本文選題：流感病毒　切入點：裂解疫苗　出處：《西北農(nóng)林科技大學》2011年碩士論文

【摘要】：雖然目前廣泛使用的流感疫苗能夠較為有效地預防和控制流感流行,但仍存在流感大流行時疫苗產(chǎn)能不足和季節(jié)性流感病毒疫苗免疫效果有待改善等問題。疫苗佐劑的合理使用為有效地解決這些問題提供了思路。本論文研究了新型疫苗佐劑CpG-ODN對流感病毒裂解疫苗體液免疫和細胞免疫效果的影響,為今后研制新佐劑流感疫苗奠定了基礎(chǔ)。 在本研究中以2009 H1N1流感病毒裂解疫苗為抗原,分別以CpG-ODN、CpG-ODN和氫氧化鋁復合佐劑以及CpG-ODN和ISA720復合佐劑為疫苗佐劑,通過肌肉注射和滴鼻免疫BALB/c小鼠,通過ELISA、血凝抑制試驗和假病毒中和試驗等方法評價體液免疫效果,通過ELISPOT、胞內(nèi)細胞因子染色和體內(nèi)CTL殺傷等方法評價細胞免疫效果。 肌肉免疫結(jié)果表明,CpG-ODN單佐劑能夠在一定程度上增強體液免疫,2針免疫后不同抗原劑量組中抗原特異性IgG抗體滴度、血凝抑制抗體滴度和中和抗體滴度分別提高3～6倍、2～4倍和4～8倍。CpG-ODN和氫氧化鋁復合佐劑具有更強的佐劑效應,2針免疫后不同抗原劑量組中抗原特異性IgG抗體滴度、血凝抑制抗體滴度和中和抗體滴度分別提高23～57倍、9～20倍和16～64倍。根據(jù)體液免疫結(jié)果,CpG-ODN和氫氧化鋁復合佐劑能夠使流感病毒裂解疫苗的抗原用量降低至少16倍。CpG-ODN單獨使用時并不能增強流感病毒裂解疫苗的細胞免疫應答,而CpG-ODN和氫氧化鋁復合佐劑不但能夠促進抗原特異性CD4~+ T細胞的IFN-γ分泌,而且能夠促進抗原特異性CD8~+ T細胞的CTL殺傷活性。 黏膜免疫結(jié)果表明,CpG-ODN單佐劑能夠增強流感病毒裂解疫苗的體液免疫應答,2次免疫后血清中抗原特異性IgG抗體滴度與無佐劑組相比提高10倍,并且血清和黏膜局部均可檢測到抗原特異性IgA抗體。同時,CpG-ODN還能夠在一定程度上增強細胞免疫應答,不但能夠促進抗原特異性CD4~+ T細胞的IFN-γ分泌,而且能夠促進抗原特異性CD8~+ T細胞CTL殺傷活性。與CpG-ODN單佐劑相比,CpG-ODN和氫氧化鋁復合佐劑以及CpG-ODN和ISA720復合佐劑并未顯示出更強的佐劑效應。 綜上所述, CpG-ODN單獨作為流感病毒裂解疫苗黏膜佐劑能夠誘生黏膜局部免疫、系統(tǒng)性體液免疫和細胞免疫,而CpG-ODN和氫氧化鋁復合佐劑作為流感病毒裂解疫苗肌肉免疫佐劑能夠明顯增強體液免疫和細胞免疫應答并顯著降低抗原用量。
[Abstract]:Although currently widely used influenza vaccines are more effective in preventing and controlling influenza epidemics, However, there are still some problems such as insufficient vaccine capacity during influenza pandemic and the need to improve the immune effect of seasonal influenza virus vaccine. The rational use of vaccine adjuvant provides an effective way to solve these problems. The effect of CpG-ODN on humoral immunity and cellular immunity of influenza virus lytic vaccine. It lays a foundation for developing new adjuvant influenza vaccine in the future. In this study, 2009 H1N1 influenza virus lytic vaccine was used as antigen, CpG-ODN and aluminum hydroxide complex adjuvant and CpG-ODN and ISA720 complex adjuvant were used as vaccine adjuvant respectively. BALB/c mice were immunized by intramuscular injection and nasal drip. The humoral immune effect was evaluated by Elisa, hemagglutination inhibition test and pseudovirus neutralization test, and cellular immunity was evaluated by Elispot, intracellular cytokine staining and CTL killing in vivo. The results of muscle immunization showed that CpG-ODN single adjuvant could enhance the titer of antigen-specific IgG antibody in different dose groups after humoral immunization. The titer of hemagglutination inhibitory antibody and neutralizing antibody were increased by 3 ~ 6 times and 4 ~ 8 times, respectively. CpG-ODN and aluminum hydroxide complex adjuvant had stronger adjuvant effect on antigen-specific IgG antibody titers in different antigen groups after immunization. The titer of hemagglutination inhibitory antibody and neutralizing antibody were increased by 2357-fold, 920-fold and 1664-fold, respectively. According to the results of humoral immunity, CpG-ODN and aluminum hydroxide complex adjuvant could reduce the antigenic dosage of influenza virus lytic vaccine by at least 16 times. CpG-ODN alone. It does not enhance the cellular immune response of the influenza virus lytic vaccine. The compound adjuvant of CpG-ODN and aluminum hydroxide can not only promote IFN- 緯 secretion of antigen-specific CD4T cells, but also promote the CTL killing activity of antigen-specific CD8T cells. The results of mucosal immunity showed that CpG-ODN single adjuvant could enhance the humoral immune response of influenza virus lytic vaccine and the titer of antigen-specific IgG antibody in serum was increased by 10 times compared with that of no adjuvant group. Moreover, antigen-specific IgA antibodies can be detected in both serum and mucous membrane, and CpG-ODN can enhance cellular immune response to some extent, not only to promote IFN- 緯 secretion of antigen-specific CD4T cells, but also to promote the secretion of IFN- 緯 in antigen-specific CD4T cells. Compared with CpG-ODN single adjuvant, CpG-ODN, aluminum hydroxide complex adjuvant and CpG-ODN and ISA720 complex adjuvant did not show stronger adjuvant effect. To sum up, CpG-ODN alone as a mucosal adjuvant of influenza virus lysis vaccine can induce mucosal local immunity, systemic humoral immunity and cellular immunity. CpG-ODN and aluminum hydroxide complex adjuvant as muscle immune adjuvant of influenza virus lytic vaccine could significantly enhance humoral and cellular immune responses and significantly reduce the dosage of antigens.
【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R392

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