목차

1.Introduction 1
1.1 Vaccine adjuvants 1
1.2 Influenza virus 10
1.3 Influenza vaccine and adjuvants 16
2. Materials and methods 23
2.1 Ethic 23
2.2 Reagents 23
2.3 Influenza vaccine and adjuvants 24
2.4 Cell lines and culture conditions 24
2.5 Influenza virus strains and virus propagation 24
2.6 Determination of influenza virus titers 25
2.6.1 Determination of hemagglutination units of influenza virus preparations 25
2.6.2 Determination of 50% tissue culture infectious dose of influenza virus preparations 27
2.7 Immunization of mice 27
2.8 Measurement of serum IgG antibody titer 28
2.9 Measurement of serum hemagglutination-inhibition antibody titers 29
2.10 Virus neutralization assay 31
2.11 Cytokine ELISA 33
2.12 Mouse protection assay 33
2.13 Measurement of virus titers in the mouse lung tissues 34
2.14 Histology of mouse lungs 35
2.15 RT-PCR 35
2.16 Statistical analysis 38
3.Results 39
3.1 Preparation of influenza virus 39
3.2 CIA06 increases serum antibody titers to influenza vaccine Greenflu-S® 41
3.3 CIA06-adjuvanted Greenflu-S® induce antibodies with higher cross-reactivity against heterologous H1N1 virus strains 48
3.4 CIA06-adjuvanted Greenflu-S® induces Th1-type-prodominant immune responses 50
3.5 CIA06 promotes the protective efficacy of Greenflu-S® against lethal infection with influenza virus 54
3.6 Persistence of the protective immunity elicited by the CIA06-adjuvanted influenza vaccine 59
3.7 Safety of CIA06-adjuvanted Greenflu-S® against excessive inflammatory response upon viral infection 63
3.8 Application of CIA06 to trivalent influenza subunit HA vaccine 68
3.9 Application of CIA06 to avian influenza recombinant HA vaccine 74
4.Discussion 79
1.Introduction 86
1.1 Pseudomonas aeruginosa infection 86
1.2 P. aeruginosa vaccine and adjuvant 91
2.Materials and methods 95
2.1 Reagents 95
2.2 Cell lines and culture conditions 95
2.3 Characterization of P. aeruginosa strains 96
2.3.1 P. aeruginosa strains and culture conditions 96
2.3.2 LPS O-antigen serotyping of P. aeruginosa strains 96
2.3.3 Determination of in vitro cytotoxicity of P. aeruginosa strains 98
2.3.4 Determination of 50% lethal dose of P. aeruginosa strains 98
2.4 P. aeruginosa OMP antigens and adjuvants 99
2.5 Immunization of mice 99
2.6 Measurement of antigen-specific serum IgG antibody titers 101
2.7 Opsonophagocytosis assay 102
2.7.1 Preparation of fluorescence-labeled P. aeruginosa 102
2.7.2 Differentiation of HL-60 cells 102
2.7.3 Measurement of opsonophagocytic activity of mouse immune sera 103
2.8 Mouse protection assay 105
2.9 Statistical analysis 105
3.Results 106
3.1 Preparation of P. aeruginosa strains 106
3.2 CIA06 increase serum IgG antibody titers to P. aeruginosa OMP antigen 112
3.3 Antibodies induced by CIA06-adjuvanted vaccine exhibit increased cross-reactivity to heterologous serotype strains 117
3.4 Antibodies induced by CIA06-adjuvanted vaccine exhibit increased opsonophagocytic activity 120
3.5 CIA06 promote the protective efficacy of the vaccine against P. aeruginosa infection 125
3.6 CIA06 enhances the immunogenicity and protective efficacy of the FT1 vaccine 127
4.Discussion 131
5.References 134
6.국문초록 149