分子佐剂Ov-ASP-1对弓形虫SAG1核酸疫苗的影响
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摘要
为了研究分子佐剂Ov-ASP-1对弓形虫表面抗原糖蛋白1(SAG1)核酸疫苗的免疫效力的影响,用含有分子佐剂Ov-ASP-1的pVAX1-Ov-ASP-1-SAG1质粒、对照质粒pVAX1-SAG1、空载体pVAX1免疫鸡,并设置PBS对照组,通过抗体水平与细胞因子水平的检测结果判断免疫效果和免疫应答情况;然后用弓形虫RH株人工感染试验鸡,通过实时荧光定量PCR检测鸡的肺脏、脾脏、肝脏中的弓形虫数量,判断分子佐剂Ov-ASP-1是否增强鸡对弓形虫的抵抗力。结果表明:与其他组相比,试验各阶段pVAX1-Ov-ASP-1-SAG1组免疫球蛋白G(IgG)、白细胞介素-10(IL-10)、白细胞介素-12(IL-12)和干扰素γ(IFN-γ)水平都较高,差异达到了显著或极显著水平(P<0.05或P<0.01);攻虫后pVAX1-Ov-ASP-1-SAG1组感染弓形虫的鸡荷虫量比其他组极显著降低(P<0.01)。说明分子佐剂Ov-ASP-1可以显著增强弓形虫表面膜蛋白SAG1的免疫原性和免疫保护性。
In order to study the effects of molecular adjuvant Ov-ASP-1 on the immune efficacy of SAG1 protein of Toxoplasma gondii(T.gondii), chikens were divided into four groups(pVAX1-Ov-ASP-1-SAG1 group, pVAX1-SAG1 group, pVAX1 group and PBS control group), then the antibody and cytokines levels were determined to indicate the immune effects and response. Then the chickens were artificially infected with T. gondii RH strain.Real-time fluorescence quantitative PCR was used to determine the number of toxoplasma in lung, spleen, and liver of chickens, to determine whether the molecular adjuvant-Ov-ASP-1 chould enhance the resistance of chickens to T. gondii. Results showed that antibody IgG level and cytokines IL-10, IL-12, and IFN-γ were higher in the pVAX1-Ov-ASP1-SAG1 group than that in other groups(P<0.05 or P<0.01). The amount of parasites in chickens infected with T. gondii in pVAX1-Ov-ASP1-SAG1 group was extremly lower than that in other groups(P<0.01). It indicates that the molecular adjuvant Ov-ASP-1 can significantly enhance the immunogenicity and immunoprotection of SAG1 protein of T. gondii.
In order to study the effects of molecular adjuvant Ov-ASP-1 on the immune efficacy of SAG1 protein of Toxoplasma gondii(T.gondii), chikens were divided into four groups(pVAX1-Ov-ASP-1-SAG1 group, pVAX1-SAG1 group, pVAX1 group and PBS control group), then the antibody and cytokines levels were determined to indicate the immune effects and response. Then the chickens were artificially infected with T. gondii RH strain.Real-time fluorescence quantitative PCR was used to determine the number of toxoplasma in lung, spleen, and liver of chickens, to determine whether the molecular adjuvant-Ov-ASP-1 chould enhance the resistance of chickens to T. gondii. Results showed that antibody IgG level and cytokines IL-10, IL-12, and IFN-γ were higher in the pVAX1-Ov-ASP1-SAG1 group than that in other groups(P<0.05 or P<0.01). The amount of parasites in chickens infected with T. gondii in pVAX1-Ov-ASP1-SAG1 group was extremly lower than that in other groups(P<0.01). It indicates that the molecular adjuvant Ov-ASP-1 can significantly enhance the immunogenicity and immunoprotection of SAG1 protein of T. gondii.
引文
[1] CHING X T, FONG M Y, LAU Y L. Evaluation of the protective effect of deoxyribonucleic acid vaccines encoding granule antigen 2 and 5 against acute toxoplasmosis in Balb/c mice[J]. Am J Trop Med Hyg, 2017, 96(6):1441-1447.
[2] ZHU W N, WANG J L, KAI C, et al. Evaluation of protective immunity induced by DNA vaccination with genes encoding Toxoplasma gondii, GRA17 and GRA23 against acute toxoplasmosis in mice[J]. Exp Parasitol, 2017, 179:20-27.
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[4] 刘海阔, 彭高辉, 袁子国, 等.抗弓形虫病核酸疫苗的研究进展[J]. 中国人兽共患病学报,2009,25(10):1004-1007.
