摘要
为了制备抑制素α亚基（ＩＮＨα）单抗（ＭｃＡｂ）及研究ＩＮＨα在卵巢癌中的来源，以合成的人ＩＮＨα片段与载体蛋白（ＴＧ／ＢＳＡ）偶联复合物为抗原，采用杂交瘤技术，制备特异抗ＩＮＨα单抗，并用ＬＳＡＢ免疫组化染色程序，对人正常卵巢和卵巢癌中ＩＮＨα免疫活性进行定位。结果：①建立了３株分泌抗ＩＮＨαＭｃＡｂ的细胞株７Ａ２、１Ｇ６和１Ｇ１０；②观察到正常卵巢颗粒细胞和黄体细胞的胞浆内存在明显的强阳性着染，泡膜细胞呈弱阳性；卵巢癌细胞内也呈明显阳性，胞核内也有染色。结论：①抗ＩＮＨαＭｃＡｂ具有特异性，敏感性和实用性；
Purposes of this study were to prepare monoclonal antibodies (McAb) against human inhibin α subunit (INHα) and to study the sources of INHα produced in human ovarian malignant tumours and normal ovary.Immunogen was used with chemically synthetic N termical fragment INHα(1～26)Tyr·Gly conjugated with thyroglobulin(TG).The spleen cells from the BALB/C mouse immunized fused with NS 1 mouse myeloma cells.The hybridoma culture supernatants were screened by indirect ELISA caoted with the fragment coupled with bovine serum albumin(BSA).Immunological properties of McAb were analysed with recombinant human inhibin as a standard sample.Immunolocalization of INHα was determined on the paraffin imbeded tissue sections incubated with McAb.The visualization of the antigen antibody complex was carried out by the labelled strepoavidin biotin peroxidase(LSAB)procedure.Results were as fallows.Three recloned cell lines 7A2,1G6 and 1G10 were carefully selected.McAb titre in ascites fluid was up to 1.7×10 -5 。McAb did not cross react with TG or BSA,and human chorionic gonadotropin,follcle stimulating hormone,prolactin,oxytocin,vasopressin,estradiol,progesterone and testosterone.In the normal ovary immunoreactiivity of McAb was observed intensely in granulosa and luteal cells,weaklier in post menopausal ovary,and faintly in thecal cells,but not in other cells.Intense staining was observed in the tumour cells of human ovarian cancer including granulosa cell tumour,mucinous and serous cystadenocarcinomas.The positive staining was found largely in the cytoplasm,and partially in the cell nucleus.The negative staining was observed in all control sections It is concluded that:①The anti INHα McAbs have specifity,sensitivity and practicality.②Inhibin is produced by ovarian cancer cells,and used likely as a marker of postmenopausal ovarian cancers.
出处
《河南医科大学学报》
1998年第3期59-63,共5页
Journal of Henan Medical University
基金
卫生部青年科研基金
河南医科大学青年科研基金
关键词
抑制素
卵巢肿瘤
单克隆抗体
免疫定位
Α亚基
inhibin
sythetic peptide
ovarian neoplasms
monoclonal antibody
immunohistochemistry
hybridoma
