【作者】 甘露；

【导师】 肖小敏；

【作者基本信息】 暨南大学 ， 妇产科学， 2010， 硕士

【摘要】 目的：探讨乙型肝炎病毒(HBV)感染JAR细胞过程中是否存在抗体依赖病毒增强作用。方法：1.MTT法检测不同浓度的HBIG(终浓度分别为HU/ml,0.1 IU/ml,0.01 IU/ml, 0.001 IU/ml,0.0001 IU/ml)对JAR细胞的作用。2.乙型肝炎病毒血清感染体外培养的JAR细胞,不同浓度的HBIG(终浓度分别为1IU/ml,0.1 IU/ml,0.01 IU/ml,0.001 IU/ml,0.0001 IU/ml)干预,24h后提取细胞内病毒DNA,荧光定量PCR测定细胞内HBV-DNA含量；激光共聚焦检测细胞内HBsAg表达。3.原子力显微镜观察JAR细胞的形态学变化。结果：1.不同浓度的HBIG干预HBV感染的JAR细胞,细胞内HBV-DNA的拷贝数无统计学差异(P>0.05)。2.不同浓度的HBIG干预乙肝病毒感染的JAR细胞,细胞内HBsAg的荧光强度无统计学差异(P>0.05)。3.原子力显微镜观察,阴性对照组的JAR细胞多呈长梭形或多边形；细胞质与细胞核边界清晰,细胞核的体积大,饱满,凸起；细胞向四周延伸出数量不等的伪足样突起；乙型肝炎病毒染JAR细胞后,细胞变为不规则型；细胞核与细胞质分界不清,细胞核不饱满；细胞膜凹陷,细胞膜周围伪足消失。较高浓度的HBIG (0.1IU/ml、0.O1IU/ml)干预乙型肝炎病毒感染的JAR细胞形态与阴性对照组细胞形态相似。一定浓度的HBIG (0.0001 IU/ml) HBIG干预乙型肝炎病毒感染的JAR细胞,细胞变为圆形；细胞核与细胞质分界不清,细胞核不饱满；细胞膜凹陷,细胞膜周围伪足消失。结论：1.高病毒载量的乙型肝炎病毒阳性血清可以感染体外培养的JAR细胞。2.较高浓度的HBIG和最低保护浓度的HBIG不能阻断乙型肝炎病毒感染体外培养的JAR细胞。3.较低浓度的HBIG不能增强乙型肝炎病毒感染体外培养的JAR细胞。4.原子力显微镜观察,HBV感染体外培养的JAR细胞细胞膜存在形态学变化。5.一定浓度的HBIG保护HBV感染的JAR细胞形态接近正常。更多还原

【Abstract】 Objective: To study the antibody-dependent enhancement of HBV infection in the trophoblast cell line JAR cultured in vitro.Method:1. The effect of different concentrations of HBIG(Final concentration, respectively lIU/ml, 0.1IU/ml, 0.01IU/ml, 0.001IU/ml, 0.0001IU/ml) on the JAR cells was analyzed by MTT.2. JAR cells were infected by HBV positive serum of high viral copies and treated with different concentrations of HBIG(Final concentration, respectively 1IU/ml, 0.1IU/ml, 0.01IU/ml,0.001 IU/ml,0.0001 IU/ml). JAR cells were cultivated for 24h, extracted viral DNA from the cells; HBV-DNA was determined by fluorescence quantitative PCR; The expression of the HBsAg in the cells was detected by LSM.3. The morphological changes of JAR were observed cells by AFM.Results:1. There were no differences of HBV-DNA levels in JAR cells with HBV infection treated with different concentrations of HBIG(P＞0.05).2. There were no differences of fluorescence intensity of HBsAg in JAR cells with HBV infection treated with different concentrations of HBIG(P> 0.05).3. By AFM, JAR cells which were not infected by HBV were multi-spindle or polygonal; there was a clear boundary between cytoplasm and nucleus, the size of nucleus was large, full, and protruding; cells extended to every directions, a number of pseudopodia-like protrusions distributed around the cell membrane. JAR cells which were infected by HBV became round or irregular; boundaries between nucleus and cytoplasm was unclear, nucleus was not full; membrane depression, pseudopodia around the cell membrane disappeared. JAR cells with HBV infection treated with different concentrations of HBIG(1IU/ml, O.1IU/ml) were similar with the negative control group. JAR cells with HBV infection treated with HBIG(0.0001IU/ml) were round; boundaries between nucleus and cytoplasm was unclear, nucleus was not full; membrane depression, pseudopodia around the cell membrane disappeared.Conclusions:1. HBV positive serum of high viral copies can infect JAR cells in vitro.2. The groups of high concentration of HBIG and the group of minimum protection concentration of HBIG can not block HBV infect JAR cells in vitro.3. The groups of Low concentration of HBIG can not enhance HBV infect JAR cells in vitro.4. By AFM, there are morphology changes of cell plasma membrane of JAR cells infected by HBV in vitro.5. Some groups of certain concentration of HBIG can protect morphology of JAR cells infected by HBV close to normal.更多还原