Ebihara T, Sakai N, Koyama S
Department of Internal Medicine, University of Tsukuba, Japan.
Tohoku J Exp Med. 1990 Nov;162(3):213-24. doi: 10.1620/tjem.162.213.
Mononuclear peripheral blood lymphocytes (PBL) from patient with infectious mononucleosis (IM) were tested in a 51Cr-release assay for cytotoxicity against autologous and allogeneic lymphoblastoid cell line (LCL), or Epstein-Barr virus (EBV)-genome positive and negative cell line. In acute phase, PBL lyse an autologous LCL as well as allogeneic LCL (Wa cells). High levels of cytotoxicity were observed in the combinations between effector and target cells sharing HLA-Class 1 product. EBV-genome positive Daudi and Raji cells which lack HLA-Class 1 antigen and have mismactched HLA-Class 1 antigen, respectively showed resistance to killing. EBV-genome negative tumor cells except NK sensitive K562 cells were not killed by IM lymphocytes. However, the IM lymphocytes without atypical form in convalescent phase failed to show killing activity against autologous and allogeneic LCL. These findings suggest that cell surface membrane antigen structure on EBV-infected LCL may be able to explain the recognition and triggering of lysis of target cells by HLA-Class 1 restricted cytotoxic T cells (CTL) from acute IM. Phenotypic analysis of PBL with atypical form from IM was made by two-color flow cytometry. The data demonstrate that CD8+ T cells quantitatively represent the major population of lymphocytes expanded during acute IM. Furthermore, approximately 70% of these CD8+ T cells express HLA-DR on these surface, suggesting that they have undergone activation. However, IL 2R (CD25 antigen) expression was not significantly elevated on activated T cells. The salient profile on cytofluorographs of an acute IM was the increased number of CD3+CD19-, CD8+CD11b-, CD8+CD28+ and CD8+S6F1+ cells. However, CD3-CD19+, CD8+CD11b+, CD8+S6F1-, CD4+Leu8- and CD25+HLA-DR+ antigens were little expressed. Increased number of CD8+CD11b-, CD8+CD28+ and CD8+S6F1+ cells, which are regarded as CTL were reduced according to the improvement of the clinical symptoms and laboratory findings. These results together with HLA typing analysis suggested a possibility HLA-Class 1 restriction of the CTL with surface phenotype of CD8+CD11b-, CD8+CD28+, and CD8+S6F1+.
采用51Cr释放试验，检测传染性单核细胞增多症（IM）患者的单核外周血淋巴细胞（PBL）对自体和异体淋巴母细胞系（LCL）或爱泼斯坦-巴尔病毒（EBV）基因组阳性和阴性细胞系的细胞毒性。在急性期，PBL可裂解自体LCL以及异体LCL（Wa细胞）。在效应细胞和靶细胞共享HLA-Ⅰ类产物的组合中观察到高水平的细胞毒性。EBV基因组阳性的Daudi细胞和Raji细胞分别缺乏HLA-Ⅰ类抗原和具有不匹配的HLA-Ⅰ类抗原，对杀伤均表现出抗性。除NK敏感的K562细胞外，EBV基因组阴性肿瘤细胞未被IM淋巴细胞杀伤。然而，恢复期无异常形态的IM淋巴细胞对自体和异体LCL未表现出杀伤活性。这些发现表明，EBV感染的LCL上的细胞表面膜抗原结构可能能够解释急性IM中HLA-Ⅰ类限制性细胞毒性T细胞（CTL）对靶细胞的识别和裂解触发。通过双色流式细胞术对IM患者有异常形态的PBL进行表型分析。数据表明，CD8+T细胞在数量上代表急性IM期间扩增的淋巴细胞的主要群体。此外，这些CD8+T细胞中约70%在其表面表达HLA-DR，表明它们已经经历了激活。然而，活化T细胞上IL-2R（CD25抗原）的表达并未显著升高。急性IM的细胞荧光图上的显著特征是CD3+CD19-、CD8+CD11b-、CD8+CD28+和CD8+S6F1+细胞数量增加。然而，CD3-CD19+、CD8+CD11b+、CD8+S6F1-、CD4+Leu8-和CD25+HLA-DR+抗原表达很少。随着临床症状和实验室检查结果的改善，被视为CTL的CD8+CD11b-、CD8+CD28+和CD8+S6F1+细胞数量减少。这些结果与HLA分型分析一起提示，具有CD8+CD11b-、CD8+CD28+和CD8+S6F1+表面表型的CTL可能存在HLA-Ⅰ类限制。
