Ito Y, Kakumu S, Yoshioka K, Wakita T, Ishikawa T, Koike K
Third Department of Internal Medicine, Nagoya University School of Medicine, Japan.
Gastroenterol Jpn. 1993 Oct;28(5):657-65. doi: 10.1007/BF02806346.
Cytotoxic T lymphocyte (CTL) activity for hepatitis B virus (HBV) DNA-transfected HepG2 cells (designated HB3-5), which secrete HBsAg, HBeAg and HBV particles, was investigated in 31 patients with chronic HBV infection (18 chronic hepatitis and 13 asymptomatic carriers). 51Cr-labeled HB3-5 with A2 as a major HLA class I antigen served as target cells and T cells from peripheral blood mononuclear cells as effector cells. The CTL activity was measured by a 51Cr release assay. Patients were divided into two groups, the A2 group bearing HLA-A2 and the non-A2 group not bearing HLA-A2. Chronic hepatitis patients in the A2 group showed increased HBV Ag-specific cytotoxicity compared with that seen in the non-A2 group (5.2 +/- 3.1% vs. 0.9 +/- 1.4%; means +/- SD, P < 0.01). In the A2 group with chronic hepatitis, the cytotoxicity was greater in anti-HBe positive patients than in HBeAg positive patients (8.6 +/- 1.9% vs. 3.4 +/- 2.0%, P < 0.01), and asymptomatic carriers showed less cytotoxicity (0.35 +/- 0.31%, P < 0.001) compared with chronic hepatitis patients. In the non-A2 group, HBV Ag-specific CTL activity was negligible in most patients and thus no differences were found among all patient groups. The HBV Ag-specific cytotoxicity was inhibited by antibodies to CD3, HLA class I and hepatitis B nucleocapsid antigens. Removal of CD8+ cells also resulted in marked decrease in the cytotoxicity. These findings indicate that HBV Ag-specific cytotoxicity reflects liver cell damage and HBeAg/anti-HBe status. Furthermore, our assay system appears to be useful to assess CTL response in patients with chronic HBV infection.
在31例慢性乙型肝炎病毒（HBV）感染患者（18例慢性肝炎患者和13例无症状携带者）中，研究了针对分泌乙肝表面抗原（HBsAg）、乙肝e抗原（HBeAg）和HBV颗粒的HBV DNA转染的HepG2细胞（命名为HB3-5）的细胞毒性T淋巴细胞（CTL）活性。以A2作为主要HLA I类抗原的51Cr标记的HB3-5细胞作为靶细胞，外周血单个核细胞中的T细胞作为效应细胞。通过51Cr释放试验测量CTL活性。患者分为两组，携带HLA-A2的A2组和不携带HLA-A2的非A2组。与非A2组相比，A2组慢性肝炎患者的HBV抗原特异性细胞毒性增加（5.2±3.1%对0.9±1.4%；平均值±标准差，P<0.01）。在患有慢性肝炎的A2组中，抗HBe阳性患者的细胞毒性大于HBeAg阳性患者（8.6±1.9%对3.4±2.0%，P<0.01），并且无症状携带者的细胞毒性低于慢性肝炎患者（0.35±0.31%，P<0.001）。在非A2组中，大多数患者的HBV抗原特异性CTL活性可忽略不计，因此在所有患者组之间未发现差异。HBV抗原特异性细胞毒性受到抗CD3、HLA I类和乙肝核衣壳抗原抗体的抑制。去除CD8+细胞也导致细胞毒性显著降低。这些发现表明，HBV抗原特异性细胞毒性反映了肝细胞损伤和HBeAg/抗HBe状态。此外，我们的检测系统似乎有助于评估慢性HBV感染患者的CTL反应。
