Tabibzadeh S S, Gerber M A
J Immunol Methods. 1986 Jul 24;91(2):169-74. doi: 10.1016/0022-1759(86)90475-8.
The development of monoclonal antibodies has allowed characterization of subpopulations of lymphoid cells and of their in situ distribution in tissues. The feasibility of simultaneous localization of two surface antigens was studied by double staining with monoclonal antibodies to B cells, T cell subsets and follicular dendritic reticulum cells (DRC) using the avidin-biotin-complex (ABC) method in sections of human lymphoid tissues (tonsil, lymph node, spleen) and a non-lymphoid tissue, endometrium. Color mixture was avoided when an additional incubation with avidin-biotin-labeled peroxidase and subsequent development in the respective substrate of the first sequence preceded the second staining sequence using the primary antibodies at optimal concentrations. The antigenic profiles were portrayed by contrasting and distinct colors of the reaction products. It was observed that T lymphocytes of the cytotoxic/suppressor and helper/inducer phenotypes were topographically associated with each other and with B cells in B and T cell areas of lymphoid tissues as well as in lymphocytic aggregates in endometria. Subpopulations of these cells were mantled by processes of DRCs in lymphoid follicles. The findings indicate that the double ABC staining method can be used for simultaneous demonstration of two surface antigens in tissue sections.
单克隆抗体的发展使得对淋巴样细胞亚群及其在组织中的原位分布进行特征描述成为可能。采用抗生物素蛋白-生物素复合物（ABC）法，利用针对B细胞、T细胞亚群和滤泡树突状网状细胞（DRC）的单克隆抗体，对人淋巴样组织（扁桃体、淋巴结、脾脏）和一种非淋巴样组织子宫内膜的切片进行双重染色，研究同时定位两种表面抗原的可行性。当在使用最佳浓度的一抗进行第二次染色序列之前，先用抗生物素蛋白-生物素标记的过氧化物酶进行额外孵育，然后在第一个序列的相应底物中显色时，避免了颜色混合。通过反应产物对比鲜明且独特的颜色描绘抗原图谱。观察到，具有细胞毒性/抑制和辅助/诱导表型的T淋巴细胞在淋巴样组织的B和T细胞区域以及子宫内膜的淋巴细胞聚集体中，在地形学上彼此相关且与B细胞相关。这些细胞的亚群被淋巴滤泡中的DRC突起所覆盖。研究结果表明，双重ABC染色法可用于在组织切片中同时显示两种表面抗原。
