【作者】 王亚军；

【导师】 盛军；

【作者基本信息】 吉林大学 ， 微生物与生化药学， 2006， 硕士

【摘要】 为了获得更加经济、高效、安全的流感疫苗,本实验室以脂质体为佐剂包封流感疫苗,制得流感脂质体疫苗。以氢化大豆卵磷脂和胆固醇为膜材制得流感脂质体疫苗并对其包封率、粒径、及其形态进行了全面的检测。用脂质体流感疫苗对Balb/c小鼠进行单针腹腔免疫,分别用血凝抑制试验(HI)和酶联免疫吸附试验(ELISA)检测免后几周的小鼠血清的抗体水平;MTT法检测脾淋巴细胞增殖比例;流式细胞术检测小鼠T淋巴细胞表面标记;ELISA试剂盒检测小鼠肺内容物中细胞因子IL-4、INFγ水平;在免后12周用活毒A3(H3N2EID50=10-8.0)对小鼠进行攻击一周后检测小鼠肺部病变,测得疫苗对小鼠机体的保护效率。实验结果表明脂质体流感疫苗可激发更强的体液免疫,在脾淋巴细胞增殖比例,CD4水平和CD4/CD8比例,以及细胞因子(IL-4、IFNγ)水平均上均有显著的增强作用,能够对机体产生时间更长,更高效的保护作用。本研究表明脂质体流感疫苗与普通无佐剂流感疫苗相比,具有明显的优势,这种具有双重免疫效果的疫苗必将在对高危人群的保护上具有更大的潜力。更多还原

【Abstract】 Objective: to improve the potency of the currently used influenzasplit vaccine, which are of relatively low efficiency in high-riskgroups. To produce liposome-encapsulated influenza splitvaccine(H3N2) and analyzing its humoral and cellular immuneresponse elicited in mice.Method: 1、produce liposome-encapsulated influenza split vaccineand its characterization was studied;2、 then a single dose wasadministered i.p. to BALB/c mice (5-6 per group) with nonliposomalor liposomal vaccines. Serum antibodies were assayed on weeks 2-13by the haemagglutination-inhibition(HI) test and ELISA. 3、UsingMTT to measure the spleen cell proliferation in vitro;to determinethe levels of CD4 and CD8 T-cell and the CD4/CD8 using flowcytometry;to quantify the levels of IL-4 and IFNγ in lunghomogenates by ELISA kit;4、the measurement of protectiveimmunity was assessed by the prevention of lung injury from livevirus 12weeks post vaccination.Results: the following results were obtained :(1) The efficiencyof encapsulation in liposomes was 91.1%.(2)Following immunizationwith 2μg and 4μg liposome-encapsulated influenza split vaccine,there was a higher, up to 4fold stronger HI titer and ELISA titer thanthat obtained with non-liposomal HN.(3)The combined liposomalvaccines consisting of encapsulated antigen, but not the nonliposomalvaccine, elicited a higher titer of he spleen cell proliferation and cellsubsets like CD4+ and CD4+/CD8+ and the liposome-encapsulatedinfluenza split vaccine triggered cytokine (IL-4 and IFNγ)production.(4)The level of protection following vaccination with thecombined liposomal vaccines was 4/6 versus 2/6 in mice immunizedwith non-liposomal vaccine.Conclusion: our animal experiments show that the liposomalvaccines are superior to the currently used influenza vaccines,increasing the response by 2-4folds in mice, stimulating both TH1and TH2 responses. Such vaccines may be more potent in high-riskgroup than the currently used split vaccines.更多还原