モノクローナル抗体による癌・トロホブラスト抗原の検討

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In order to investigate oncotrophoblast antigens, 8 monoclonal antibodies(MoAbs) which react with normal and/or neoplastic human trophoblasts have been produced,and examined their reactivities with other non-trophoblast origin tumors. In addition, several MoAbs have been studied for their affinities to the tumor in vivo. Immunogens of the 6 MoAbs(TM7-3,TM3-8,TM5-1,TM8-13 TM4-5 and TM3-3) were cultured choriocarcinoma cell lines,and those of the other 2 MoAbs(M-rp12,andM-rp 28) were vH-ras oncogene product P21. BALB-c mice were immunized by ip or sc injection with (A) cultured human choriocarcinoma cell lines, BeWo,NUC-1,or JAR respectively, or (B) recombinant P21 produced by Escherichia coli. Spleen cells of the immunized mice were fused with mouse myeloma cells with minor modifications according to the technique reported by Kohler and Milstein. To isolate hybridoma cell lines producing antibody specific for (A) normal and-or neoplastic trophoblasts, or (B) v-H-ras oncogene product P21,screening of hybridomas and cloning were performed. Ig isotypes of TM7-3,TM3-8,TM5-1,TM8-13 and TM3-3 are IgG1,TM4-5 is IgG2a and,M-rp12 and M-rp28 are IgM. According to the distributions of the antigens detected by the MoAbs,TM7-3 and TM3-8,TM4-5 and TM3-3,and M-rp12 and M-rp28 showed a similar reactive pattern respectively. Among these MoAbs, both TM7-3 and TM3-8 showed an unique reactive pattern. They demonstrated a very strong reaction with choriocarcinoma,particularly cytotrophoblast-like tumor cells,in spite of a very weak reaction in normal chorionic villi. In addition,they showed a positive reaction with ovarian serous cysy adenocarcinoma(3/4) and well differentiated uterine corpus carcinoma(2/2). Scintigraphy performed 6 days after injection of 131^I-labelled TM3-8 to choriocarcinoma(BeWo) transplanted nude mice, demonstrated the excellent visualization of the tumor. Radioactivities of the tumor and other normal tissues at 6th days from the injection,showed a selective binding of TM3-8 to the tumor. Whole body autoradiography of mouse was performed 4 days after injection of 125^I-labelled TM3-8 to the mice with a choriocarcinoma. The autoradiogram showed a clear positive gains restricted to the tumor. In contrast with TM7-3 and TM3-8,TM5-1 demonstrated a positive reaction not in cytotrophoblast but in syncytiotrophoblast selectively regardless of normal chorionic villi or choriocarcinoma. With non-trophoblast origin tumor,TM5-1 showed no positive reaction. A positive reaction of TM8-13 was demonstrable not in villous trophoblast but in partial non-villous trophoblast,and kindney carcinoma(2/6). TM4-5 and TM3-3 were MoAbs reacting to normal and molar trophoblasts. They gave a weak reaction to choriocarcinoma. Among non-trophoblast origin tumors, a positive reaction was observed clearly in ovarian carcinoma. A positive reaction of M-rp12 and M-rp28 was not seen in normal and molar trophoblasts,but they reacted to choriocarcinoma. Further,they reacted to uterine cervical carcinoma, endometrial carcinoma and ovarian carcinoma among non-trophoblast origin tumors. All MoAbs in the present studies are related to trophoblast or choriocarcinoma basically. They react,however,to some kinds of non-trophoblast origin tumors in various pattern respectively. These findings are consistent with earlier reports of cancer cell's phenotypic change to trophoblast. In addition,with some of the MoAbs,encouraging data was obtained for thinking of a possibility of the practical application such as imaging of tumors in clinical field.
社団法人日本産科婦人科学会の論文
1987-08-01

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