阳离子脂质体DOTAP佐剂对H5N1流感病毒裂解疫苗免疫效果的影响

发布时间：2018-08-09 18:06

【摘要】：目的研究阳离子脂质体DOTAP佐剂对H5N1型流感病毒裂解疫苗免疫效果的影响。方法制备DOTAP阳离子脂质体流感病毒裂解疫苗(简称DOTAP流感裂解疫苗),检测其包封率。将BALB/c小鼠分为13组,分别用含0.1、1.0、10.0μg HA/只剂量以DOTAP、Al(OH)3、CPG-ODN为佐剂以及不含佐剂的流感裂解疫苗于0、21天皮下免疫,PBS作为对照组,用血凝抑制试验检测小鼠初次免疫后21、42天血清HI抗体滴度;用ELISA检测初次免疫后21、42天血清特异性IgG抗体、IgG1、IgG2a亚类抗体滴度,以及初次免疫后42天小鼠脾脏单个核细胞体外经抗原刺激后细胞因子IL-2、IL-4、IFN-γ的分泌水平。将BALB/c小鼠分为3组,分别用含不同DOTAP剂量(100、300、600μg/只)的DOTAP流感裂解疫苗于0、21天皮下免疫,检测初次免疫后21、42天小鼠血清HI抗体滴度和IgG抗体滴度。结果 DOTAP流感裂解疫苗粒径在300~400 nm,带正电荷,包封率在50%以上;DOTAP流感裂解疫苗诱导的HI抗体水平和特异性IgG抗体水平均高于流感裂解疫苗,而与铝佐剂和Cp G-ODN佐剂间差异无统计学意义;DOTAP流感裂解疫苗产生的抗体仍以IgG1亚类抗体为主,免疫后42天诱导的IgG2a亚类抗体水平高于流感裂解疫苗和铝佐剂,低于Cp G-ODN佐剂;DOTAP流感裂解疫苗免疫后既分泌高水平Th1型细胞因子IFN-γ,同时也分泌高水平Th2型细胞因子IL-4;不同DOTAP剂量的DOTAP流感裂解疫苗免疫后,其HI抗体滴度和IgG抗体滴度在低、中、高剂量组之间存在明显的量效关系。结论 DOTAP作为H5N1型流感病毒裂解疫苗的佐剂可显著提高流感裂解疫苗的免疫原性,其对体液免疫应答的增强作用不低于铝佐剂和Cp G-ODN佐剂,并具有诱导细胞免疫应答的能力。
[Abstract]:Objective to study the effect of cationic liposome DOTAP adjuvant on immune response of H5N1 influenza virus lytic vaccine. Methods DOTAP cationic liposome influenza virus lytic vaccine (DOTAP vaccine) was prepared and its entrapment efficiency was determined. BALB/c mice were divided into 13 groups. BALB/c mice were subcutaneously immunized with dotapride Al (OH) 3G ODN as adjuvant and influenza cleavage vaccine without adjuvant for 21 days. Hemagglutination inhibition test was used to detect HI antibody titer of serum 2142 days after primary immunization. ELISA was used to detect the titer of serum specific IgG antibody IgG2a on day 21 and 42 after primary immunization, as well as the level of cytokine IL-2mIL-4 and IFN- 纬 in splenic mononuclear cells (SMC) stimulated by antigen on day 42 after primary immunization. BALB/c mice were subcutaneously immunized with DOTAP vaccine containing different doses of DOTAP (100300600 渭 g / mouse) for 21 days. The serum HI antibody titer and IgG antibody titer were detected 21 ~ 42 days after the first immunization. Results the HI antibody level and specific IgG antibody level induced by DOTAP influenza lytic vaccine were higher than those of influenza lytic vaccine with positive charge and encapsulation efficiency of more than 50%. However, there was no significant difference between the vaccine and aluminum adjuvant and CP G-ODN adjuvant. The antibody produced by DOTAP influenza cleavage vaccine was still dominated by IgG1 subclass antibody. The level of IgG2a subclass antibody induced 42 days after immunization was higher than that of influenza cleavage vaccine and aluminum adjuvant. Lower than CP G-ODN adjuvant dotap influenza lytic vaccine not only secreted high level of Th1 cytokine IFN- 纬, but also secreted high level of Th2 cytokine IL-4.The HI antibody titer and IgG antibody titer of DOTAP influenza lytic vaccine with different DOTAP dosages were low. There was a significant dose-effect relationship between medium and high dose groups. Conclusion DOTAP, as an adjuvant of H5N1 influenza virus lytic vaccine, can significantly improve the immunogenicity of influenza lytic vaccine, and its effect on humoral immune response is no less than that of aluminum adjuvant and CP G-ODN adjuvant, and it has the ability to induce cellular immune response.
【作者单位】：武汉生物制品研究所有限责任公司病毒性疫苗研究二室;
【基金】：国家重大新药创制项目(2014ZX09102045-011)
【分类号】：R392

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