Peoples G E, Goedegebuure P S, Andrews J V, 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):5481-91.
Tumor-associated lymphocytes (TAL) from the malignant ascites and tumor-infiltrating lymphocytes (TIL) from the solid tumor were isolated from six consecutive untreated ovarian cancer patients. Tumor-specific CTL were generated from both TAL and TIL using solid phase anti-CD3, low dose IL-2 (50 IU/ml), and repeated tumor stimulation. The specificity of TAL and TIL was tested in standard cytotoxicity assays using autologous tumor, several allogeneic ovarian tumors, and the NK-sensitive cell line, K562. Anti-HLA-A-B-C mAb, W6/32, was used to demonstrate that these tumor-specific TAL and TIL were HLA class I-restricted. The ability of the ascitic and solid tumor to present Ag by HLA class I was assessed using Brefeldin A, a fungal metabolite that blocks the endogenous Ag-processing pathway in the viral model. Brefeldin A significantly inhibited tumor-specific cytotoxicity as well as HLA class I expression on the cell surface, suggesting an endogenous source of tumor-associated Ag. Despite previous reports of antigenic heterogeneity in ovarian cancer, shared tumor-associated Ag were shown to exist in this disease as demonstrated by significant allogeneic recognition of HLA-A2-matched patients as opposed to unmatched controls. Specifically, CTL from HLA-A2+ patients lysed HLA-A2+ allogeneic targets significantly better than HLA-A2- allogeneic or HLA-A2+ melanoma targets. There was no such difference with HLA-A2- effectors. Furthermore, HLA-A2 was confirmed to be a major restriction element in ovarian cancer by the blocking of HLA-A2+ effectors against both autologous and allogeneic HLA-A2+ targets with the anti-HLA-A2 mAb, BB7.2. These findings verify a similar lymphocyte/tumor interaction as has been documented in melanoma, suggesting a common mechanism of recognition of these human tumors by lymphocytes.
从6例未经治疗的连续卵巢癌患者中分离出恶性腹水中的肿瘤相关淋巴细胞（TAL）和实体瘤中的肿瘤浸润淋巴细胞（TIL）。使用固相抗CD3、低剂量白细胞介素-2（50 IU/ml）和重复的肿瘤刺激，从TAL和TIL中产生肿瘤特异性细胞毒性T淋巴细胞（CTL）。使用自体肿瘤、几种同种异体卵巢肿瘤以及NK敏感细胞系K562，在标准细胞毒性试验中测试TAL和TIL的特异性。使用抗HLA-A-B-C单克隆抗体W6/32来证明这些肿瘤特异性TAL和TIL受HLA I类分子限制。使用布雷菲德菌素A评估腹水和实体瘤通过HLA I类分子呈递抗原的能力，布雷菲德菌素A是一种真菌代谢产物，可阻断病毒模型中的内源性抗原加工途径。布雷菲德菌素A显著抑制肿瘤特异性细胞毒性以及细胞表面HLA I类分子的表达，提示肿瘤相关抗原的内源性来源。尽管先前有关于卵巢癌抗原异质性的报道，但与不匹配的对照相比，HLA-A2匹配患者之间显著的同种异体识别表明，该疾病中存在共享的肿瘤相关抗原。具体而言，来自HLA-A2+患者的CTL对HLA-A2+同种异体靶标的杀伤作用明显优于HLA-A2-同种异体或HLA-A2+黑色素瘤靶标。HLA-A2-效应细胞则无此差异。此外，通过用抗HLA-A2单克隆抗体BB7.2阻断HLA-A2+效应细胞对自体和同种异体HLA-A2+靶标的作用，证实HLA-A2是卵巢癌中的主要限制元件。这些发现证实了与黑色素瘤中所记录的类似的淋巴细胞/肿瘤相互作用，提示淋巴细胞识别这些人类肿瘤的共同机制。
