抗人肝癌细胞膜单克隆抗体HCMP－1的制备及其生物学特性的初步鉴定  

PRODUCTION AND CHARACTERIZATION OF MOUSE MONOCLONAL ANTIBODY DIRECTED AGAINST HUMAN HEPATOCELLULAR CARCINOMA

作　　者：魏超[1] 汤钊猷[1] 刘康达[1] 林芷英[1] 韩根娣 

机构地区：[1]上海医科大学肝癌研究所

出　　处：《肿瘤》1994年第4期193-196,共4页Tumor

摘　　要：用超选择免疫法，诱导ＢＡＬＢ／ｃ小鼠对正常人肝细胞抗原产生选择性低免疫反应后，二步腹腔注射人肝癌细胞悬液，２～４周后用人肝癌细胞作强化免疫动物１次，取出脾细胞与ＳＰ２／０骨髓瘤细胞用ＰＥＧ法融合制备杂交瘤。采用细胞抗原ＥＬＩＳＡ法，同时测定培养上清对肝癌细胞及正常肝细胞的免疫反应，筛出一株能稳定分泌抗人肝癌细胞膜抗原的单克隆抗体（ＨＣＭＰ－１ＭＡｂ）的杂交瘤细胞株。ＨＣＭＰ－１ＭＡｂ是一种ＩｇＧ２ａ亚型的单抗，它与ＨＢｓＡｇ、ＨＢｃＡｇ、ＨＢｅＡｇ等乙型肝炎病毒相关抗原无交叉免疫反应，与ＡＦＰ－亦无阳性反应。免疫萤光法证实ＨＣＭＰ－１ＭＡｂ对ＱＧＹ－７７０３、ＢＥＬ－７４０２及ＳＭＭＣ－７７２１等人肝癌细胞株呈阳性反应。改良ＡＢＣ免疫组织化学证明ＨＣＭＰ－１ＭＡｂ与肝细胞癌组织呈阳性反应。说明ＨＣＭＰ－１ＭＡｂ对肝癌反应的阳性率较高，特导性较好。超选择免疫法，先联合应用免疫抑制药物使动物对正常肝细胞抗原产生低免疫反应性。再以肝癌细胞免疫时，有利于动物免疫系统对肿瘤相关抗原的识别及特异性抗体的产生。从而提高了杂交瘤制备的成功率。我们认为这一方法在单抗制备方面有较高的应用价值。In the present study,the high selected immunization project was used in an attempt to elicit host low immunoresponses to normal surface markers expressed on hepatocellular carcinoma cells. Cyclophosphamide was given to BaLB/c mice 3 days after the first injection of normal human liver cells,the mice were then given two additional immunizations with hepatocellular carcinoma (HCC) cells at 2-week intervals.3 days after the last immunization spleen cells were fused with myeloma SP2/0 cells. One hybridoma HCMP-1,producing monoclonal antibody against HCC cell membrane,was established.The amounts of chromosomes of the hybridoma are 96～106.McAb HCMP-1 is a kind of IgG2a isotype. Indirect membrane immunofluoresence assays demonstrated that McAb HCMP-1 reacted with 3 hepatocellular carcinoma cell lines,but not with K-562 cell line and normal human lymphocytes. McAb HCMP-1 was also positive in 7 of 8 HCC tissues but negative in other human tumor tissues and in 3 normal human liver tissues tested by ABC technique. This result suggests that McAb HCMP-1 may be used in the diagnostic and treatment of HCC.

关 键 词：肝细胞癌 单克隆抗体 杂交瘤 

分 类 号：R735.703[医药卫生—肿瘤]