ANTIBODIES TO MOUSE LUNG CAPILLARY ENDOTHELIUM
ANTIBODIES TO MOUSE LUNG CAPILLARY ENDOTHELIUM
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DOI：
10.1177/36.7.3290332
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发表时间：
1988-07-01
影响因子：
3.2
通讯作者：
KENNEL, SJ
中科院分区：
文献类型：
--
作者：
RORVIK, MC;ALLISON, DP;KENNEL, SJ
我们有兴趣开发能够识别BALB/c小鼠肺部特定细胞类型的单克隆抗体（MoAb）。用正常小鼠肺组织匀浆免疫F344大鼠，用大鼠脾细胞与小鼠骨髓瘤SP 2/0融合制备杂交瘤细胞。选择产生在肺细胞免疫组织化学中有活性的MoAb的两个杂交瘤。MoAb 273- 34 A和411- 201 B在光镜水平均显示肺肺泡壁内毛细血管内皮细胞的广泛过氧化物酶染色。为了证明细胞特异性，用金标记抗体进行免疫电镜检查。将轻度固定的肺冷冻并薄切片，然后用MoAb和偶联至二抗的5 nm金颗粒染色。这些冷冻切片的定量分析表明，这两种抗体，在最佳浓度下使用，是特异性结合毛细血管内皮细胞。超过95%的金颗粒与肺泡壁薄侧的毛细血管内皮细胞相关。当毛细血管毗邻含有间质细胞的厚隔膜时，约三分之二的金颗粒与内皮细胞结合，约四分之一与间质细胞结合。这些单克隆抗体将有助于研究内皮细胞在中毒性肺损伤中的作用。
We are interested in developing monoclonal antibodies (MoAbs) that recognize specific cell types in the lung of BALB/c mice. Normal mouse lung homogenate was used to immunize F344 rats and hybridomas were produded by fusion of rat spleen cells with mouse myeloma SP 2/0. Two hybridomas were selected which produced MoAbs active in immunohistochemistry of lung cells. MoAb 273-34A and 411-201B both show extensive peroxidase staining of capillary endothelial cells within alveolar walls of lungs at the light microscopic level. To demonstrate cell specificity, immunoelectron microscopy with gold-labeled antibody was performed. Lightly fixed lungs were frozen and thinsectioned before staining with MoAb and 5-nm gold particles coupled to secondary antibody. Quantitative analyses of these cryosections show that both antibodies, used at optimal concentrations, are specific for binding to capillary endothelial cells. More than 95% of the gold particles are associated with capillary endothelial cells on the thin side of the alveolar wall. When capillaries adjoined thick septa containing interstitial cells, about two thirds of the gold particles were associated with endothelial cells and about one quarter with interestitial cells. These MoAbs should be useful in studying the role of endothelial cells in toxic lung injury.