[5] SHANG L M, LIU Q, LIU W S, et al. Protection in mice immunized with a Heterologous prime-boost regime using DNA and recombinant pseudorabies expressing Tg SAG1 against Toxoplasma gondii. challenge [J]. Vaccine,2010,27:2741-2745.
[6] BATISTA-DUHARTE A, TELLEZ-MARTINEZ D, FUENTES DLP, et al. Molecular adjuvants that modulate regulatory T cell function in vaccination: a critical appraisal[J]. Pharmacol Res,2018, 129:237-250.
[7] SUSCHAK J J, WILLIAMS J A, SCHMALJOHN C S. Advancements in DNA vaccine vectors, non-mechanical delivery methods, and molecular adjuvants to increase immunogenicity[J]. Hum Vacc Immunother, 2017, 13(12):2837-2848.
[8] MACDONALD A J, TAWE W, LEON O, et al. Ov-ASP-1,the Onchocerca volvulus homologue of the activation associated secreted protein family is immunostimulatory and can induce protective anti-larval immunity[J]. Parasite Immunol,2004,26(l):53-62.
[9] MACDONALD A J, CAO L, HE Y X, et al. rOv-ASP-1, a recombinant secreted protein of the helminth Onchocerca volvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens[J]. Vaccine,2005,23(26):3446-3352.
[10] 何帅, 周春雪, 朱兴全,等. 弓形虫不同发育时期棒状体颈部蛋白5基因的表达研究[J]. 中国畜牧兽医, 2017, 44(2):350-356.
[11] 侯照峰, 王尚尚, 贾红,等. 弓形虫荧光定量PCR检测方法的建立与初步应用[J]. 中国兽医杂志, 2015, 51(1):3-6.
[12] 杜海娟. 弓形虫pVAX1-SAG1和pVAX1-SAG4基因疫苗的构建及动物免疫保护性的研究[D]. 南宁:广西医科大学, 2014.
[2] ZHU W N, WANG J L, KAI C, et al. Evaluation of protective immunity induced by DNA vaccination with genes encoding Toxoplasma gondii, GRA17 and GRA23 against acute toxoplasmosis in mice[J]. Exp Parasitol, 2017, 179:20-27.
[3] HAN Y, ZHOU A, GANG L, et al. Protection via a ROM4 DNA vaccine and peptide against Toxoplasma gondii, in Balb/c mice[J]. Bmc Infect Dis, 2017, 17(1):59.
[4] 刘海阔, 彭高辉, 袁子国, 等.抗弓形虫病核酸疫苗的研究进展[J]. 中国人兽共患病学报,2009,25(10):1004-1007.
[5] SHANG L M, LIU Q, LIU W S, et al. Protection in mice immunized with a Heterologous prime-boost regime using DNA and recombinant pseudorabies expressing Tg SAG1 against Toxoplasma gondii. challenge [J]. Vaccine,2010,27:2741-2745.
[6] BATISTA-DUHARTE A, TELLEZ-MARTINEZ D, FUENTES DLP, et al. Molecular adjuvants that modulate regulatory T cell function in vaccination: a critical appraisal[J]. Pharmacol Res,2018, 129:237-250.
[7] SUSCHAK J J, WILLIAMS J A, SCHMALJOHN C S. Advancements in DNA vaccine vectors, non-mechanical delivery methods, and molecular adjuvants to increase immunogenicity[J]. Hum Vacc Immunother, 2017, 13(12):2837-2848.
[8] MACDONALD A J, TAWE W, LEON O, et al. Ov-ASP-1,the Onchocerca volvulus homologue of the activation associated secreted protein family is immunostimulatory and can induce protective anti-larval immunity[J]. Parasite Immunol,2004,26(l):53-62.
[9] MACDONALD A J, CAO L, HE Y X, et al. rOv-ASP-1, a recombinant secreted protein of the helminth Onchocerca volvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens[J]. Vaccine,2005,23(26):3446-3352.
[10] 何帅, 周春雪, 朱兴全,等. 弓形虫不同发育时期棒状体颈部蛋白5基因的表达研究[J]. 中国畜牧兽医, 2017, 44(2):350-356.
[11] 侯照峰, 王尚尚, 贾红,等. 弓形虫荧光定量PCR检测方法的建立与初步应用[J]. 中国兽医杂志, 2015, 51(1):3-6.
[12] 杜海娟. 弓形虫pVAX1-SAG1和pVAX1-SAG4基因疫苗的构建及动物免疫保护性的研究[D]. 南宁:广西医科大学, 2014.