不同佐剂与恶性疟疾疫苗M.RCAg-1相配伍后的免疫原性研究

发布时间：2018-01-07 20:08

本文关键词：不同佐剂与恶性疟疾疫苗M.RCAg-1相配伍后的免疫原性研究　出处：《北京协和医学院》2016年硕士论文　论文类型：学位论文

【摘要】：疟疾作为世界三大传染病之一仍然严重威胁着人类的健康。据世界卫生组织统计,2015年在全球大约有32亿人(约占世界总人口的一半)面临着感染疟疾的风险,其中大约有2.14亿人患病,43.8万人死亡。近几年,由于疟原虫耐药性虫株的不断出现,以及介导疟原虫传播的蚊媒对杀虫剂逐渐产生抗性,使得一些传统的针对恶性疟疾的防控措施和手段都面临着严峻的挑战。对于控制和消除疟疾最有希望和最有效的工具就是引进一种高度有效的疟疾疫苗。根据相关文献报道,恶性疟原虫红内期疫苗多种候选抗原单独或简单的联合使用时,其免疫效果并不理想,因此构建多抗原多表位的重组蛋白疫苗成为红内期疟疾疫苗研究的方向之一。本课题组前期通过“表位重组”技术构建了疟原虫随机重组抗原-1(malaria random constructed antigen-1, M.RCAg-1),将其作为恶性疟疾红内期多表位人工随机重组蛋白抗原M.RCAg-1在BALB/c小鼠、新西兰兔、非人灵长类动物模型中证实其具有良好的免疫原性和体内外免疫保护性,显示其具有研发前景。通过与中国科学院过程工程研究所的合作,已研发出稳定的纯化工艺,建立了稳定简便可行的中试规模制备工艺。本研究在前期工作基础上选用了在临床上经常用于治疗骨质疏松的药物-XX'性膦酸盐-ALD、ZLD和PMD与M.RCAg-1相配伍,并在BALB/c小鼠模型和新西兰兔体内评价其免疫效果,进而筛选出最佳佐剂和免疫剂量,为今后进行的M.RCAg-1的临床前研究奠定基础。目前主要取得以下进展：1.从与M.RCAg-1以不同剂量和不同免疫方式相配伍的双性膦酸盐免疫BALB/c小鼠,确定佐剂ZLD (0.05mg s.c.)组为最佳配伍剂量组。并通过免疫新西兰兔,获得疟原虫体外生长抑制率最高达76.7%。2.从与M.RCAg-1以不同剂量相配伍的口服佐剂ALD免疫BALB/c小鼠,确定ALD(0.1mg)组为最佳配伍剂量组。并通过免疫新西兰兔,获得疟原虫体外生长抑制率最高达48.6%。3.从与M.RCAg-1以不同剂量相配伍的ZLD和MF59联合免疫BALB/c小鼠,确定ZLD (0.05mg)+MF59组为最佳配伍剂量组,同时证明其两者联合不仅能够增强其Th1型免疫应答,同时能够增强其Th2型免疫应答。并通过免疫新西兰兔,获得疟原虫体外生长抑制率最高达72.5%。
[Abstract]:Malaria as one of the world's three major infectious diseases are still a serious threat to human health. According to the WHO statistics, in 2015 about 3 billion 200 million people (about half of the world's total population) facing the risk of infection, there are about 214 million people in which 438 thousand people died. In recent years, because of drug resistance strains of Plasmodium the emergence of malaria transmission and mediated mosquito resistance to insecticides gradually, making some traditional for malaria prevention and control measures and means are facing severe challenges. For malaria control and elimination the hope and the most effective tool is the introduction of a highly effective malaria vaccine. According to the relevant literature Plasmodium falciparum, a variety of vaccine candidate antigens alone or in combination with the use of simple, the immune effect is not ideal, so the construction of multi heavy group protein multi epitope antigen The vaccine has become one of the direction of blood stage malaria vaccine research. Ourprevious by epitope recombinant technology constructs the Plasmodium random recombinant antigen -1 (malaria random constructed antigen-1, M.RCAg-1), as the malaria blood stage polyepitope artificial random group protein antigen M.RCAg-1 in BALB/c mice and New Zealand rabbits. Confirmed the good immunogenicity and protective immunity in vitro and in vivo animal models of nonhuman primates, shows that it has good prospects for development. Through cooperation with China Academy of Sciences Institute of process engineering, has developed a process of purification and stability, to establish a stable and feasible pilot scale preparation process. This study in the early stage on the basis of the work used in clinical drug frequently used to -XX'phosphonate -ALD in the treatment of osteoporosis, ZLD and PMD and M.RCAg-1 compatible, and in a mouse model of BALB/c and New Zealand rabbits In vivo evaluation of the immune effect, and then selected the best adjuvant and immunization dose, lay the foundation for future clinical research of M.RCAg-1. The following is the main achievement: 1. from the double phosphonate immune BALB/ and M.RCAg-1 with different dose and different immunization methods compatible c mice, ZLD (0.05mg s.c.) to determine the adjuvant group as the optimal dosage group. And through the immune rabbit to obtain the growth inhibition rate of up to 76.7%.2. from M.RCAg-1 with different dose combined oral adjuvant ALD BALB/c mice were immunized with Plasmodium in vitro, determination of ALD (0.1mg) group for the optimal dosage group. By immunizing the rabbit to obtain the growth inhibition rate of up to 48.6%.3. from the M.RCAg-1 with different doses of combined ZLD and MF59 combined with BALB/c mice were immunized with Plasmodium falciparum in vitro, determination of ZLD (0.05mg) group +MF59 was the best dosage group, and prove the combination It can not only enhance its Th1 type immune response, but also enhance its Th2 immune response. By immunizing New Zealand rabbits, the growth rate of malaria parasites is up to 72.5%..

【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R392-33

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