He X M, Wikstrand C J, Friedman H S, Bigner S H, Pleasure S, Trojanowski J Q, Bigner D D
Department of Pathology, Duke University Medical Center, Durham, North Carolina.
Lab Invest. 1991 Jun;64(6):833-43.
Three new human medulloblastoma (MB) cell lines (D384 Med, D425 Med, and D458 Med) and their transplantable xenografts were examined for antigenic expression with antibodies against neuroectodermal antigens, cytoskeletal proteins, neuroendocrine markers, glioma-associated antigens, tenascin, human lymphocyte antigen molecules, epidermal growth factor receptor, and T-cell antigen by indirect immunofluorescence, avidin-biotin complex peroxidase immunohistochemistry, and immunoblot methods. We found that each of the three cell lines expressed vimentin; low-, middle-, and high-molecular-weight neurofilament proteins; and the synaptic vesicle membrane glycoprotein synaptophysin. Each of the cell lines also reacted with antibodies against neural cell adhesion molecules, but none of them were positive for antibodies against glial fibrillary acidic protein, keratin, microtubule-associated protein tau and microtubule-associated protein 2, human lymphocyte antigen-DR, epidermal growth factor receptor, and T-cell antigen. Immunoreactivities with anti-tenascin and anti-glioma-associated antibodies were variable in these cell lines. Anti-human lymphocyte antigen-A,B and anti-beta 2-microglobulin antibodies reacted with xenografts of D384 Med and D425 Med and were weakly positive for a small population of D384 Med cultured cells. In summary, the detection of neurofilament proteins and synaptophysin and the absence of glial fibrillary acidic protein provide strong evidence for a neuronal phenotype of D384 Med, D425 Med, and D458 Med.
利用抗神经外胚层抗原、细胞骨架蛋白、神经内分泌标志物、胶质瘤相关抗原、腱生蛋白、人类淋巴细胞抗原分子、表皮生长因子受体及T细胞抗原的抗体,通过间接免疫荧光、抗生物素蛋白-生物素复合物过氧化物酶免疫组织化学及免疫印迹法,对三种新的人类髓母细胞瘤(MB)细胞系(D384 Med、D425 Med和D458 Med)及其可移植异种移植物进行了抗原表达检测。我们发现,这三种细胞系均表达波形蛋白、低、中、高分子量神经丝蛋白以及突触囊泡膜糖蛋白突触素。每种细胞系还与抗神经细胞黏附分子抗体发生反应,但均未对抗胶质纤维酸性蛋白、角蛋白、微管相关蛋白tau和微管相关蛋白2、人类淋巴细胞抗原-DR、表皮生长因子受体及T细胞抗原的抗体呈阳性反应。在这些细胞系中,与抗腱生蛋白和抗胶质瘤相关抗体的免疫反应性各不相同。抗人类淋巴细胞抗原-A、B及抗β2-微球蛋白抗体与D384 Med和D425 Med的异种移植物发生反应,且对一小部分D384 Med培养细胞呈弱阳性。总之,神经丝蛋白和突触素的检测以及胶质纤维酸性蛋白的缺失为D384 Med、D425 Med和D458 Med的神经元表型提供了有力证据。