The cytotoxic T lymphocyte response to multiple hepatitis B virus polymerase epitopes during and after acute viral hepatitis.
The cytotoxic T lymphocyte response to multiple hepatitis B virus polymerase epitopes during and after acute viral hepatitis.
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DOI：
10.1084/jem.181.3.1047
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发表时间：
1995-03-01
期刊：
影响因子：
--
通讯作者：
Chisari FV
中科院分区：
文献类型：
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作者：
Rehermann B;Fowler P;Sidney J;Person J;Redeker A;Brown M;Moss B;Sette A;Chisari FV
细胞毒性T淋巴细胞（CTL）被认为在B型肝炎病毒（HBV）感染过程中有助于病毒清除和肝细胞损伤。使用的策略，涉及在体外刺激外周血单核细胞（PBMC）与HBV衍生的合成肽含有HLA-A2.1，-A31，和-Aw 68结合基序，我们以前描述了CTL反应的几个表位内的HBV核衣壳和包膜抗原在急性肝炎患者。在这项研究中，我们定义了六个HLA-A2限制性CTL表位位于高度保守的逆转录酶和RNA酶H结构域的病毒聚合酶蛋白，我们表明，聚合酶的CTL反应是多克隆的，多特异性的，并介导的CD 8 + T细胞在急性病毒性肝炎患者，但它是检测不到的慢性HBV感染患者或未感染的健康献血员。重要的是，肽激活的CTL识别表达内源性合成的聚合酶蛋白的靶细胞，表明这些肽代表天然加工的病毒表位。对肽刺激无反应的患者的病毒的DNA序列分析表明，CTL无反应性不是由于与合成肽序列不同的病毒变体感染所致。对其中一个表位具有特异性的CTL不能识别几种天然存在的病毒变体，除非在高肽浓度下，这强调了这种应答的HBV亚型特异性。此外，针对聚合酶，核心和包膜表位的CTL应答在完全临床恢复和血清转换后一年多内可检测到，反映了在没有病毒抗原的情况下痕量病毒的持续存在或长寿命记忆CTL的存在。最后，我们证明，野生型病毒DNA和RNA可以无限期地持续存在，微量，在血清和PBMC后，完全临床和血清学恢复，尽管伴随的，有力的，和持续的多克隆CTL反应。由于病毒的持久性不是由于逃避CTL识别在这些条件下，数据表明，HBV可能会撤退到免疫豁免网站，它可以种子循环和到达CTL无法访问的组织，从而保持CTL反应在明显治愈的个人，也许，延长慢性肝炎患者的肝脏疾病。
Cytotoxic T lymphocytes (CTL) are thought to contribute to viral clearance and liver cell injury during hepatitis B virus (HBV) infection. Using a strategy involving the in vitro stimulation of peripheral blood mononuclear cells (PBMC) with HBV-derived synthetic peptides containing HLA-A2.1, -A31, and -Aw68 binding motifs, we have previously described CTL responses to several epitopes within the HBV nucleocapsid and envelope antigens in patients with acute hepatitis. In this study we define six HLA-A2-restricted CTL epitopes located in the highly conserved reverse transcriptase and RNase H domains of the viral polymerase protein, and we show that the CTL response to polymerase is polyclonal, multispecific, and mediated by CD8+ T cells in patients with acute viral hepatitis, but that it is not detectable in patients with chronic HBV infection or uninfected healthy blood donors. Importantly, the peptide-activated CTL recognize target cells that express endogenously synthesized polymerase protein, suggesting that these peptides represent naturally processed viral epitopes. DNA sequence analysis of the viruses in patients who did not respond to peptide stimulation indicated that CTL nonresponsiveness was not due to infection by viral variants that differed in sequences from the synthetic peptides. CTL specific for one of the epitopes were unable to recognize several naturally occurring viral variants, except at high peptide concentration, underlining the HBV subtype specificity of this response. Furthermore, CTL responses against polymerase, core, and envelope epitopes were detectable for more than a year after complete clinical recovery and seroconversion, reflecting either the persistence of trace amounts of virus or the presence of long lived memory CTL in the absence of viral antigen. Finally, we demonstrated that wild type viral DNA and RNA can persist indefinitely, in trace quantities, in the serum and PBMC after complete clinical and serological recovery, despite a concomitant, vigorous, and sustained polyclonal CTL response. Since viral persistence is not due to escape from CTL recognition under these conditions, the data suggest that HBV may retreat into immunologically privileged sites from which it can seed the circulation and reach CTL-inaccessible tissues, thereby maintaining the CTL response in apparently cured individuals and, perhaps, prolonging the liver disease in patients with chronic hepatitis.