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用不同的佐剂与高致病性人禽流感H5N1全病毒疫苗混合,通过透皮途径免疫BALB/c小鼠并评价其免疫应答效果,从而初步筛选出较好的透皮免疫佐剂。实验选用CT、CpG ODN1826、CpG ODN2006、MF59四种不同的佐剂按适当的比例与高致病性人禽流感H5N1灭活全病毒抗原混合制成透皮疫苗,透皮免疫BALB/c小鼠,检测血清IgG抗体效价、血清中和抗体效价,以及肺、鼻灌洗液中特异性IgG和IgA抗体效价,并对脾淋巴细胞分泌的IFN-γ和IL-4细胞因子水平进行评估。结果显示,在CpG ODN1826单独使用或与CT合用组血清IgG抗体效价较无佐剂组均有显著升高(P<0.05),其中尤以CpG1826+CT+HA组血清IgG抗体效价最高,同时血清中和抗体效价也验证了这一点,肺、鼻灌洗液中,均可检测到特异性IgA、IgG。佐剂组与无佐剂组、PBS组、空白对照组相比较,脾淋巴细胞分泌IFN-γ和IL-4的水平均见提高(P<0.05),并且CpG1826+CT+HA组与滴鼻组相比无统计学意义(P=0.2267)。结果表明,在高致病性人禽流感H5N1病毒透皮疫苗的小鼠模型中,CT与CpG ODN1826是较好的的透皮免疫佐剂,能够诱导机体Th2型和Th1型免疫应答,同时诱导了黏膜免疫应答的产生,这为进一步研究提供了基础。
BALB / c mice were immunized with different adjuvants and the HPAI H5N1 virus. The transdermal immunization of BALB / c mice was evaluated and the immune responses were evaluated. The better transdermal immune adjuvants were screened out. The experiment selected CT, CpG ODN1826, CpG ODN2006, MF59 four kinds of different adjuvants in the appropriate ratio and highly pathogenic human H5N1 inactivated whole virus virus mixed antigens made transdermal vaccine, transdermal immunization BALB / c mice The serum IgG antibody titers, serum neutralizing antibody titers, and specific IgG and IgA antibody titers in lung and nasal lavage fluid were measured. The levels of IFN-γ and IL-4 cytokines secreted by splenic lymphocytes Evaluation. The results showed that the serum IgG antibody titers of CpG ODN1826 alone or in combination with CT were significantly higher than those in the non-adjuvant group (P <0.05), and the serum IgG antibody titer of CpG1826 + CT + HA group was the highest, Serum neutralizing antibody titer also verified this point, lung, nasal lavage, can detect specific IgA, IgG. The levels of IFN-γ and IL-4 secreted by splenic lymphocytes in both adjuvant group and non-adjuvant group were significantly increased (P <0.05), compared with those in control group, PBS group and blank control group Group compared with no significant (P = 0.2267). The results showed that CT and CpG ODN1826 were better transdermal immune adjuvants in the mouse model of HPAI H5N1 transmissible vaccine and could induce the induction of Th2 and Th1 immune responses The mucosal immune response, which provides the basis for further research.