HBV阳性血清体外感染人绒毛膜滋养细胞株JEG-3的研究

Study on the effect of positive HBV serum in infecting human chorionic trophocyte in vitro

目的 建立乙型肝炎病毒（hepatitis B virus,HBV）体外感染人绒毛膜滋养细胞株JEG-3的模型,探讨HBV宫内感染的可能危险因素.方法 对人绒毛膜滋养细胞株JEG-3进行传代培养,将不同复制水平的HBV阳性血清与传代培养的JEG-3细胞在无人为干扰因素下共同孵育48~72小时,实时荧光定量PCR方法检测细胞内HBV-DNA的复制水平.结果 在无人为干扰因素下,HBV阳性血清体外可感染人绒毛膜滋养细胞株JEG-3,正常人血清细胞中未能检测出HBV-DNA;低、中危组细胞内也未检测出HBV-DNA;高危组复制水平为4.56×103U/mL,极高危组复制水平为2.72×104U/mL,随着HBV阳性血清复制水平的提高,JEG-3细胞内HBV复制水平有升高的趋势.结论 HBV体外可感染人绒毛膜滋养细胞株JEG-3,HBV高复制水平可能是其宫内感染的独立危险因素之一.

Objective To establish the model of Hepatitis B virus （HBV） infecting human chorionic trophocyte line JEG-3 in vitro so as to discusses the possible risk factors of HBV intrauterine infection. Methods Human chorionic trophocyte line JEG-3 in early pregnancy was subcuhured, then the HBV serum with different replication levels and subcuhured cells JEG-3 were incubated together for 48-72hrs without any artificial interference factors. Real-time fluorescence quantitative PCR （RT-PCR） was employed to detect the replication level of intracellular HBV-DNA. Results Without artificial interference factors HBV positive serum could infect the human chorionic trophocyte line JEG-3 in vitro, and HBV-DNA failed to be detected in normal human serum. HBV-DNA also failed to be detected in low risk and medium risk group. HBV replication level of high risk group was 4.56 × 103U/mL, and that of highest risk group was 2.72 × 104U/mL. With the rising of replication level of positive HBV serum, HBV replication level in JEG-3 cells increased. Conclusion HBV can infect human chorionic trophocyte line JEG-3 in vitro, and high replication level of HBV may be one of the independent risk factors of intrauterine infection.