Human monoclonal antibodies reactive with colon carcinoma: identification by a novel screening procedure with xenografts.
Human monoclonal antibodies reactive with colon carcinoma: identification by a novel screening procedure with xenografts.
复制标题
与结肠癌反应的人单克隆抗体：通过异种移植物的新型筛选程序进行鉴定。
DOI：
10.1002/jcla.1860030109
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发表时间：
1989
影响因子：
2.7
通讯作者：
Mitchell,MS
中科院分区：
文献类型：
--
作者：
Kan-Mitchell,J;Formenti,SC;Mitchell,MS
为了鉴定对人具有免疫原性的肿瘤相关抗原，通过将三名结肠腺癌患者的局部淋巴结淋巴细胞（LNL）与M5小鼠骨髓瘤细胞系融合来产生人单克隆抗体（MAb）。19个威尔斯孔中的异源杂交瘤分泌人免疫球蛋白（lg）至少60天。为了鉴定免疫反应性单克隆抗体，我们设计了一种免疫组织化学测定与异种移植的六个结肠肿瘤。用生物素缀合的山羊抗人lg抗体，然后用抗生物素蛋白-生物素-过氧化物酶复合物观察结合。该试验检测结合的灵敏度为0.5 ug/ml人Ig。在19份上清液中的14份中检测到结肠异种移植物冷冻切片中的肿瘤相关抗原（TAA）反应性。然而，只有9个（3个IgG、5个IgM和1个伊加）对福尔马林固定切片具有反应性，表明福尔马林固定破坏了部分但非全部抗原性。克隆了四种MAb，并针对整个19种异种移植物组进行了测试。一种IgM单克隆抗体仅对结肠癌反应，而其他三种（伊加、IgG和IgM各一种）具有更广泛的反应性。四种人单克隆抗体中有三种与黑素瘤没有交叉反应。总之，人局部淋巴结含有高比例的对自体肿瘤致敏的B淋巴细胞。人结肠肿瘤异种移植物的免疫染色可以容易地鉴定与结肠癌反应的人MAb。
To identify tumor‐associated antigens immunogenic to man, human monoclonal antibodies (MAbs) were generated by fusing regional lymph node lymphocytes (LNLs) of each of three colon adenocarcinoma patients with the M5 mouse myeloma cell line. Heterohybridomas in 19 wells secreted human immunoglobulin (lg) for at least 60 days. To identify immunoreactive MAbs, we devised an immunohistochemical assay with xenografts of six colon tumors. Binding was visualized with a biotin‐conjugated goat antihuman lg antibody followed by an avidin‐biotin‐peroxidase complex. This assay detected binding with a sensitivity of 0.5 ug/ml human lg. Reactivity against tumor‐associated antigens (TAAs) in frozen sections of the colon xenografts was detected in 14 of the 19 supernates. However, only nine (three IgGs, five lgMs and one IgA) were reactive against formalin‐fixed sections, showing that formalin‐fixation destroyed some but not all antigenicity. Four MAbs were cloned and tested against the entire panel of 19 xenografts. One IgM MAb reacted only against colon carcinomas, while the other three (one each of IgA, IgG and IgM) had broader reactivities. Three of the four human MAbs did not cross‐react with melanomas. In summary, human regional lymph nodes contain a high proportion of B lymphocytes sensitized to the autochthonous tumor. Immunostaining of human colon tumor xenografts can readily identify human MAbs reactive with colon carcinomas.