乙型肝炎病毒基因型B／C型患者血清和肝细胞内HBsAg表达水平的差异  

Affects of HBV genotypes (B/C) on the levels of serum and intrahepatic HBsAg

作　　者：陈金军[1] 马世武[1] 王战会[1] 孙剑[1] 朱幼芙[1] 侯金林[1] 

机构地区：[1]南方医科大学南方医院感染内科,广州510515

出　　处：《中华医学杂志》2006年第28期1947-1951,共5页National Medical Journal of China

基　　金：国家自然科学基金资助项目(G1999054106);国家杰出青年基金资助项目(30225042)

摘　　要：目的了解HBV基因型(B和C)对血清及肝内HBsAg表达的影响。方法选择在广州南方医院感染内科住院的76例慢性乙型肝炎患者,具有同期对应的血清和肝组织切片。用PCR- RFLP方法行HBV基因分型;Abbott Architect法定量检测血清HBsAg;采用单克隆抗体或多克隆抗体对肝脏组织内的HBsAg和HBcAg分别进行免疫组织化学染色,计数阳性细胞数。并对6例基因型B和8例基因型C患者血清HBV HBsAg部分区段DNA进行序列测定。结果共检测到4种基因型:B (47例)、C(21例)、D(3例)和B／C混合(5例)。血清HBsAg水平在基因型B和C之间无区别,但HBV基因C型患者肝内HBsAg的表达高于B型(P<0．01)。在HBV基因B型患者中,HBsAg阳性肝细胞分布多为单个或散在几个(22／47),C型中则多为广泛分布(8／21)(P<0．01)。在基因B型的患者中,血清HBsAg和血清。HBV DNA(r=0．674,P=0．000),核心抗原核内(r=0．534,P=0．000)以及细胞质表达水平(r=0．405,P=0．004)呈正相关关系;在基因C型的患者中,血清HBsAg仅与血清HBV DNA正相关(r=0．503,P=0．017)。肝内HBsAg水平仅在基因B型患者中与肝细胞质内HBcAg的表达水平呈正相关关系(r=0．318,P=0．029),与血清HBsAg、HBV DNA、核心抗原核内表达水平无相关。序列分析显示基因型B和C表面抗原前20个氨基酸序列存在明显差异。结论基因C型患者肝组织中HBsAg表达水平高于B型;而基因B型的血清HBsAg水平和病毒复制水平相关性明显。Objective To observe the effects of HBV genotypes on the level of HBsAg in serum and hepatocytes in chronic hepatitis B patients without antiviral therapy. Methods Seventy-six chronic hepatitis B inpatients were enrolled into this study, and liver biopsies and histologic diagnosis were performed, and serum samples were collected at the time point of liver biopsy. PCR-RFLP method was adopted to determine the genotype of hepatitis B virus and Abbott Architect HBsAg assay was used to quantify the serum HBsAg. Immuostaining for antigens in liver tissues with monoclonal antibody （for HBsAg） or polyclonal antibodies （for HBcAg） was carried out in consecutive slides. The percentages of hepatocytes for HBsAg stain, hepatocytes for HBcAg nuclear stain and hepatocytes for HBcAg cytoplasm stain were estimated in the ranges of 0 （negative）, ≤1%, 1 ＋ ; 2% -5%, 2 ＋ ; 6% -25%, 3 ＋ ; 26% -50%, 4 ＋ ; and 〉50%, 5 ＋. The distributions of positive cells in slides are described as single or isolated, clustery or widespread. Surface gene was directly sequenced with the serum HBV DNA from 6 patients with genotype B and 8 with genotype C HBV infection, respectively. Results Four HBV genotypes were detected in 76 patients： 47 patients with B, 21 with C, 3 with D and 5 were infected by genotype B mixed with C HBV infection. Age, gender, serum HBV DNA level, ALT, AST or histological evaluation （grades and stages scores） were not different between the patients infected with genotype B or C HBV. The level of serum HBsAg was not significantly different between the patients infected with genotype B or C HBV, but the proportions of hepatocytes stained with HBsAg was greater in patients with C type HBV infection than B （P 〈 0. 01 ）. In the liver slides from the patients infected HBV genotype B, HBsAg was stained frequently in single or isolated hepatocytes （22/47）, and widespread HBsAg-positive hepatocytes were often seen in the patients with C type HBV （ 8/21 ）, P 〈 0.01. In the patients with B ty

关 键 词：肝炎病毒 乙型 抗原 表面 基因型 免疫组织化学 

分 类 号：R512.62[医药卫生—内科学]