Braendstrup Peter, Mortensen Bo Kok, Justesen Sune, Osterby Thomas, Rasmussen Michael, Hansen Andreas Martin, Christiansen Claus Bohn, Hansen Morten Bagge, Nielsen Morten, Vindeløv Lars, Buus Søren, Stryhn Anette
Laboratory of Experimental Immunology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark; The Allogeneic Hematopoietic Cell Transplantation Laboratory, Department of Hematology, Rigshospitalet, Copenhagen University Hospital, Copenhagen, Denmark.
Laboratory of Experimental Immunology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark.
PLoS One. 2014 Apr 23;9(4):e94892. doi: 10.1371/journal.pone.0094892. eCollection 2014.
Human cytomegalovirus (HCMV) is an important human pathogen. It is a leading cause of congenital infection and a leading infectious threat to recipients of solid organ transplants as well as of allogeneic hematopoietic cell transplants. Moreover, it has recently been suggested that HCMV may promote tumor development. Both CD4+ and CD8+ T cell responses are important for long-term control of the virus, and adoptive transfer of HCMV-specific T cells has led to protection from reactivation and HCMV disease. Identification of HCMV-specific T cell epitopes has primarily focused on CD8+ T cell responses against the pp65 phosphoprotein. In this study, we have focused on CD4+ and CD8+ T cell responses against the immediate early 1 and 2 proteins (IE1 and IE2). Using overlapping peptides spanning the entire IE1 and IE2 sequences, peripheral blood mononuclear cells from 16 healthy, HLA-typed, donors were screened by ex vivo IFN-γ ELISpot and in vitro intracellular cytokine secretion assays. The specificities of CD4+ and CD8+ T cell responses were identified and validated by HLA class II and I tetramers, respectively. Eighty-one CD4+ and 44 CD8+ T cell responses were identified representing at least seven different CD4 epitopes and 14 CD8 epitopes restricted by seven and 11 different HLA class II and I molecules, respectively, in total covering 91 and 98% of the Caucasian population, respectively. Presented in the context of several different HLA class II molecules, two epitope areas in IE1 and IE2 were recognized in about half of the analyzed donors. These data may be used to design a versatile anti-HCMV vaccine and/or immunotherapy strategy.
人巨细胞病毒（HCMV）是一种重要的人类病原体。它是先天性感染的主要原因，也是实体器官移植受者以及异基因造血细胞移植受者面临的主要感染威胁。此外，最近有研究表明HCMV可能促进肿瘤发展。CD4+和CD8+ T细胞反应对于病毒的长期控制都很重要，过继转移HCMV特异性T细胞可预防病毒再激活和HCMV疾病。HCMV特异性T细胞表位的鉴定主要集中在针对pp65磷蛋白的CD8+ T细胞反应上。在本研究中，我们聚焦于针对即刻早期1和2蛋白（IE1和IE2）的CD4+和CD8+ T细胞反应。使用覆盖整个IE1和IE2序列的重叠肽，通过体外IFN-γ ELISpot和体外细胞内细胞因子分泌试验对16名健康的、进行了HLA分型的供者的外周血单个核细胞进行筛选。CD4+和CD8+ T细胞反应的特异性分别通过HLA II类和I类四聚体进行鉴定和验证。共鉴定出81个CD4+和44个CD8+ T细胞反应，分别代表至少7个不同的CD4表位和14个CD8表位，这些表位分别受7种和11种不同的HLA II类和I类分子限制，总共分别覆盖了91%和98%的高加索人群。在几种不同的HLA II类分子背景下，约一半的分析供者识别出了IE1和IE2中的两个表位区域。这些数据可用于设计通用的抗HCMV疫苗和/或免疫治疗策略。
