Peoples G E, Davey M P, Goedegebuure P S, Schoof D D, Eberlein T J
Department of Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
J Immunol. 1993 Nov 15;151(10):5472-80.
The interaction between T lymphocytes and the Ag-HLA complex on tumor cells is mediated by the TCR. The diversity and the specificity of the TCR are in part secondary to the gene rearrangement of the V region on the beta-chain (V beta). To determine whether a restricted number of TCR V beta genes are utilized in the recognition of ovarian cancer, tumor-infiltrating lymphocytes (TIL) were isolated from six consecutive untreated ovarian cancer patients. TIL were also cultured using repeated autologous tumor stimulation, and by 7 wk, five of six patients produced bulk cultures consisting of > 50% CD8+ T cells and demonstrating an autologous tumor-specific pattern of cytotoxicity. TCR V beta gene usage was analyzed in the five patients yielding fresh TIL and corresponding 7-wk cultured, tumor-specific TIL; 22 primers specific for 20 TCR V beta gene families were employed and amplified by polymerase chain reaction and then quantitated by HPLC. A heterogeneous pattern of V beta usage was seen in the fresh TIL; however, V beta 2, V beta 3, V beta 6, V beta 7, V beta 8, and V beta 13.1 were found in increased proportions in at least three of five patients. In the 7-wk tumor-specific TIL, V beta analysis showed an increased usage of V beta 2, V beta 3, V beta 6, and V beta 7 in more than three of five patients. No significant change in V beta representation was seen in control populations that were not stimulated with tumor. Looking at the percent change in V beta usage between fresh and 7-wk tumor-specific cultures, V beta 2 and V beta 6 were augmented significantly in at least three of five patients (108% and 61%, respectively). To verify that the increase in representation of these V beta families was responsible for the increased cytotoxicity observed, mAb specific for V beta 2 and V beta 6 were used to block tumor lysis. Anti-V beta 6 and anti-V beta 2 significantly blocked cytotoxicity against autologous tumor cells in those TIL populations expressing increased levels of these V beta families. These data suggest that a selective repertoire of TCR V beta genes is used to recognize the Ag-HLA class 1 complexes on the surface of ovarian tumor cells, and specifically V beta 2 and V beta 6 appear to mediate antitumor activity. These findings may aid in the development of a more specific immunotherapy in ovarian cancer.
T淋巴细胞与肿瘤细胞上的抗原 - 人类白细胞抗原（Ag - HLA）复合物之间的相互作用是由T细胞受体（TCR）介导的。TCR的多样性和特异性部分归因于β链（Vβ）上V区的基因重排。为了确定在卵巢癌的识别中是否使用了有限数量的TCR Vβ基因，从6例未经治疗的连续卵巢癌患者中分离出肿瘤浸润淋巴细胞（TIL）。TIL也通过反复的自体肿瘤刺激进行培养，到7周时，6例患者中有5例产生了大量培养物，其中CD8 + T细胞占比超过50%，并表现出针对自体肿瘤的特异性细胞毒性模式。对5例产生新鲜TIL以及相应的培养7周的肿瘤特异性TIL的患者进行了TCR Vβ基因使用情况分析；使用了针对20个TCR Vβ基因家族的22种引物，通过聚合酶链反应进行扩增，然后通过高效液相色谱进行定量分析。在新鲜TIL中观察到Vβ使用的异质性模式；然而，在5例患者中的至少3例中发现Vβ2、Vβ3、Vβ6、Vβ7、Vβ8和Vβ13.1的比例增加。在培养7周的肿瘤特异性TIL中，Vβ分析显示在5例患者中的3例以上，Vβ2、Vβ3、Vβ6和Vβ7的使用增加。在未用肿瘤刺激的对照群体中，Vβ表达没有显著变化。观察新鲜和培养7周的肿瘤特异性培养物之间Vβ使用的百分比变化，在5例患者中的至少3例中，Vβ2和Vβ6显著增加（分别为108%和61%）。为了验证这些Vβ家族表达的增加是否是观察到的细胞毒性增加的原因，使用针对Vβ2和Vβ6的单克隆抗体来阻断肿瘤溶解。在那些表达这些Vβ家族水平增加的TIL群体中，抗Vβ6和抗Vβ2显著阻断了针对自体肿瘤细胞的细胞毒性。这些数据表明，TCR Vβ基因的选择性库用于识别卵巢肿瘤细胞表面的Ag - HLA Ⅰ类复合物，特别是Vβ2和Vβ6似乎介导了抗肿瘤活性。这些发现可能有助于开发更具特异性的卵巢癌免疫疗法。
