摘要
目的 :探讨乙型肝炎病毒 (HBV) pre S2 - S基因诱发的体液免疫在乙型肝炎发病机制中的作用。 方法 :用含 HBVpre S2 - S基因的质粒 pc DNA3- S2 - S经胫骨前肌肌肉注射免疫正常 BAL B/ c小鼠获得抗血清 ,将抗血清经尾静脉注射到转基因小鼠 BAL B/ c- Tg N(pre S2 - S/ ayw)体内 ,不同时间点静脉采血检测小鼠血清 HBs Ag、抗 HBs抗体、前 S2抗原、前 S2抗体、转氨酶、尿素氮和肌酐的变化 ,并处死动物检查肝、脾、肾、肠、肺和肌肉组织的病理学改变。 结果 :DNA免疫正常小鼠后 8～14周 ,抗 HBs抗体的浓度维持在 130～ 14 0 m IU/ ml;免疫后小鼠抗血清转移到转基因小鼠 BAL B/ c- Tg N(pre S2 - S/ ayw)后 2、4、7和 14 d,小鼠血清中未检测到 HBs Ag和前 S2抗原 ,抗 HBs抗体持续阳性 ,前 S2抗体 14 d时转阴 ;AL T2、4 d升高 ,7d恢复到正常 ;AST14 d内始终高于正常值 ,4 d时达高峰 ;尿素氮异常 ,γ- GT与肌酐正常。转基因小鼠肝脏出现急性乙型肝炎的改变 ,初期 (2 d、4 d)血窦内和汇管区有淋巴细胞浸润 ,4 d肝细胞开始肿胀 ;中期 (7d)全小叶出现肝细胞气球样变 ,肝小叶内出现肝细胞点灶性坏死伴单个核细胞浸润 ,汇管区轻度单个核细胞的浸润 ;后期 (14 d)时肝细胞肿胀明显减轻 。
Objective:To study the liver lesion resulted from humoral immune response induced by hepatitis B virus preS2 and S genes in hepatitis B transgenic mice BALB/c TgN( preS2 S /ayw). Methods:Anti sera were obtained from the normal BALB/c mice injected with 100 μg recombinant DNA (pcDNA3 S2 S) encoding HBV middle envelope protein. An HBV hepatitis B transgenic mouse lineage BALB/c TgN( preS2 S /ayw) expressing noncytotoxic transgenic mouse model system was developed to analyze the immunopathogenesis of HBV induced liver disease. Anti sera were collected and transfered into transgenic mice(habouring HBV preS2 and S genes) and their nontransgenic littermates. Blood was collected from anesthetized mice by retrobulbar puncture using heparinized glass pipettes at various intervals after immunization to measure the level of HBsAg anti HBs antibody preS2 antigen anti preS2 antibody ALT,AST,Cr and Urea.Mice were sacrificed at various intervals after transfered.Tissues were fixed in alcoholic Bouins fixative.Sections of paraffin embedded tissues were cut and stained with hematoxylin/eosin. Results:The concentration of anti HBs antibody was 130 140 mIU/ml 8 14 weeks after immunization.HBsAg and preS2 antigen remained undetectable after passive transfer of anti sera.Anti HBs antibody could be detectable till 14th day.Anti preS2 antibody could not be detected by the 14th day. The serum level of transaminases (ALT) were elevated on the 2nd d and the 4th d. The serum level of urea were higher than normal.The liver of the transgene mice resembled the acute hepatitis B.The early phase of injury(2nd d and 4th d) was a few lymphomononuclear cells infiltrate on portal areas and sinusoid.Hepatocytes began to degenerate on 4th day. The middle phase of injury(7th d) had widespread ballooning degeneration of hepatocytes and focal hepatocellular necrosis,complicated with parenchymal lymphoid infiltration and lymphomononuclear cells infiltration in portal areas. The late phase of injury(14th d) showed that swelling of
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2003年第2期159-163,共5页
Academic Journal of Second Military Medical University
基金
国家"九五"攻关项目 (TJ99-LA0 1)
国家自然科学基金 (3 9670 811)
上海市科学技术发展基金项目 (9949190 3 3 )
关键词
乙型肝炎病毒
preS2-S基因
体液免疫
转基因小鼠
乙型肝炎
hepatitis B virus
preS2 S gene
humoral immunization
transgenic mice
hepatitis B [Acad J Sec Mil Med Univ,2003,24(2):159 163]
