Pasquinelli C, Melegari M, Villa E, Scaglioni P P, Seidenari M, Mongiardo N, De Rienzo B, Manenti F
Chair of Gastroenterology, University of Modena, Italy.
J Med Virol. 1990 Jun;31(2):135-40. doi: 10.1002/jmv.1890310211.
Hepatitis B virus (HBV) infection of peripheral blood mononuclear cells (PBMCs) has been observed in all stages of liver disease. Thus far all information about the physical state of HBV in mononuclear blood cells comes from Southern blot analysis and in situ hybridization. In this study we focused our attention on the presence of HBV DNA sequences in PBMCs of 30 patients with acute type B hepatitis and 6 patients with chronic active hepatitis by utilizing both Southern blot analysis and the polymerase chain reaction (PCR). Southern blot analysis showed no HBV DNA sequences in PBMCs of the acute hepatitis patients, although the sensitivity of our method enabled us to detect as little as 1 pg of cloned HBV insert. As far as the chronic hepatitis patients are concerned Southern blot analysis revealed the presence of HBV DNA sequences in 5 out of 6 patients but intermittently at successive follow-up times. On the other hand we were able to demonstrate the presence of HBV related sequences in 14 out of 30 acute hepatitis patients (5 HBeAg positive, 9 antiHBe positive) and in all 6 chronic hepatitis patients by PCR. Our results indicate that the involvement of PBMCs with HBV during acute HBV infection occurs at a very low level, often below the detection limit of the Southern blot technique.
在肝病的各个阶段均观察到外周血单个核细胞(PBMC)感染乙型肝炎病毒(HBV)。迄今为止,关于单核血细胞中HBV物理状态的所有信息均来自Southern印迹分析和原位杂交。在本研究中,我们通过Southern印迹分析和聚合酶链反应(PCR),重点关注了30例急性乙型肝炎患者和6例慢性活动性肝炎患者PBMC中HBV DNA序列的存在情况。Southern印迹分析显示,急性肝炎患者的PBMC中未检测到HBV DNA序列,尽管我们的方法灵敏度能够检测低至1 pg的克隆HBV插入片段。就慢性肝炎患者而言,Southern印迹分析显示6例患者中有5例存在HBV DNA序列,但在连续随访期间呈间歇性出现。另一方面,通过PCR我们能够证明30例急性肝炎患者中有14例(5例HBeAg阳性,9例抗-HBe阳性)以及所有6例慢性肝炎患者中存在HBV相关序列。我们的结果表明,急性HBV感染期间PBMC与HBV的关联发生水平非常低,通常低于Southern印迹技术的检测限。