Vassilopoulos D, Rapti I, Nikolaou M, Hadziyannis E, Hadziyannis S J
Academic Department of Medicine, Athens University School of Medicine, Hippokration General Hospital, Athens, Greece.
J Viral Hepat. 2008 Nov;15(11):817-26. doi: 10.1111/j.1365-2893.2008.00996.x. Epub 2008 Jun 11.
The immunopathogenesis of hepatitis B e antigen (HBeAg) negative chronic hepatitis B (CHB) virus (HBV) infection has not been adequately investigated. We studied the cellular immune responses of peripheral lymphocytes using proliferating assays, intracellular cytokine staining (ICS) and ELISPOT interferon-gamma (IFN-gamma) assays after non-specific and specific stimulation with whole HBV proteins and synthetic peptides. Thirty patients with HBeAg negative CHB, eleven HBsAg inactive carriers, nine patients with acute hepatitis B and 22 healthy controls were included in the study. Patients with HBeAg negative CHB demonstrated an increased number of peripheral CD8+ T cells while their peripheral blood mononuclear cells showed increased proliferation after in vitro stimulation with overlapping hepatitis B core derived peptides and an envelope derived epitope (HBs 182-191 aa), similar to those observed in acute hepatitis B. Using ICS, we found an expanded population of IFN-gamma producing T lymphocytes, CD4+ and CD8+, after non-specific stimulation, in HBeAg negative CHB compared to all other groups. HBeAg negative CHB and acute hepatitis B patients had a similarly increased number of core specific T cells measured by the IFN-gamma assays. Inactive HBsAg carriers showed minimal proliferative responses overall while they exhibited an increased number of envelope specific effector T cells (measured by ICS). In conclusion, we showed that overall CD4+ T cell responses from patients with HBeAg negative CHB were comparable to those of acute hepatitis B, while inactive HBsAg carriers despite their limited proliferative capacity the effector activity of their peripheral T cells was maintained.
乙肝e抗原（HBeAg）阴性慢性乙型肝炎（CHB）病毒（HBV）感染的免疫发病机制尚未得到充分研究。我们使用增殖试验、细胞内细胞因子染色（ICS）和酶联免疫斑点法检测干扰素-γ（IFN-γ），研究了外周淋巴细胞在用全HBV蛋白和合成肽进行非特异性和特异性刺激后的细胞免疫反应。本研究纳入了30例HBeAg阴性CHB患者、11例HBsAg非活动性携带者、9例急性乙型肝炎患者和22名健康对照。HBeAg阴性CHB患者外周血CD8+T细胞数量增加，而其外周血单个核细胞在用重叠的乙肝核心衍生肽和包膜衍生表位（HBs 182-191氨基酸）进行体外刺激后增殖增加，这与急性乙型肝炎患者中观察到的情况相似。通过ICS，我们发现与所有其他组相比，HBeAg阴性CHB患者在非特异性刺激后，产生IFN-γ的T淋巴细胞（CD4+和CD8+）群体有所扩大。通过IFN-γ检测，HBeAg阴性CHB患者和急性乙型肝炎患者的核心特异性T细胞数量同样增加。HBsAg非活动性携带者总体上表现出最小的增殖反应，而通过ICS检测，他们的包膜特异性效应T细胞数量增加。总之，我们发现HBeAg阴性CHB患者的总体CD4+T细胞反应与急性乙型肝炎患者相当，而HBsAg非活动性携带者尽管增殖能力有限，但其外周T细胞的效应活性得以维持。
