布鲁氏菌Omp2b优势T-B联合抗原表位的免疫应答特点研究

目的 通过生物信息技术预测分析Omp2b蛋白的优势B细胞表位和T细胞表位,筛选T-B联合抗原表位,探讨其免疫原性和激发免疫应答的特点,为开发有效的布鲁氏菌疫苗奠定基础。方法 1)利用生物信息学软件ProtParam,SOMPA,SWISS-MODEL,Rasmol,DNAStar,SYFPEITHI和IEDB来分析Omp2b蛋白的结构,预测T细胞和B细胞的优势表位以及T-B联合表位;2)ELISPOT法检测细胞中IFN-γ阳性细胞数;3)ELISA检测布鲁氏菌病患者血清中针对Omp2b蛋白的T-B联合肽的特异性IgG抗体水平。体外培养免疫细胞,检测上清液中的穿孔素和颗粒酶B。结果 生物信息软件综合分析Omp2b蛋白并筛选出了1个T-B细胞联合抗原表位的潜在区段196-216;患者组中分泌IFN-γ细胞数(SFU)较健康对照组增高(F=25.413,P<0.01),布鲁氏菌病患者血清中的IgG抗体、穿孔素和颗粒酶B的浓度也增加(F=13.653,P<0.01)。结论 Omp2b蛋白的T-B联合抗原表位可以产生特异性的体液免疫应答和细胞免疫应答,为布鲁氏菌表位疫苗的筛选与构建提供参考。

Study of the immune response characteristics of Brucella Omp2b dominant T-B combined antigen epitope

The dominant B cell epitopes and T cell epitopes of Omp2b protein were predicted and analyzed through bioinformatics technology, and T-B combined antigen epitopes were screened to explore their immunogenicity and immune response characteristics, thus laying a foundation for the development of an effective Brucella vaccine. The bioinformatics software programs ProtParam, SOMPA, Swiss-model, Rasmol, DNAStar, SYFPEITHI and IEDB were used to analyze the structure of the Omp2b protein and predict the dominant epitopes of T cells and B cells, as well as the T-B combined epitopes. The number of IFN-γ positive cells was detected with the ELISPOT method. ELISA was used to detect the specific IgG antibody level of Omp2b T-B combined peptide in the serum of patients with brucella infection. In vitro culture of immune cells was performed, and perforin and granulase B were detected in the supernatant. Bioinformatics software was used to comprehensively analyze Omp2b protein, and a potential segment at positions 196-216 of the T-B cell combined antigen epitope was screened out. The number of IFN-γ cells (SFU) secreted in the patient group was significantly higher than that in the healthy control group (F=25.413,P<0.01). Serum levels of IgG antibodies, perforin and granulase B in patients with brucellosis were also significantly elevated (F=13.653,P<0.01). The T-B combined epitope of Omp2b protein produces a specific humoral immune response and cellular immune response, thus providing a reference for the selection and construction of a Brucella epitope vaccine.