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目的比较不同质粒建立的水动力转染乙型肝炎小鼠模型的特性。方法将pc DNA 3.1-1.3-HBV-C与PAAV-1.2-HBV-A两种质粒通过水动力转染法建立两种乙型肝炎小鼠模型,通过对建模成功率和小鼠外周血HBs Ag滴度及稳定性,T细胞指数流式等检测分析,比较不同模型差异和特效。结果转染pc DNA3.1-1.3-HBV-C质粒建模成功率为78.6%,转染后体内HBs Ag滴度最高达到220IU/m L;转染PAAV-1.2-HBV-A质粒建模成功率为56%,转染后体内HBs Ag滴度最高达到2 300IU/m L。建模小鼠的肝脏进行切片,做免疫组化后,观察实验结果,转染pc DNA 3.1-1.3-HBV-C质粒的小鼠肝脏切片的HBs Ag免疫组化较PAAV-1.2-HBV-A质粒小鼠颜色面积更大。实验小鼠转染质粒后6周的外周血进行流式细胞术检测,结果显示特异性T细胞CD4+、CD8+和正常小鼠比较,两种质粒的造模小鼠未有差别。结论利用水动力转染法用不同质粒建造乙型肝炎小鼠模型在乙肝病毒指标水平和稳定性方面有着显著差异。
OBJECTIVE: To compare the characteristics of different types of plasmids in establishing a model of hydro-transfected hepatitis B mice. Methods Two hepatitis B mouse models were established by hydrodynamic transfection of pcDNA3.1-1.3-HBV-C and PAAV-1.2-HBV-A plasmids. The success rate of modeling and HBsAg Ag titer and stability, T cell index flow analysis and other tests to compare different model differences and special effects. Results The successful rate of pcDNA3.1-1.3-HBV-C plasmid transfection was 78.6%, and the highest HBsAg titer was 220IU / ml after transfection. The plasmid transfected with PAAV-1.2-HBV-A was successfully constructed The rate was 56%. The highest HBsAg titer reached 2,300 IU / mL after transfection. The liver of the model mice was sectioned and immunohistochemically. After the results of the experiment were observed, the HBs Ag immunohistochemistry of the mouse liver sections transfected with the pcDNA 3.1-1.3-HBV-C plasmid was more effective than that of the PAAV-1.2-HBV-A Plasmids mice have a larger color area. Flow cytometry was used to detect the peripheral blood of experimental mice after transfection of plasmid for 6 weeks. The results showed that compared with normal mice, CD4 + and CD8 + of specific T cells showed no difference between the two plasmids. Conclusion The construction of Hepatitis B mouse model by using different plasmids using hydrodynamic transfection method has significant difference in the level and stability of hepatitis B virus.