免疫组化方法观察乙肝病毒感染肝星状细胞的研究

Immunohistochemistry study on the infectivity of hepatitis B viruses on human hepatic stellate cells

目的研究HBV能否体外感染LX-2细胞，以期为阐明慢性乙肝的致病机制提供新的实验依据。方法体外培养LX-2细胞细胞数目达到10^6培养瓶时，用HBV感染者的血清进行感染，HBVDNA终浓度分别为10^4、10^5、10^6、10^7拷贝／mL，同一浓度的感染时间均为24、48和72h。按照观察时间点收取细胞，免疫组织化学法（DAB显色）测定HBV在细胞内是否表达乙肝表面抗原（HBsAg）和乙肝核心抗原（HBcAg）。结果用免疫组化方法标记HBsAg和HBcAg时，LX-2细胞中未发现阳性颗粒；HepG2．2．15细胞中有少量棕黄色颗粒，位于胞质中；慢性乙肝患者肝组织中可见大量棕黄色颗粒。结论在体外培养中HBV不能感染LX-2细胞和进行抗原表达。

The purpose of this study was to observe whether HBV can infect HSC in vitro. LX-2 cells, the human activated HSC cell line, were incubated with human serum containing HBVDNA at final concentrations from 0.01 to 10 copies/cell and harvested after 24,48 and 72 hours. Hepatitis B surface antigen （HBsAg） and hepatitis B core antigen（HBcAg） were detected by Immunohistochemistry dyed by DAB. By Immunohistochemistry, no positive particles in LX-2 cells, a few of dark brown particles （ HBsAg and HB- cAg） were found in cytoplasm and nucleoli of HepG2.2.15 cells ,a lot of dark brown particles were found in liver biopsies of patients with chronic hepatitis B. In vitro, there was no evidence that HBV can infect and express antigens in LX-2 cells.