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梅毒是一种由梅毒螺旋体(Treponema pallidum,Tp)引起的临床表现复杂的性传播疾病,近年来世界各国包括中国的梅毒发病率呈逐年上升趋势,研制有效疫苗预防该种疾病已成为各国关注的公共卫生问题.DNA疫苗具有免疫效果好且持久、无毒力回复等特点成为近年来研究热点.对于Tp核酸疫苗研究,国内外除了本课题组构建TpDNA疫苗免疫新西兰兔进行免疫活性的初步研究外,尚未见Tp核酸疫苗的相关研究.本研究旨在探讨评价白细胞介素-2(interleukin-2,IL-2)与Tp膜蛋白GpdDNA疫苗用壳聚糖(chitosan,CS)纳米颗粒包裹联合免疫新西兰兔后对Tp皮肤感染后的保护效应.将真核表达重组体pcDNA3.1(+)/IL-2和pcDNA3.1(+)/TpGpd采用壳聚糖纳米颗粒包裹或未包裹方式处理后分别或联合免疫新西兰兔,每隔2周加强1次共免疫3次,并于初次免疫后第10周各实验组兔皮下接种Tp标准株进行背部皮肤感染实验,观察记录感染早期各实验组皮损的变化情况,在免疫及感染期间不同时间点ELISA检测免疫兔特异性抗体产生水平和脾细胞IL-2及IFN-诱导水平,MTT法检测兔脾细胞增殖水平.初步结果证实,pcD/IL-2基因佐剂可以显著加强pTpGpd疫苗在免疫期(0~8周)兔体内的特异性抗体及脾细胞增殖分化水平以及维持Tp感染后168天时间内长期稳定的高抗体水平及高脾细胞增殖分化水平.pcD/IL-2基因佐剂和CS纳米颗粒的联合使用对pcD/TpGpd疫苗的抗Tp皮肤感染的保护性效应达到最佳.
Syphilis is a complicated sexually transmitted disease caused by Treponema pallidum (Tp). In recent years, the incidence of syphilis in all countries in the world, including China, has been on the rise. The development of effective vaccines to prevent this disease has become a national concern Public health issues.DNA vaccine has a good immune effect and lasting, non-toxic response and other characteristics have become the research hotspot in recent years.For Tp nucleic acid vaccine research, in addition to our group to build TpDNA vaccine immunized New Zealand rabbit for preliminary study of immune activity , No Tp DNA vaccines have been reported.The aim of this study is to evaluate the efficacy and safety of the combination of chitosan (CS) nanoparticles and interleukin-2 (IL-2) and Tp membrane protein GpdDNA vaccine Protective effect of New Zealand rabbits on Tp skin infection.Eukaryotic recombinant pcDNA3.1 (+) / IL-2 and pcDNA3.1 (+) / TpGpd were treated with chitosan nanoparticles or uncoated New Zealand rabbits were immunized separately or co-immunized once every 2 weeks for 3 times. Tp standard strains were subcutaneously inoculated subcutaneously in the rabbits at the 10th week after the initial immunization to test the skin infection on the back. The changes of skin lesions in the early experimental groups were recorded. The levels of IL-2 and IFN-induced IL-2 and IFN-induced immune rabbit antibodies were detected by ELISA at different time points during immunization and infection. The proliferation of rabbit splenocytes was detected by MTT assay. The preliminary results confirmed that adjuvant pcD / IL-2 could significantly enhance the proliferation and differentiation of pTpGpd vaccines during the immunization period (0-8 weeks) in rabbits and the long-term stability within 168 days after Tp infection High antibody level and the proliferation and differentiation of high spleen cells.Combined use of pcDNA3.1 / IL-2 gene adjuvant and CS nanoparticle achieved the best protective effect on pcD / TpGpd vaccine against Tp skin infection.