Braathen L R
J Immunol Methods. 1980;35(1-2):43-56. doi: 10.1016/0022-1759(80)90149-0.
An anti-human T-lymphocyte antiserum has been further studied for specificity for T-cell subpopulations and application in tissue sections. Using a complement-dependent microcytotoxicity assay about 60% of normal peripheral blood T-cells were found to be sensitive to lysis, while 40% were resistant. When the T-cell suspensions were depleted of TG-cells using EA (Ripley) rosette sedimentation, the remaining cells demonstrated an increased percentage of lysis-sensitive cells, while the T-cells enriched in EA-RFC demonstrated a decreased percentage of cells sensitive to lysis, indicating that the antiserum was primarily directed against the non TG-cells, i.e. probably the TM helper cell population. This was further supported by functional studies. In order to quantitate T-cells, cytocentrifuge preparations were made from cell suspensions with known T-cell percentages and the T-cells determined with both the immune adherence technique using AET-treated sheep erythrocytes, as well as the indirect immunofluorescence technique using anti-T antiserum. The results of the two methods correlated well, suggesting that both methods can be used to determine T-cells in situ in infiltrate-like clusters of cells.
一种抗人T淋巴细胞抗血清针对T细胞亚群的特异性及其在组织切片中的应用得到了进一步研究。使用补体依赖的微量细胞毒性试验发现,约60%的正常外周血T细胞对裂解敏感,而40%具有抗性。当使用EA(里普利)玫瑰花结沉降法去除T细胞悬液中的TG细胞时,剩余细胞对裂解敏感的细胞百分比增加,而富含EA-RFC的T细胞对裂解敏感的细胞百分比降低,这表明该抗血清主要针对非TG细胞,即可能是TM辅助细胞群体。功能研究进一步支持了这一点。为了对T细胞进行定量,从已知T细胞百分比的细胞悬液中制备细胞离心涂片,并使用经AET处理的绵羊红细胞通过免疫黏附技术以及使用抗T抗血清的间接免疫荧光技术来测定T细胞。两种方法的结果相关性良好,表明这两种方法均可用于在浸润样细胞簇中原位测定T细胞。