Induction of human cytotoxic T cells by peptide-pulsed universal antigen presenting cells

我们检测了EB病毒（EBV）特异性细胞毒T细胞的存在、EBV特异性ctl的诱导以及诱导ctl在慢性活动性EBV感染（CAEBV）患者外周血CD8+细胞中的细胞毒活性，结果如下：1。通过hlaa24特异性和EBV特异性四聚体分析，在健康志愿者和传染性单核细胞增多症患者的外周血淋巴细胞中发现了EBV特异性ctl。有趣的是，我们还能够在CAEBV.2患者的外周血中证明BRLF1肽特异性ctl。诱导抗原特异性ctl：我们使用HLAA24阳性的自体B淋巴样细胞（B- lcls）作为抗原提呈细胞，用B- lcls刺激外周单个核细胞，用或不含BRLF1肽脉冲。即使在缺乏BRLF1特异性肽的情况下，我们也成功地诱导了BRLF1四聚体阳性的ctl。这表明EBV感染的b - lclc在没有进一步肽脉冲的情况下给予特异性ctl足够多的BRLF1特异性刺激。在健康志愿者和CAEBV患者的单核细胞中，观察到这些CD8阳性BRLF1四聚体特异性诱导和增殖ctl。利用ctl产生干扰素γ的能力对诱导ctl的细胞毒活性进行检测。首先，我们证实来自健康志愿者的BRLF1四聚体特异性CD8+ ctl仅在BRLF1特异性肽后才能产生干扰素γ。我们发现来自CAEBV患者的单个核细胞在受到特定肽刺激后也产生BRLF1四聚体特异性ctl。然而，来自患者的BRLF1特异性ctl仅产生约30%的干扰素γ，而来自健康志愿者的ctl产生约50%的干扰素γ。这些结果表明，来自健康志愿者和CAEBV患者的单个核细胞可能都能够诱导和增殖特异性ctl，但来自患者的特异性ctl的细胞毒性活性比健康志愿者的细胞毒性活性降低。阐明CAEBV患者CTL活性降低的机制，对了解CAEBV的病因和CTL治疗具有重要意义。少

We examined the presence of EB virus (EBV) specific cytotoxic T cells, induction of EBV specific CTLs and the cytotoxic activities of induced CTLs in the peripheral CD8+ cells from the patient with chronic active EBV infection (CAEBV) and the results were as follows,1.Demonstration of EBV specific CTLs by HLAA24-specific and EBV-specific tetramer analysisBRLF1 peptides specific CTLs were found in the peripheral lymphocytes of healthy volunteers and those of patients with infectious mononucleosis. Interestingly, we were also able to demonstrate BRLF1 peptides specific CTLs in the peripheral blood of the patients with CAEBV.2.Induction of antigen specific CTLs :We used HLAA24 positive autologous B lymphobastoid cells (B-LCLs) as antigen presenting cells and peripheral mononuclear cells were stimulated with B-LCLs, pulsed with or without BRLF1 peptide. We were successfully able to induce BRLF1 tetramer positive CTLs even in the absence of BRLF1 specific peptides. This suggested that enoug … More h BRLF1 specific stimulation were given from the EBV infected B-LCLs to specific CTLs without further peptide pulse. These CD8 positive BRLF1 tetramer specific induction and proliferation of CTLs were observed in the mononuclear cells from both healthy volunteers and the CAEBV patient.3.Cytotoxic activity of induced CTLs using interferon gamma producing ability of CTLs :Then we examined the cytotoxic activities of induced CTLs. First we confirmed that BRLF1 tetramer specific CD8+ CTLs from healthy volunteers could produce interferon gamma only after BRLF1 specific peptides. We found that the mononuclear cells from a CAEBV patient also produce BRLF1 tetramer specific CTLs after stimulation with specific peptides. However, BRLF1 specific CTLs from the patient produced interferon gamma only approximately 30%, as compared with those from the healthy volunteers approximately 50%.These results indicated the possibility that mononuclear cells from both healthy volunteers and the CAEBV patients are able to induce and proliferate specific CTLs, but cytotoxic activities of specific CTLs from the patient is reduced as compared with those of healthy volunteers. To elucidate the mechanism of the reduced CTL-activity of the CAEBV patient will have great implications to understand the etiology of CAEBV and the CTL therapy. Less