我国HCV主要基因型优势抗原表位的筛选

发布时间：2018-03-28 01:29

本文选题：丙型肝炎病毒　切入点：多价疫苗　出处：《实用检验医师杂志》2015年04期

【摘要】：目的针对我国丙型肝炎病毒(hepatitis C virus,HCV)主要基因型筛选出能有效诱导体液免疫和细胞免疫的抗原多肽。方法利用生物信息学方法预测评估我国HCV主要基因型1b、2a、6a型的优势B、CTL和Th抗原表位。以HCV-CTL、HCV-B+HCV-Th、HCV-CTL+HCV-B+HCV-Th组合肽作为免疫原免疫BALB/C小鼠,经乳酸脱氢酶释放法检测CTL杀伤活性、ELISPOT法检测分泌IFN-γ能力、ELISA法检测抗体滴度,对检测结果进行统计学分析。结果 SP2/0对照细胞未产生特异性免疫应答,而SP2/0-NS3实验细胞HCV-CTL+HCV-B+HCV-Th和HCV-CTL均诱导了特异性的CTL应答,且当效靶比为25∶1和50∶1时,三组间差异均有统计学意义(F=2045,F=6338,P均0.05),且HCVCTL+HCV-B+HCV-Th联合免疫激活的CTL活性高于HCV-CTL单独免疫,HCV-CTL单独免疫高于PBS空白对照,差异均有统计学意义(P均0.05)。细胞体液联合免疫、单独体液免疫、单独细胞免疫刺激的分泌IFN-γ的细胞数量差异有统计学意义(F=5308,P0.05)。单独体液免疫、单独细胞免疫刺激的分泌IFN-γ的细胞数量均低于细胞、体液联合免疫,单独体液免疫刺激的分泌IFN-γ的细胞数量高于单独细胞免疫,且差异均有统计学意义(P均0.05)。不同时间点不同免疫原所诱导的特异性的Ig G抗体滴度差异均有统计学意义(P均0.01)。不同时间点用HCV-CTL+HCV-B+HCV-Th联合免疫和HCV-B+HCV-Th免疫所诱导的特异性Ig G抗体滴度均显著高于PBS空白对照,且差异均有统计学意义(P均0.05),而细胞体液联合免疫与单独体液免疫之间特异性Ig G抗体滴度经比较,差异无统计学意义(P0.05)。结论 HCV-CTL+HCV-B+HCV-Th联合免疫诱导了高效的体液免疫和细胞免疫应答,有望进一步开发为有效的重组HCV疫苗。
[Abstract]:Objective to screen out the antigenic polypeptides of hepatitis C virus hepatitis C virus (HCV) genotype which can effectively induce humoral and cellular immunity in China. Methods the bioinformatics method was used to predict and evaluate the superiority of the main genotype of HCV 1bm2ah6a in China. HCV-CTL and Th epitopes were immunized with HCV-CTL hHCV-B HCV-Th-HCV-CTL HCV-B HCV-Th combined peptide as immunogen to immunize BALB/C mice. The cytotoxic activity of CTL was detected by lactate dehydrogenase release assay and Elisa was used to detect the titer of antibody secreting IFN- 纬. The results were statistically analyzed. Results the SP2/0 control cells did not produce a specific immune response. Both HCV-CTL HCV-B HCV-Th and HCV-CTL induced specific CTL responses, and the specific CTL responses were observed at 25:1 and 50:1, respectively. The difference among the three groups was statistically significant (P < 0.05), and the activity of CTL activated by HCVCTL HCV-B HCV-Th was higher than that by HCV-CTL alone, and the difference was statistically significant (P < 0.05). The number of IFN- 纬 secreted by single cellular immunostimulation was significantly different. The number of IFN- 纬 secreting cells stimulated by single humoral immunity was lower than that of cells, and the number of IFN- 纬 secreted by single humoral immunity was lower than that by humoral combined immunization. The number of IFN- 纬 secreting cells stimulated by humoral immunity alone was higher than that of single cellular immunity. The titers of specific IgG antibody induced by different immunogen at different time points were significantly different (P < 0.01). HCV-CTL HCV-B HCV-Th combined immunization and HCV-B HCV-Th immunization at different time points were used to induce Ig G antibody titers. The titers of specific IgG antibody were significantly higher than those of PBS control. The difference was statistically significant (P < 0.05), and the titers of specific IgG antibody between cellular humoral combined immunity and humoral immunity alone were compared. Conclusion combined immunization with HCV-CTL HCV-B HCV-Th can induce highly effective humoral and cellular immune responses, which is expected to be an effective recombinant HCV vaccine.
【作者单位】：江西省肿瘤医院检验科;江西省儿童医院检验科;
【基金】：江西省科技厅项目(20122BAB215033)
【分类号】：R373.21

【共引文献】