vitro secondary MLR was performed. The lymph node cells from B6 proliferated in response to spleen cells from DQw6-B6, whereas those from DQw6-B6 did not. These results suggested that DQw6-B6 acquired tolerance to DQw6 molecules. The difference of T cell repertoire between B6 and DQw6-B6 was investigated using mAbs directed against T cell receptor V beta regions, V beta 3, V beta 5, V beta 6, V beta 8, V beta 11 and V alpha 3.2. Although the proportion of V beta 5+ CD8+ T cells and V beta 6+ CD4+ T cells were increased in the B6 anti-DQw6-B6 MLR T cell line, there was no significant difference in the proportion of peripheral T cells expressing each V alpha or V beta region between B6 and DQw6-B6. Both CD4+ and CD8+ long term-cultured T lymphocyte cell lines were generated from lymph node cells of B6 stimulated in vitro by irradiated spleen cells from DQw6-B6. The CD4+ T cell line proliferated in response to spleen cells of DQw6-B6 or L cell transfectant expressing HLA-DQw6 molecules and these responses were completely inhibited by either anti-DQ or anti-CD4 monoclonal antibodies (mAbs) but not by anti I-Ab mAbs. The CD8+ T cell line lysed splenic cells activated by lipopolysaccharide (LPS) from DQw6-B6 spleen cells but not from B6. The CD8+ T cell line also exhibited a cytotoxicity to splenic LPS blast cells from backcross progenies between (DQw6-B6 x DBA1) F1 and DBA1, only when target cells expressed both HLA-DQw6 molecules and H-2b. These observations indicated that recognition of HLA-DQw6 by the CD8+ cytotoxic T cell line was restricted by H-2b. The cytolytic activity of the CD8+ T cell line was inhibited by either anti-CD8 or anti-H-2Db mAbs but not by anti-HLA-DQ nor anti-H-2Kb mAbs. These results show that HLA-DQ transgene products expressed in DQw6-B6 can induce xenogeneic MLR in both CD4+ and CD8+ T cells. The CD4+ T lymphocytes recognize the HLA-DQw6 molecule itself whereas the CD8+ cytotoxic T lymphocytes recognize the HLA-DQw6 gene product in the context of H-2Db.
利用 HLA - DQw6 转基因 C57BL/6 小鼠（DQw6 - B6）来确定 HLA - DQw6 基因产物在免疫识别中的作用。为了研究 C57BL/6 小鼠（B6）或 DQw6 - B6 的淋巴结细胞对 DQw6 - B6 中表达的 DQw6 分子的反应性，进行了体外二次混合淋巴细胞反应（MLR）。B6 的淋巴结细胞对 DQw6 - B6 的脾细胞有增殖反应，而 DQw6 - B6 的淋巴结细胞则无此反应。这些结果表明 DQw6 - B6 对 DQw6 分子获得了耐受性。使用针对 T 细胞受体 Vβ区域、Vβ3、Vβ5、Vβ6、Vβ8、Vβ11 和 Vα3.2 的单克隆抗体（mAb）研究了 B6 和 DQw6 - B6 之间 T 细胞库的差异。尽管在 B6 抗 DQw6 - B6 MLR T 细胞系中 Vβ5 + CD8 + T 细胞和 Vβ6 + CD4 + T 细胞的比例增加，但 B6 和 DQw6 - B6 之间表达各 Vα或 Vβ区域的外周 T 细胞比例没有显著差异。通过用 DQw6 - B6 的经照射脾细胞体外刺激 B6 的淋巴结细胞，产生了 CD4 + 和 CD8 + 长期培养的 T 淋巴细胞系。CD4 + T 细胞系对 DQw6 - B6 的脾细胞或表达 HLA - DQw6 分子的 L 细胞转染子有增殖反应，并且这些反应被抗 DQ 或抗 CD4 单克隆抗体完全抑制，但不被抗 I - Ab 单克隆抗体抑制。CD8 + T 细胞系裂解由 DQw6 - B6 脾细胞的脂多糖（LPS）激活的脾细胞，但不裂解 B6 的脾细胞。CD8 + T 细胞系对（DQw6 - B6×DBA1）F1 和 DBA1 之间回交后代的脾 LPS 母细胞也表现出细胞毒性，仅当靶细胞同时表达 HLA - DQw6 分子和 H - 2b 时。这些观察结果表明 CD8 + 细胞毒性 T 细胞系对 HLA - DQw6 的识别受 H - 2b 限制。CD8 + T 细胞系的细胞溶解活性被抗 CD8 或抗 H - 2Db 单克隆抗体抑制，但不被抗 HLA - DQ 或抗 H - 2Kb 单克隆抗体抑制。这些结果表明在 DQw6 - B6 中表达的 HLA - DQ 转基因产物可在 CD4 + 和 CD8 + T 细胞中诱导异种 MLR。CD4 + T 淋巴细胞识别 HLA - DQw6 分子本身，而 CD8 + 细胞毒性 T 淋巴细胞在 H - 2Db 的背景下识别 HLA - DQw6 基因产物。
