壳聚糖衍生物纳米颗粒在乙肝疫苗中的应用研究
发布时间：2018-08-30 19:44
【摘要】：我们新合成了多种壳聚糖的阳离子衍生物,并用其作为载体包裹重组非颗粒形式的乙肝HBcAg蛋白作为候选疫苗。为了评价其作为疫苗的可行性,以及诱导通过“内吞体至胞浆”途径形成交叉递呈的潜力,我们利用一系列试验对候选疫苗进行筛选,这些方法包括透射电镜、体外包裹及释放试验以及两个可以反映对内吞体的破坏能力的试验:体外的溶血试验和与RAW264.7共孵育后进行共聚焦显微镜观察。我们利用选取的壳聚糖衍生物(它们包括Ntrimethylaminoethylmethacrylate chloride—Methylmethacrylate(TM),Chitosan—Methylmethacrylate (CM),Chitosan—Ntrimethylaminoethylmethacrylate chloride—Methylmethacrylate(CTM) and Chitosan—N-dimethylaminoethylmethacrylate hydrochloride—Methylmethacrylate(CDM))包裹重组的乙肝HBcAg蛋白对小鼠进行免疫并且评价其免疫效果。与被接种纯蛋白溶液的组相比,这些接受候选疫苗免疫的小鼠被诱导出很强的特异性体液以及细胞免疫。在这些候选疫苗之中,CTM包裹的蛋白疫苗的免疫原性是最强的。从体内的免疫接种效果以及体外的一系列试验来看,通过纳米粒的包裹,候选疫苗可以通过“内吞体到胞浆”的途径来引起交叉递呈,同时也说明了此疫苗投放系统是大有潜力的。 诱导针对乙肝病毒抗原有效的细胞以及体液免疫的疫苗策略,是治疗慢性乙肝的关键。传统的核酸疫苗具有能有诱导体液免疫和细胞免疫的能力,然而它的效率不是很理想。为了提高相应核酸疫苗的效果,我们也研究了几种新合成的壳聚糖的阳离子衍生物作为纳米颗粒疫苗佐剂的应用情况。这些壳聚糖衍生物纳米粒在生理条件下显示更好的可溶性,增加了吸附亲水性分子的能力。这些纳米颗粒在体外可以保护DNA分子,延缓脱氧核糖核酸酶Ⅰ的分解作用。溶血试验和激光共聚焦显示这些纳米具有膜破坏活性因此具有潜在的加强基因转染的能力。我们用这些纳米粒去包裹pVAX—HBc的DNA疫苗,然后免疫C57BL/6小鼠,隔一周注射,共接
[Abstract]:We have synthesized several cationic derivatives of chitosan and used them as carriers to encapsulate non-granular hepatitis B HBcAg protein as candidate vaccine. To evaluate its feasibility as a vaccine and the potential for inducing cross-presentation through the endocytosomal to cytoplasmic pathway, we used a series of tests to screen candidate vaccines, including transmission electron microscopy. In vitro encapsulation and release tests and two tests reflecting the damage to endocytosis: hemolysis test in vitro and confocal microscopy after co-incubation with RAW264.7. The selected chitosan derivatives (including Ntrimethylaminoethylmethacrylate chloride-Methylmethacrylate (TM) Chitosan-Methylmethacrylate (CM) Chitosan-Ntrimethylaminoethylmethacrylate chloride-Methylmethacrylate (CTM) and Chitosan-N-dimethylaminoethylmethacrylate hydrochloride-Methylmethacrylate (CDM) were used to immunize mice and evaluate their immunological effects. Compared with those inoculated with pure protein solution, the mice immunized with the candidate vaccine were able to induce highly specific humoral and cellular immunity. Of these candidate vaccines, the CTM encased protein vaccine is the strongest immunogenicity. From the results of immunization in vivo and a series of experiments in vitro, through the encapsulation of nanoparticles, the candidate vaccine can cause cross-presentation through the "endocytosomal to cytoplasmic" pathway. It also shows that the vaccine delivery system has great potential. The key to the treatment of chronic hepatitis B is to induce effective cellular and humoral immunity against hepatitis B antigen. The traditional nucleic acid vaccine has the ability to induce humoral and cellular immunity, but its efficiency is not ideal. In order to improve the effect of nucleic acid vaccine, we also studied the application of several newly synthesized cationic derivatives of chitosan as adjuvant of nano-particle vaccine. These chitosan nanoparticles exhibited better solubility under physiological conditions and increased their ability to adsorb hydrophilic molecules. These nanoparticles can protect DNA molecules and delay the decomposition of deoxyribonuclease 鈪，
本文编号：2214100
[Abstract]:We have synthesized several cationic derivatives of chitosan and used them as carriers to encapsulate non-granular hepatitis B HBcAg protein as candidate vaccine. To evaluate its feasibility as a vaccine and the potential for inducing cross-presentation through the endocytosomal to cytoplasmic pathway, we used a series of tests to screen candidate vaccines, including transmission electron microscopy. In vitro encapsulation and release tests and two tests reflecting the damage to endocytosis: hemolysis test in vitro and confocal microscopy after co-incubation with RAW264.7. The selected chitosan derivatives (including Ntrimethylaminoethylmethacrylate chloride-Methylmethacrylate (TM) Chitosan-Methylmethacrylate (CM) Chitosan-Ntrimethylaminoethylmethacrylate chloride-Methylmethacrylate (CTM) and Chitosan-N-dimethylaminoethylmethacrylate hydrochloride-Methylmethacrylate (CDM) were used to immunize mice and evaluate their immunological effects. Compared with those inoculated with pure protein solution, the mice immunized with the candidate vaccine were able to induce highly specific humoral and cellular immunity. Of these candidate vaccines, the CTM encased protein vaccine is the strongest immunogenicity. From the results of immunization in vivo and a series of experiments in vitro, through the encapsulation of nanoparticles, the candidate vaccine can cause cross-presentation through the "endocytosomal to cytoplasmic" pathway. It also shows that the vaccine delivery system has great potential. The key to the treatment of chronic hepatitis B is to induce effective cellular and humoral immunity against hepatitis B antigen. The traditional nucleic acid vaccine has the ability to induce humoral and cellular immunity, but its efficiency is not ideal. In order to improve the effect of nucleic acid vaccine, we also studied the application of several newly synthesized cationic derivatives of chitosan as adjuvant of nano-particle vaccine. These chitosan nanoparticles exhibited better solubility under physiological conditions and increased their ability to adsorb hydrophilic molecules. These nanoparticles can protect DNA molecules and delay the decomposition of deoxyribonuclease 鈪，
本文编号：2214100
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