CAJ | 학술논문

目的研究CpG-ODN和氢氧化铝复合佐剂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和解决流感疫苗产能不足的问题提供依据.方法以不同剂量的2009 H1N1流感病毒裂解疫苗为抗原,分别以CpG-ODN、氢氧化铝以及CpG-ODN和氢氧化铝复合佐剂为疫苗佐剂免疫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.结果与无佐剂对照组相比,CpG-ODN或氢氧化铝单独使用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,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.此外,复合佐剂能够显著增强流感病毒裂解疫苗的细胞免疫应答,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细胞的CTL杀伤活性.结论 CpG-ODN和氢氧化铝复合佐剂能够增强流感病毒裂解疫苗的体液免疫和细胞免疫应答并显著降低抗原用量.
목적 연구CpG-ODN화경양화려복합좌제대류감병독렬해역묘체액면역화세포면역효과적영향,위금후연제신좌제류감역묘화해결류감역묘산능불족적문제제공의거.방법 이불동제량적2009 H1N1류감병독렬해역묘위항원,분별이CpG-ODN、경양화려이급CpG-ODN화경양화려복합좌제위역묘좌제면역BALB/c소서,통과ELISA、혈응억제시험화가병독중화시험등방법평개체액면역효과,통과ELISPOT、포내세포인자염색화체내CTL살상등방법평개세포면역효과.결과 여무좌제대조조상비,CpG-ODN혹경양화려단독사용능구재일정정도상증강체액면역,2침면역후불동항원제량조중항원특이성IgG항체적도、혈응억제항체적도화중화항체적도분별제고3～6배、2～4배화4～8배.CpG-ODN화경양화려복합좌제구유경강적좌제효응,2침면역후불동항원제량조중항원특이성IgG항체적도、혈응억제항체적도화중화항체적도분별제고23～ 57배、9～20배화16～64배.근거체액면역결과,복합좌제능구사류감병독렬해역묘적항원용량강저지소16배.차외,복합좌제능구현저증강류감병독렬해역묘적세포면역응답,불단능구촉진항원특이성CD4+T세포적IFN-γ분비,이차능구촉진항원특이성CD8+T세포적CTL살상활성.결론 CpG-ODN화경양화려복합좌제능구증강류감병독렬해역묘적체액면역화세포면역응답병현저강저항원용량.
Objective To evaluate the immuno-potentiating effects of CpG-ODN plus alum as a composite adjuvant on influenza split virion vaccine.Methods BALB/c mice were immunized with various amounts of 2009 H1N1 influenza split virion vaccine,alone or in combination with CpG-ODN,alum,or both (composite adjuvant).Antigen-specific humoral immune responses were evaluated by ELISA,hemagglutination inhibiting (HI) assay and neutralizing assay.Antigen-specific cellular immune responses were evaluated by ELISPOT assay,intracellular cytokine staining assay and in vivo CTL assay.Results Compared with the control group immunized with antigen alone,a single use of either adjuvant weakly enhanced the humoral immune responses,as indicated by the increase of antigen-specific IgG titers,HI titers and neutralizing titers by 3-6 folds,2-4 folds and 4-8 folds,respectively,after two immunizations.In contrast,the composite adjuvant induced more potent humoral immune responses; the antigen-specific IgG titers,HI titers and neutralizing titers were increased by 23-57 folds,9-20 folds and 16-64 folds,respectively.Consequently,the composite adjuvant achieved antigen-sparing by at least 16 folds.In addition,the composite adjuvant significantly enhanced the antigen-specific cellular immune responses,as revealed by the increase of IFN-γ-secreting CD4+ T cells and the enhancement of CTL activity in immunized mice.Conclusion CpG-ODN plus alum as a composite adjuvant can enhance the immunogenicity of influenza split virion vaccine and achieve the antigen-sparing effect.