Recognition of HLA-A2 mutant and variant target cells by an HLA-A2 allospecific human cytotoxic T lymphocyte line.
Recognition of HLA-A2 mutant and variant target cells by an HLA-A2 allospecific human cytotoxic T lymphocyte line.
复制标题
HLA-A2 同种异体特异性人细胞毒性 T 淋巴细胞系对 HLA-A2 突变体和变异靶细胞的识别。
作者：
C. Ware;M. Krangel;D. Pious;S. Burakoff;J. Strominger
用T细胞生长因子和外周血淋巴细胞与选定的同种异体靶细胞共培养，建立了人类白细胞抗原A2特异性细胞毒性T淋巴细胞(CTL)细胞系。CTL-8细胞系在培养5周后与14个淋巴母细胞系进行51铬(51Cr)释放试验，显示出对携带人类白细胞抗原(HL A)-A2的靶细胞的特异性。纯化的抗人白细胞抗原(HLA)单抗W6/32和PA2.1对细胞溶解的抑制率分别为85%和60%。CTL-8细胞系在凝集素刀豆蛋白A(ConA)或植物血凝素-P凝集素(PHA-P)存在的情况下裂解非人类白细胞抗原-A2靶细胞，表明细胞溶解的特异性不是由于靶细胞对CTL裂解机制的非特异性抵抗所致。以T5-1人类白细胞抗原A2突变细胞系作为CTL-8细胞系的靶细胞。细胞克隆8.18.1、8.21.1和8.6.1与T5-1野生型一样有效地裂解细胞克隆8.18.1、8.21.1和8.6.1，这些克隆表达改变的人类白细胞抗原A2分子，这是因为它们与同种特异性单抗的反应性降低。这些细胞系也是正常的人类白细胞抗原A2靶细胞的有效冷靶竞争者。在直接细胞溶解和冷靶竞争抑制实验中，8.14.1细胞克隆表达较少的人类白细胞抗原A2同种异体抗原，与CTL-8的反应性相应降低。相反，表达正常的HLA-A2血清学决定簇的M7和DK1突变细胞株不能被CTL-8有效地裂解，并且不能作为正常的HLA-A2靶细胞的竞争性抑制物。这些结果支持T细胞和抗体识别的同种异体抗原决定簇(S)存在于人类白细胞抗原A2分子上的不同区域的观点。
HLA-A2 specific human cytotoxic T lymphocytes (CTL) cell lines have been developed using T cell growth factor and coculture of peripheral blood lymphocytes with selected allogeneic target cell lines. The CTL-8 line showed specificity for human leukocyte antigens (HLA)-A2 bearing target cells after 5 weeks in culture when tested against a panel of 14 lymphoblastoid cell lines in a 51Chromium (51Cr) release assay. Purified anti-human leukocyte antigens (HLA) monoclonal antibodies W6/32 and PA2.1 inhibited cytolysis by 85% and 60%, respectively. The CTL-8 line lysed non-HLA-A2 target cells in the presence of lectins concanavalin A (Con A) or phytohemagglutinin-P lectin (PHA-P) indicating the specificity of cytolysis was not due to nonspecific resistance of target cells to the CTL-lytic mechanism. The T5-1 HLA-A2 mutant cell series were tested as targets for the CTL-8 line. Cell clones 8.18.1, 8.21.1 and 8.6.1, which express altered HLA-A2 molecules as determined by their decreased reactivity with allospecific monoclonal antibodies, were lysed by the CTL-8 line as efficiently as the T5-1 wild type. These cell lines also acted as efficient cold target competitors for a normal HLA-A2 target cell. The 8.14.1 cell clone expressed a lower amount of HLA-A2 alloantigen and showed a corresponding decreased reactivity with CTL-8 in direct cytolytic and cold target competitive inhibition assays. In contrast, the M7 and DK1 HLA-A2 variant cell lines, which express normal HLA-A2 serological determinants, were inefficiently lysed by CTL-8 and did not act as competitive inhibitors of normal HLA-A2 target cells. These results support the concept that the alloantigenic determinant(s) recognized by T cells and antibodies occur at separate regions on the HLA-A2 molecule.