猪带绦虫TSOL18重组耻垢分枝杆菌疫苗诱导小鼠免疫应答

Immune responses in mice induced with a TSOL18 recombinant Mycobacterium smegmatis vaccine of Taenia solium

目的观察猪带绦虫TSOL18重组耻垢分枝杆菌(Mycobacterium smegmatis,MS)疫苗免疫小鼠诱导产生的体液和细胞免疫应答。方法SPF级昆明小鼠60只(18~22 g,雌雄各半),采用随机数字表法分为3组,每组20只,分别为重组MS-TSOL18疫苗组、MS对照组、磷酸盐缓冲液(PBS)对照组。采用口服灌胃法免疫2次,间隔2周。于免疫后0、2、4、6、8周,每周4只小鼠眼眶静脉取血,分离血清,采用酶联免疫吸附试验(ELISA)法检测小鼠血清IgG与IgG2a水平;无菌取脾,制备脾淋巴细胞悬液,采用CCK-8法检测脾淋巴细胞经特异性抗原刺激后增殖水平,采用ELISA法检测小鼠脾淋巴细胞经特异性抗原刺激后产生白细胞介素(IL)-2和IL-4的水平。结果与MS对照组(IgG:0.160 ± 0.019、0.187 ± 0.038、0.193 ± 0.050、0.170 ± 0.005,IgG2a:0.213 ± 0.010、0.198 ± 0.012)和PBS对照组(IgG:0.159 ± 0.015、0.184 ± 0.029、0.191 ± 0.025、0.165 ± 0.018,IgG2a:0.198 ± 0.032、0.178 ± 0.025)比较,重组MS-TSOL18疫苗组小鼠血清特异性IgG水平(0.310 ± 0.034、0.391 ± 0.029、0.443 ± 0.030、0.373 ± 0.021)在免疫后2、4、6、8周升高(P均< 0.05),lgG2a水平(0.446 ± 0.056、0.339 ± 0.026)在免疫后6、8周升高(P均< 0.05),6周时均达峰值。抗原刺激后,与MS对照组和PBS对照组比较,重组MS-TSOL18疫苗组小鼠脾淋巴细胞增殖水平在免疫后2、4、6、8周升高(P均< 0.05),在6周时达峰值。抗原刺激后,与MS对照组和PBS对照组比较,重组MS-TSOL18疫苗组小鼠脾淋巴细胞培养上清液中IL-2和IL-4水平均在免疫后2、4、6、8周升高(P均< 0.05),分别在6周和4周时达峰值。结论猪带绦虫TSOL18重组MS疫苗免疫可诱导小鼠产生体液和细胞免疫应答。

Objective To observe humoral and cellular immune responses induced in mice by a TSOL18 recombinant Mycobacterium smegmatis (MS) vaccine of Taenia solium. Methods Sixty specific pathogen-free (SPF) Kunming mice (18-22 g, half male and half female) were divided into 3 groups by random number table method, 20 mice in each group. One group was administered with recombinant MS-TSOL18 vaccine, one group was administered with MS as control, and one group was administered phosphate buffer saline (PBS) as control. Kunming mice were vaccinated once every two weeks for 2 times through intragastric administration. At 0, 2, 4, 6, and 8 weeks after immunization, blood sample was collected and serum was separated. The levels of IgG and IgG2a in serum were detected using enzyme linked immunosorbent assay (ELISA). The spleen was separated to prepare spleen suspension. The proliferation of spleen lymphocyte with the stimulation of a specific antigen was detected by CCK-8. The levels of interleukin (IL)-2 and IL-4 in spleen cell culture supernatant with the stimulation of a specific antigen were detected by ELISA. Results Compared with MS control group (IgG: 0.160 ± 0.019, 0.187 ± 0.038, 0.193 ± 0.050, 0.170 ± 0.005;IgG2a: 0.213 ± 0.010, 0.198 ± 0.012) and PBS control group (IgG: 0.159 ± 0.015, 0.184 ± 0.029, 0.191 ± 0.025, 0.165 ± 0.018;IgG2a: 0.198 ± 0.032, 0.178 ± 0.025), the levels of specific IgG (0.310 ± 0.034, 0.391 ± 0.029, 0.443 ± 0.030, 0.373 ± 0.021) in recombinant MS-TSOL18 vaccine group increased at 2, 4, 6, and 8 weeks after immunization (P < 0.05), the levels of specific IgG2a (0.446 ± 0.056, 0.339 ± 0.026) in recombinant MS-TSOL18 vaccine group increased at 6 and 8 weeks after immunization (P < 0.05), and reached the highest level by the 6th week. After antigen stimulation, compared with MS and PBS control groups, the levels of spleen lymphocyte proliferation increased at 2, 4, 6, and 8 weeks after immunization (P < 0.05), and reached the highest level by the 6th week. After antigen stimulation, compared with MS and PBS control groups, the levels of IL-2 and IL-4 in spleen lymphocyte culture supernatant increased at 2, 4, 6, and 8 weeks after immunization (P < 0.05), and reached the highest level by the 6th and 4th weeks, respectively. Conclusion The recombinant MS-TSOL18 vaccine of Taenia solium might induce mice to produce humoral and cellular immune responses.