Wong J H, Xu S H, Saxton R E, Gupta R K, Morton D L
John Wayne Cancer Clinic, UCLA School of Medicine 90024.
Int J Cancer. 1990 Mar 15;45(3):445-9. doi: 10.1002/ijc.2910450312.
Detection of antigen-specific circulating immune complexes (IC) in sera from cancer patients provides an approach for defining tumor antigens that induce a host immune response and could prove useful for purposes of immunoprognosis. In the present study, a sandwich ELISA was developed to detect antigen-specific IC in cancer patients, utilizing a murine monoclonal antibody designated MAb JSI. The MAb was produced by cell fusion using standard hybridoma technology following immunization with a partially purified fetal antigen that had been isolated from the spent culture medium of a melanoma cell line. The partially purified antigen appeared to be broadly expressed on melanomas, sarcomas, and carcinomas. The MAb was mass-produced in pristine-primed mice. MAb JSI reacted with the cultured melanoma cell M14 but not the autologous lymphoblastoid cell L14. Following purification by ammonium sulfate precipitation, the MAb was immobilized on polystyrene plates and utilized to capture antigen-specific immune complexes from sera of melanoma patients which were detected with anti-human globulins. The second antibody in the sandwich was shown to be endogenous human IgG. Antigen-specific immune complexes were present in melanoma patients but only infrequently in sera from normal individuals and patients with active autoimmune disease. Antigen-specific immune complexes detected in melanoma sera were isolated by affinity chromatography utilizing the MAb JSI. The assay described in this report is simple and reproducible, without plate effect (p = 0.97) or time effect (p = 0.34) and could provide a useful new approach to examine the role of antigen-specific circulating immune complex analysis in cancer patients.
检测癌症患者血清中的抗原特异性循环免疫复合物（IC）为定义诱导宿主免疫反应的肿瘤抗原提供了一种方法，并且可能对免疫预后具有重要意义。在本研究中，开发了一种夹心ELISA法来检测癌症患者中的抗原特异性IC，该方法使用一种名为单克隆抗体JSI的鼠单克隆抗体。该单克隆抗体是在用从黑色素瘤细胞系的废培养基中分离出的部分纯化胎儿抗原免疫后，采用标准杂交瘤技术通过细胞融合产生的。该部分纯化抗原似乎在黑色素瘤、肉瘤和癌组织中广泛表达。该单克隆抗体在经原始启动的小鼠中大量生产。单克隆抗体JSI与培养的黑色素瘤细胞M14反应，但不与自体淋巴母细胞L14反应。通过硫酸铵沉淀纯化后，将该单克隆抗体固定在聚苯乙烯板上，并用于从黑色素瘤患者血清中捕获抗原特异性免疫复合物，然后用抗人球蛋白进行检测。夹心ELISA中的二抗显示为内源性人IgG。抗原特异性免疫复合物存在于黑色素瘤患者中，但在正常个体和活动性自身免疫性疾病患者的血清中很少见。利用单克隆抗体JSI通过亲和层析从黑色素瘤血清中分离出抗原特异性免疫复合物。本报告中描述的检测方法简单且可重复，无板效应（p = 0.97）或时间效应（p = 0.34），可为研究抗原特异性循环免疫复合物分析在癌症患者中的作用提供一种有用的新方法。
