Monoclonal antibodies against rat Leydig cell surface antigens.
Monoclonal antibodies against rat Leydig cell surface antigens.
复制标题
针对大鼠 Leydig 细胞表面抗原的单克隆抗体。
DOI：
--
复制
发表时间：
1986
影响因子：
3.6
通讯作者：
Edward M. Eddy
中科院分区：
文献类型：
--
作者：
M. Hedger;Edward M. Eddy
针对间质细胞表面的单克隆抗体（mab）可用于睾丸制剂中该细胞的鉴定。采用Percoll密度梯度对胶原酶分散的成年大鼠间质细胞进行分离，利用间质细胞富集部分制备单克隆抗体。采用酶联免疫吸附法（ELISA）、间接免疫荧光法（IIF）对离体睾丸细胞进行筛选，并对成年睾丸石蜡切片进行免疫细胞化学定位。在密度梯度分数中，免疫球蛋白（Ig） M单克隆抗体（LC-1C6和LC-6H6）标记具有间质细胞形态特征的细胞表面。IIF和ELISA观察到的单抗结合活性的密度梯度曲线与[125I]-人绒毛膜促性腺激素（hCG）结合、体外睾酮对hCG的反应、3 β -羟基类固醇脱氢酶组织化学和电镜观察到的间质细胞分布相似。单克隆抗体在切片中显著标记了大多数间质细胞，但结缔组织、内皮细胞或精小管细胞很少或没有标记。这两种单克隆抗体都在富含Leydig细胞提取物的Western印迹中识别Mr 58000的成分。结果表明，LC-1C6和LC-6H6能够识别Leydig细胞表面的抗原，而这些抗原不存在于其他离体大鼠睾丸细胞上。这些单克隆抗体是间质细胞在原位和体外的特异性标记物。
Monoclonal antibodies (MAbs) directed against the Leydig cell surface may be used to identify this cell in testicular preparations. Collagenase-dispersed adult rat interstitial cells were fractionated on Percoll density gradients, and Leydig cell-enriched fractions were used to prepare MAbs. Hybridomas were screened by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IIF) on isolated testicular cells and immunocytochemical localization on paraffin sections of adult testes. In density gradient fractions, immunoglobulin (Ig) M MAbs (LC-1C6 and LC-6H6) labeled the surface of cells possessing the morphological characteristics of Leydig cells. The density gradient profiles of MAb-binding activity observed by IIF and ELISA were parallel with the Leydig cell distribution as determined by [125I]-human chorionic gonadotropin (hCG) binding, testosterone response to hCG in vitro, 3 beta-hydroxysteroid dehydrogenase histochemistry and electron microscopy. The MAbs prominently labeled most interstitial cells in sections, but there was little or no labeling of connective tissue, endothelial or seminiferous tubule cells. Both MAbs recognized components of Mr 58,000 in Western blots of Leydig cell-enriched extracts. The results indicate that LC-1C6 and LC-6H6 recognize antigens on the Leydig cell surface that are not present on other isolated testicular cells from the adult rat. These MAbs are specific markers of the Leydig cell in situ and in vitro.