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

我们检测了慢性活动性EBV感染（CAEBV）患者外周血CD 8+细胞中EB病毒（EBV）特异性细胞毒性T细胞的存在、EBV特异性CTL的诱导以及诱导的CTL的细胞毒性活性，结果如下：1.通过HLAA 24特异性和EBV-CTL证实EBV特异性CTL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BRLF 1肽特异性CTL。2.抗原特异性CTL的诱导：以HLA A 24阳性的自体B类淋巴母细胞（B-LCL）为抗原提呈细胞，用B-LCL刺激外周血单个核细胞，用BRLF 1肽或不用BRLF 1肽刺激。即使在没有BRLF 1特异性肽的情况下，我们也能够成功地诱导BRLF 1四聚体阳性CTL。这表明， 关于我们 h BRLF 1特异性刺激从EBV感染的B-LCL给予特异性CTL而无需进一步肽脉冲。在健康志愿者和CAEBV患者的单个核细胞中观察到这些CD 8阳性BRLF 1四聚体特异性诱导和增殖的CTL。3.利用CTL产生干扰素γ的能力检测诱导的CTL的细胞毒活性：然后检测诱导的CTL的细胞毒活性。首先，我们证实了来自健康志愿者的BRLF 1四聚体特异性CD 8 + CTL只能在BRLF 1特异性肽之后产生干扰素γ。我们发现来自CAEBV患者的单核细胞在用特异性肽刺激后也产生BRLF 1四聚体特异性CTL。结果表明，正常人和CAEBV患者外周血单个核细胞均能诱导增殖特异性CTL，但患者外周血特异性CTL的细胞毒活性较正常人低。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