Tong A W, Lee J, Stone M J
Cancer Res. 1984 Nov;44(11):4987-92.
Two human small cell lung carcinoma cell lines, NCI-H69 and NCI-H128, were used as alternating sources of immunogen to generate monoclonal antibodies to small cell lung carcinoma-associated antigens. BALB/c mice were sensitized with seven injections of live tumor cells, four with NCI-H69 cells and three with NCI-H128 cells. Somatic cell hybridization was performed by fusion of the immune murine splenocytes using syngeneic myeloma cells from the SP2/0 Ag14 cell line. Hybridoma colonies were screened against small cell lung carcinoma cells and normal lung fibroblasts with an enzyme-linked immunosorbent assay. Compared to animals immunized with only NCI-H69 or NCI-H128 cells, alternate immunization resulted in the generation of a significantly higher number of hybridomas that reacted selectively with both tumor cell lines. Monoclonal antibodies from two reactive hybrid clones generated by alternate immunization, SCLC 2051 and SCLC 5023, were uniformly negative to normal human tissues including lung, kidney, liver, spleen, breast, thyroid, brain, small intestine, and colon. While both monoclonal antibodies were nonreactive to paraffin-embedded, formalin-fixed, nonmalignant lung biopsies, the monoclonal antibody SCLC 5023 reacted with tumor cell infiltrates in biopsies from small cell lung carcinoma patients (14 of 14 cases positive), using the immunoperoxidase technique. This monoclonal reagent also reacted with other lung tumor cell types, including atypical carcinoid (5 of 5 positive), epidermoid (4 of 6 positive), undifferentiated and bronchoalveolar (3 of 4 cases each positive) carcinomas. By contrast, monoclonal antibody SCLC 2051 apparently identified an antigen expressed preferentially on small cell lung carcinoma cells (12 of 14 positive) and only rarely reacted with other lung tumor cell types (2 of 34 positive). Both monoclonal antibodies were negative to colon carcinoma, epidermoid carcinoma of the floor of the mouth, breast adenocarcinoma, and B- and T-cell leukemia and lymphoma cells, as determined by the enzyme-linked immunosorbent assay, indirect immunofluorescence, and immunoperoxidase techniques. These observations suggest that SCLC 2051 and SCLC 5023 may be of value in identifying tumor-associated antigens expressed in small cell and other lung carcinomas. In addition, the generation of antibody-producing cells towards common tumor-associated antigens may be enhanced by immunization with multiple tumor cell lines of the same histological type.
两种人小细胞肺癌细胞系NCI-H69和NCI-H128被用作交替免疫原来源，以产生针对小细胞肺癌相关抗原的单克隆抗体。用七次活肿瘤细胞注射使BALB/c小鼠致敏，四次注射NCI-H69细胞，三次注射NCI-H128细胞。使用来自SP2/0 Ag14细胞系的同基因骨髓瘤细胞，通过免疫小鼠脾细胞融合进行体细胞杂交。用酶联免疫吸附测定法针对小细胞肺癌细胞和正常肺成纤维细胞筛选杂交瘤集落。与仅用NCI-H69或NCI-H128细胞免疫的动物相比，交替免疫导致产生数量显著更多的与两种肿瘤细胞系均有选择性反应的杂交瘤。通过交替免疫产生的两个反应性杂交克隆SCLC 2051和SCLC 5023的单克隆抗体，对包括肺、肾、肝、脾、乳腺、甲状腺、脑、小肠和结肠在内的正常人体组织均呈阴性。虽然两种单克隆抗体对石蜡包埋、福尔马林固定的非恶性肺活检标本均无反应，但使用免疫过氧化物酶技术，单克隆抗体SCLC 5023与小细胞肺癌患者活检标本中的肿瘤细胞浸润有反应（14例中有14例阳性）。这种单克隆试剂也与其他肺肿瘤细胞类型有反应，包括非典型类癌（5例中有5例阳性）、表皮样癌（6例中有4例阳性）、未分化癌和细支气管肺泡癌（各4例中有3例阳性）。相比之下，单克隆抗体SCLC 2051显然识别出一种优先在小细胞肺癌细胞上表达的抗原（14例中有12例阳性），并且仅很少与其他肺肿瘤细胞类型有反应（34例中有2例阳性）。通过酶联免疫吸附测定法、间接免疫荧光法和免疫过氧化物酶技术测定，两种单克隆抗体对结肠癌、口底表皮样癌、乳腺腺癌以及B细胞和T细胞白血病及淋巴瘤细胞均呈阴性。这些观察结果表明，SCLC 2051和SCLC 5023在识别小细胞肺癌和其他肺癌中表达的肿瘤相关抗原方面可能有价值。此外，用相同组织学类型的多种肿瘤细胞系免疫可能会增强针对常见肿瘤相关抗原的抗体产生细胞的生成。
