De Waele M, De Mey J, Moeremans M, Smet L, Broodtaerts L, Van Camp B
J Histochem Cytochem. 1983 Apr;31(4):471-8. doi: 10.1177/31.4.6186735.
Immunogold staining in combination with enzyme cytochemistry was used to determine the cytochemical profile of the immunoregulatory T-lymphocyte subpopulations defined by the monoclonal antibodies OKT3, OKT4, OKT8, and OKM1 in normal peripheral blood. Leukocyte suspensions were first incubated with the monoclonal mouse antibodies and then with colloidal gold-labeled goat antimouse antibodies. The cells were fixed and cytocentrifuge preparations were made. Cytochemical reactions for the detection of peroxidase, acid alpha-naphthyl acetate esterase, acid phosphatase, and beta-glucuronidase were performed on these preparations. Under light microscopy, lymphocytes reacting with the monoclonal antibodies had numerous dark granules around their surface membrane. In the cytoplasm the intracellular enzymatic activities were stained. The T-lymphocytes were characterized by a dot-like activity for the three enzymes. No significant difference could be found between the cytochemical profile of the T-helper (OKT4 positive) and T-cytotoxic suppressor cell populations (OKT8 positive). A few cells with lymphocyte morphology reacted with the OKM1 monoclonal antibody. Their cytochemical characteristics were different from those of mature T-cells (OKT3 population) or mature monocytes. From the comparison of their cytochemical characteristics, we can conclude that there is little correlation between the immunoregulatory T-lymphocyte subsets defined by these monoclonal antibodies and those defined by Fc receptors.
免疫金染色结合酶细胞化学法用于确定正常外周血中由单克隆抗体OKT3、OKT4、OKT8和OKM1所定义的免疫调节性T淋巴细胞亚群的细胞化学特征。首先将白细胞悬液与小鼠单克隆抗体孵育,然后与胶体金标记的山羊抗小鼠抗体孵育。细胞固定后制成细胞离心涂片。对这些涂片进行检测过氧化物酶、酸性α-萘乙酸酯酶、酸性磷酸酶和β-葡萄糖醛酸酶的细胞化学反应。在光学显微镜下,与单克隆抗体反应的淋巴细胞在其表面膜周围有许多深色颗粒。在细胞质中,细胞内酶活性被染色。T淋巴细胞的特征是这三种酶呈现点状活性。T辅助细胞(OKT4阳性)和T细胞毒性抑制细胞群体(OKT8阳性)的细胞化学特征之间未发现显著差异。少数具有淋巴细胞形态的细胞与OKM1单克隆抗体反应。它们的细胞化学特征与成熟T细胞(OKT3群体)或成熟单核细胞不同。通过比较它们的细胞化学特征,我们可以得出结论,这些单克隆抗体所定义的免疫调节性T淋巴细胞亚群与由Fc受体所定义的亚群之间几乎没有相关性。