Title Page
Contents
Overall Abstract 18
Ⅰ. Overall Introduction 21
1. Vaccine 21
2. Immune systems in the elderly 30
3. Adjuvant 36
4. Single-stranded RNA adjuvant 44
5. Purpose of the present study 47
Ⅱ. CHAPTER 1. Effective inactivated influenza vaccine for the elderly using a viral IRES based single-stranded RNA adjuvant 48
ⅰ. Abstract 49
ⅱ. Introduction 50
ⅲ. Materials and Methods 54
ⅳ. Results 69
ⅴ. Discussion 94
ⅵ. References 99
Ⅱ. CHAPTER 2. Nanostructures of single-stranded RNA adjuvant enhance the efficacy of conventional split inactivated influenza vaccine 104
ⅰ. Abstract 105
ⅱ. Introduction 107
ⅲ. Materials and Methods 111
ⅳ. Results 124
ⅴ. Discussion 155
ⅵ. References 161
Ⅲ. Overall Discussion 166
Ⅳ. References of Overall Introduction and Discussion 174
국문 초록 189
Table 1. Licensed adjuvants for human use and type of immune response. 43
Table 2. List of viral IRES. Two viral IRESes were used for ssRNA adjuvant platform. 126
Figure 1. The primary and secondary immune responses. 22
Figure 2. Various vaccine platforms. 25
Figure 3. Immunological changes associated with aging. 32
Figure 4. Ideal condition of an adjuvant. 37
Figure 5. Inactivated influenza vaccine (IIV) formulated with ssRNA augments the IIV-induced humoral responses. BALB/c mice were... 73
Figure 6. Inactivated influenza vaccine (IIV) formulated with ssRNA reduces viral replication and pathogenesis and induces protection against... 78
Figure 7. Inactivated influenza vaccine (IIV) formulated with ssRNA increases the generation of influenza HA-specific T cells and humoral... 83
Figure 8. Inactivated influenza vaccine (IIV) formulated with ssRNA increases antigen-presenting cells, activation markers, and cytokine... 88
Figure 9. Immune-related gene expression pattern in splenocytes of immunized young and aged mice. Differentially expressed genes (DEGs)... 92
Figure 10. Circular Dichroism (CD) spectroscopy analysis of G4 motif. (A) A schematic representation of the ssRNA adjuvant platform, CrPV... 127
Figure 11. Comparison of the dendritic cell activation efficacy between ssRNA adjuvant candidates with or without G4 structure in human... 130
Figure 12. Comparison of immunostimulatory responses of ssRNA adjuvant candidates with or without G4 structure in vivo. BALB/c mice... 133
Figure 13. Comparison of SINV-1G and AddaVax-induced humoral immune responses. BALB/c mice were intramuscularly immunized with... 137
Figure 14. Comparison of SINV-1G and AddaVax-induced T cell responses. BALB/c mice were intramuscularly immunized with 1 μg of... 141
Figure 15. Gene expression profiles of IIV-formulated with or without SINV-1G and AddaVax in lymph node and spleen. BALB/c mice were... 149
Figure 16. The immune cell subset alterations in the spleen. BALB/c mice were intramuscularly immunized with 1 μg of IIV either alone or with... 153
Figure 17. Schematic of T cells and B cells in immunity induced by SINV-1G and AddaVax. 154