CpG-ODN对流感病毒裂解疫苗免疫应答的影响

发布时间：2018-03-27 09:07

本文选题：流感病毒　切入点：裂解疫苗　出处：《西北农林科技大学》2011年硕士论文

【摘要】：虽然目前广泛使用的流感疫苗能够较为有效地预防和控制流感流行,但仍存在流感大流行时疫苗产能不足和季节性流感病毒疫苗免疫效果有待改善等问题。疫苗佐剂的合理使用为有效地解决这些问题提供了思路。本论文研究了新型疫苗佐剂CpG-ODN对流感病毒裂解疫苗体液免疫和细胞免疫效果的影响,为今后研制新佐剂流感疫苗奠定了基础。在本研究中以2009 H1N1流感病毒裂解疫苗为抗原,分别以CpG-ODN、CpG-ODN和氢氧化铝复合佐剂以及CpG-ODN和ISA720复合佐剂为疫苗佐剂,通过肌肉注射和滴鼻免疫BALB/c小鼠,通过ELISA、血凝抑制试验和假病毒中和试验等方法评价体液免疫效果,通过ELISPOT、胞内细胞因子染色和体内CTL杀伤等方法评价细胞免疫效果。肌肉免疫结果表明,CpG-ODN单佐剂能够在一定程度上增强体液免疫,2针免疫后不同抗原剂量组中抗原特异性IgG抗体滴度、血凝抑制抗体滴度和中和抗体滴度分别提高3～6倍、2～4倍和4～8倍。CpG-ODN和氢氧化铝复合佐剂具有更强的佐剂效应,2针免疫后不同抗原剂量组中抗原特异性IgG抗体滴度、血凝抑制抗体滴度和中和抗体滴度分别提高23～57倍、9～20倍和16～64倍。根据体液免疫结果,CpG-ODN和氢氧化铝复合佐剂能够使流感病毒裂解疫苗的抗原用量降低至少16倍。CpG-ODN单独使用时并不能增强流感病毒裂解疫苗的细胞免疫应答,而CpG-ODN和氢氧化铝复合佐剂不但能够促进抗原特异性CD4~+ T细胞的IFN-γ分泌,而且能够促进抗原特异性CD8~+ T细胞的CTL杀伤活性。黏膜免疫结果表明,CpG-ODN单佐剂能够增强流感病毒裂解疫苗的体液免疫应答,2次免疫后血清中抗原特异性IgG抗体滴度与无佐剂组相比提高10倍,并且血清和黏膜局部均可检测到抗原特异性IgA抗体。同时,CpG-ODN还能够在一定程度上增强细胞免疫应答,不但能够促进抗原特异性CD4~+ T细胞的IFN-γ分泌,而且能够促进抗原特异性CD8~+ T细胞CTL杀伤活性。与CpG-ODN单佐剂相比,CpG-ODN和氢氧化铝复合佐剂以及CpG-ODN和ISA720复合佐剂并未显示出更强的佐剂效应。综上所述, CpG-ODN单独作为流感病毒裂解疫苗黏膜佐剂能够诱生黏膜局部免疫、系统性体液免疫和细胞免疫,而CpG-ODN和氢氧化铝复合佐剂作为流感病毒裂解疫苗肌肉免疫佐剂能够明显增强体液免疫和细胞免疫应答并显著降低抗原用量。
[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- 纬 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- 纬 secretion of antigen-specific CD4T cells, but also to promote the secretion of IFN- 纬 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.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

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