Dillman S L, Aldenderfer P, Strelkauskas A
Dept. of Microbiology and Immunology, Medical University of South Carolina, Charleston 29425-2230.
Hum Antibodies Hybridomas. 1994;5(1-2):32-40.
A series of human hybridomas were derived by the fusion of the immature T cell line, JM, and lymphocytes from the peripheral blood of a colon carcinoma patient in long term remission who was shown to produce anti-tumor antibody. Five of the hybridomas have been selected for further study. These hybridomas express the T cell surface markers CD2, CD3, CD4, and CD8. The human IgG2/kappa antibody produced by these clones has been purified using affinity chromatography and has been used in immunohistiologic staining procedures. Biotinylated monoclonal antibody (MAb) stained colon carcinoma tissue but did not stain normal colon. Positive immunostaining was also seen utilizing colon carcinoma cell lines but not with other cell lines tested. These hybrids were constructed without using the conventional fusion technology which employs 8-azoguanine resistant, HAT sensitive malignant fusion partners. Because the fusion partner was a T cell, we were able to select hybrids on the basis of surface immunoglobulin. This approach was accompanied by stringent selection of patients as lymphocyte donors. Utilizing these unique methods, we have successfully produced hybridomas with T cell surface markers that produce human MAb with reactivity to colon carcinoma.
通过将未成熟T细胞系JM与一名处于长期缓解期的结肠癌患者外周血淋巴细胞进行融合,获得了一系列人杂交瘤,该患者被证明能产生抗肿瘤抗体。已挑选出5个杂交瘤进行进一步研究。这些杂交瘤表达T细胞表面标志物CD2、CD3、CD4和CD8。这些克隆产生的人IgG2/κ抗体已通过亲和层析法纯化,并已用于免疫组织学染色程序。生物素化单克隆抗体(MAb)可对结肠癌组织进行染色,但不能对正常结肠进行染色。利用结肠癌细胞系也观察到了阳性免疫染色,但对其他测试的细胞系则未观察到。这些杂交瘤的构建未使用传统的融合技术,传统技术采用对8-氮鸟嘌呤有抗性、对HAT敏感的恶性融合伙伴。由于融合伙伴是T细胞,我们能够根据表面免疫球蛋白选择杂交瘤。这种方法伴随着对作为淋巴细胞供体的患者进行严格挑选。利用这些独特的方法,我们成功地产生了具有T细胞表面标志物的杂交瘤,这些杂交瘤产生对结肠癌有反应性的人单克隆抗体。