Piriou Erwan R, van Dort Karel, Nanlohy Nening M, van Oers Marinus H, Miedema Frank, van Baarle Debbie
Department of Clinical Viro-Immunology, Sanquin Research at CLB and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Eur J Immunol. 2005 Mar;35(3):796-805. doi: 10.1002/eji.200425792.
A lower function of EBV-specific CD8(+) T cells in HIV-infected subjects could be related to a lack of specific CD4(+) T cell help. Therefore, we studied EBV-specific CD4(+) T cells in both healthy donors and untreated or highly active antiretroviral therapy (HAART)-treated HIV-seropositive homosexual men. To this end, PBMC were stimulated with overlapping peptide pools from a latent and a lytic EBV protein, EBV nuclear antigen (EBNA)1 and EBV lytic-switch protein ZEBRA (BZLF1), respectively. EBV-specific CD4(+) T cell frequencies measured directly ex vivo were low. To measure EBV-specific memory CD4(+) T cells, capable of both expansion and IFN-gamma production upon antigenic challenge, we developed a specific and reproducible assay, combining ex vivo expansion of specific T cells with flow cytometric analysis of IFN-gamma production. Untreated HIV-infected individuals had a lower CD4(+) T cell response to both EBNA1 and BZLF1 as compared to healthy EBV carriers and HAART-treated HIV-positive subjects. This suggests loss of EBV-specific CD4(+) T cells due to HIV infection, while HAART might restore this response. In addition, we found an increase in the EBNA1-specific CD8(+) T cell response in HAART-treated subjects. Interestingly, numbers of EBV-specific CD4(+) and CD8(+) T cells were inversely correlated with EBV viral load, suggesting an important role also for EBV-specific CD4(+) T cells in the control of EBV infection.
在HIV感染的受试者中，EBV特异性CD8(+) T细胞功能降低可能与缺乏特异性CD4(+) T细胞辅助有关。因此，我们研究了健康供者以及未经治疗或接受高效抗逆转录病毒治疗（HAART）的HIV血清阳性同性恋男性中的EBV特异性CD4(+) T细胞。为此，分别用来自潜伏性和裂解性EBV蛋白（EBV核抗原（EBNA）1和EBV裂解开关蛋白ZEBRA（BZLF1））的重叠肽池刺激外周血单核细胞（PBMC）。直接在体外测量的EBV特异性CD4(+) T细胞频率较低。为了测量能够在抗原刺激后进行扩增和产生γ干扰素的EBV特异性记忆CD4(+) T细胞，我们开发了一种特异性且可重复的检测方法，将特异性T细胞的体外扩增与γ干扰素产生的流式细胞术分析相结合。与健康的EBV携带者和接受HAART治疗的HIV阳性受试者相比，未经治疗的HIV感染个体对EBNA1和BZLF1的CD4(+) T细胞反应均较低。这表明HIV感染导致EBV特异性CD4(+) T细胞丧失，而HAART可能恢复这种反应。此外，我们发现接受HAART治疗的受试者中EBNA1特异性CD8(+) T细胞反应增加。有趣的是，EBV特异性CD4(+)和CD(+) T细胞数量与EBV病毒载量呈负相关，这表明EBV特异性CD4(+) T细胞在控制EBV感染中也起重要作用。
