I. Department of Medicine, Section Infectious Diseases, Hamburg, Germany.
German Center for Infection Research Partner Site, Hamburg, Germany.
J Infect Dis. 2019 Jan 29;219(4):568-577. doi: 10.1093/infdis/jiy549.
This study aimed to comprehensively define the breadth and specificity of the hepatitis delta virus (HDV)-specific T-cell response in patients at different stages of chronic coinfection with hepatitis B virus (HBV).
Following in vitro stimulation with an overlapping set of 21 HDV-specific 20mer peptides and exogenous interleukin 2, HDV-specific CD4+ and CD8+ T-cell responses of 32 HDV-infected patients were analyzed by enzyme-linked immunospot analysis and intracellular cytokine staining for interferon γ production at the single-peptide level. Additionally, HLA-binding studies were performed both in silico and in vitro.
We were able to detect ≥1 T-cell response in >50% our patients. Interestingly, there was no significant difference between the breadth of the response in patients positive and those negative for HDV by PCR. HDV-specific T-cell responses focused on 3 distinct HDV-specific epitopes that were each detected in 12%-21% of patients-2 HLA class II-restricted epitopes (amino acids 11-30 and 41-60) and 1 major histocompatibility complex class I-restricted epitope (amino acids 191-210). In in vitro HLA-binding assays, the 2 CD4+ T-cell specificities (amino acids 11-30 and 41-60) showed promiscuous binding to multiple HLA-DR molecules.
This comprehensive characterization of HDV T-cell epitopes provides important information that will facilitate further studies of HDV immunopathogenesis.
本研究旨在全面定义慢性乙型肝炎病毒(HBV)合并感染患者不同阶段肝δ病毒(HDV)特异性 T 细胞反应的广度和特异性。
通过用重叠的 21 个 HDV 特异性 20 肽和外源性白细胞介素 2 体外刺激,通过酶联免疫斑点分析和细胞内细胞因子染色分析干扰素 γ产生,对 32 例 HDV 感染患者的 HDV 特异性 CD4+和 CD8+ T 细胞反应进行分析,以检测单个肽水平。此外,还进行了 HLA 结合的计算机模拟和体外研究。
我们能够在>50%的患者中检测到≥1 个 T 细胞反应。有趣的是,PCR 阳性和阴性的患者反应广度没有显著差异。HDV 特异性 T 细胞反应集中在 3 个不同的 HDV 特异性表位上,每个表位在 12%-21%的患者中被检测到,包括 2 个 HLA Ⅱ类限制性表位(氨基酸 11-30 和 41-60)和 1 个主要组织相容性复合体Ⅰ类限制性表位(氨基酸 191-210)。在体外 HLA 结合实验中,2 个 CD4+ T 细胞特异性(氨基酸 11-30 和 41-60)显示出与多种 HLA-DR 分子的混杂结合。
对 HDV T 细胞表位的全面表征提供了重要信息,将有助于进一步研究 HDV 免疫发病机制。