Kasinrerk W, Tokrasinwit N, Naveewongpanit P
Department of Clinical Immunology, Faculty of Associated Medical Sciences, Chiang Mai University, Thailand.
J Med Assoc Thai. 1998 Nov;81(11):879-92.
A hybridoma secreting monoclonal antibody (mAb) specific to CD4 protein was generated. This monoclonal antibody, named MT4, was proved to be specific to CD4 protein as it reacted with CD4-DNA transfected COS cells, CD4+ cell lines and CD4+ lymphocytes. Furthermore, MT4 mAb inhibited the binding of standard CD4 monoclonal antibodies to CD4 proteins on CD4+ cells. To develop a home made reagent for CD4+ lymphocyte determination by flow cytometry, fluorescein isothiocyanate (FITC) was conjugated to MT4 mAb. To evaluate the developed reagent, 30 HIV infected and 30 healthy individuals were determined for CD4+ lymphocytes by using both a commercial Simultest reagent kit and home made FITC labeled MT4 mAb simultaneously. The study has shown that both percentages and absolute CD4+ lymphocyte counts obtained from both reagents were equivalent. The correlation coefficient for regression analysis was 0.995 and 0.996 for percentages and absolute CD4+ lymphocyte counts, respectively. The results suggest that home made FITC labeled MT4 reagent is an acceptable alternative reagent for monitoring CD4+ lymphocytes in blood samples by flow cytometry.
产生了一种分泌针对CD4蛋白的单克隆抗体（mAb）的杂交瘤。这种名为MT4的单克隆抗体被证明对CD4蛋白具有特异性，因为它能与转染了CD4-DNA的COS细胞、CD4+细胞系和CD4+淋巴细胞发生反应。此外，MT4单克隆抗体抑制了标准CD4单克隆抗体与CD4+细胞上CD4蛋白的结合。为了开发一种用于通过流式细胞术检测CD4+淋巴细胞的国产试剂，将异硫氰酸荧光素（FITC）与MT4单克隆抗体偶联。为了评估所开发的试剂，同时使用商业Simultest试剂试剂盒和国产FITC标记的MT4单克隆抗体对30名HIV感染者和30名健康个体的CD4+淋巴细胞进行检测。研究表明，两种试剂所获得的CD4+淋巴细胞百分比和绝对计数均相当。百分比和绝对CD4+淋巴细胞计数的回归分析相关系数分别为0.995和0.996。结果表明，国产FITC标记的MT4试剂是通过流式细胞术监测血样中CD4+淋巴细胞的一种可接受的替代试剂。
