Title Page
ABSTRACT
Contents
Introduction 12
Materials and methods 18
Virus and cell 18
Expression and purification of ENC-VP8* and P2-VP8* 20
Characterization of ENC-VP8*and P2-VP8* 25
Binding of ENC-VP8* and P2-VP8* to synthesized oligosaccharides 25
Vaccine formulation and mice immunization 26
Determination of antigen-specific antibody responses 28
Antibody response inhibiting the binding with synthesized oligosaccharides 28
Plaque reduced neutralization antibody response 29
Virus-specific T-cell response 30
Results 31
Characterization of ENC-VP8* and P2-VP8* 31
VP8* specific antibodies response of the monovalent vaccine 36
VP8* specific antibodies response of the trivalent vaccine 48
VP8*-Led H type 1 binding inhibitory antibody response of the monovalent vaccine 55
VP8*-Led H type 1 binding inhibitory antibody response of the trivalent vaccine 63
Rotavirus neutralization antibody response of the monovalent vaccine 67
Rotavirus neutralization antibody response of the trivalent vaccine 75
Virus-specific T-cell response of the monovalent vaccine 80
Virus-specific T-cell response of the trivalent vaccine 84
Discussion 87
논문개요 94
Reference 96
Table 1. Three serotypes of rotavirus A used in this study 19
Table 2. Amino acid sequence of rotavirus VP8* 23
Table 3. Amino acid sequence of encapsulin 24
Table 4. G1P[8]VP8* specific IgG response of G1P[8]VP8* monovalent vaccine immunized sera 39
Table 5. G2P[4]VP8* specific IgG response of G2P[4]VP8* monovalent vaccine immunized sera 42
Table 6. G2P[6]VP8* specific IgG response of G2P[6]VP8* monovalent vaccine immunized sera 45
Table 7. VP8* specific IgG response of the trivalent vaccine immunized sera 51
Table 8. BD50 titer of G1P[8]VP8* monovalent vaccine immunized sera[이미지참조] 58
Table 9. BD50 titer of G2P[4]VP8* monovalent vaccine immunized sera[이미지참조] 60
Table 10. BD50 titer of G2P[6]VP8* monovalent vaccine immunized sera[이미지참조] 62
Table 11. BD50 titer of the trivalent vaccine immunized sera[이미지참조] 66
Table 12. PRNT50 titer of G1P[8]VP8* monovalent vaccine immunized sera[이미지참조] 70
Table 13. PRNT50 titer of G2P[4]VP8* monovalent vaccine immunized sera[이미지참조] 72
Table 14. PRNT50 titer of G2P[6]VP8* monovalent vaccine immunized sera[이미지참조] 74
Table 15. PRNT50 titer of the trivalent vaccine immunized sera[이미지참조] 79
Fig. 1. Structure of human rotavirus A 17
Fig. 2. Vector construct for expression of encapsulin conjugated VP8* (ENC-VP8*) 22
Fig. 3. Schedule of mice experiment. 27
Fig. 4. SDS PAGE analysis of ENC-VP8* 32
Fig. 5. SDS PAGE analysis of P2-VP8* 33
Fig. 6. Morphological analysis of ENC-VP8* under transmission electron microscopy. 34
Fig. 7. HBGA binding analysis of ENC-VP8* and P2-VP8* 35
Fig. 8. G1P[8]VP8* specific IgG response of G1P[8]VP8* monovalent vaccine immunized individual mice sera 40
Fig. 9. G1P[8]VP8* specific IgA response of G1P[8]VP8* monovalent vaccine 41
Fig. 10. G2P[4]VP8* specific IgG response of G2P[4]VP8* monovalent vaccine immunized individual mice sera 43
Fig. 11. G2P[4]VP8* specific IgA response of G2P[4]VP8* monovalent vaccine 44
Fig. 12. G2P[6]VP8* specific IgG response of G2P[6]VP8* monovalent vaccine immunized individual mice sera 46
Fig. 13. G2P[6]VP8* specific IgA response of G2P[6]VP8* monovalent vaccine 47
Fig. 14. VP8* specific IgG response of the trivalent vaccine immunized individual mice sera 52
Fig. 15. VP8* specific IgA response of the trivalent vaccine 54
Fig. 16. G1P[8]VP8*-Led H type 1 binding inhibitory response of G1P[8]VP8* monovalent vaccine 57
Fig. 17. G2P[4]VP8*-Led H type 1 binding inhibitory response of G2P[4]VP8* monovalent vaccine 59
Fig. 18. G2P[6]VP8*-Led H type 1 binding inhibitory response of G2P[6]VP8* monovalent vaccine 61
Fig. 19. VP8*-Led H type 1 binding inhibitory response of the trivalent vaccine 65
Fig. 20. G1P[8] virus neutralizing antibody response of G1P[8]VP8* monovalent vaccine 69
Fig. 21. G2P[4] virus neutralizing antibody response of G2P[4]VP8* monovalent vaccine 71
Fig. 22. G2P[6] virus neutralizing antibody response of G2P[6]VP8* monovalent vaccine 73
Fig. 23. Three serotypes of rotavirus neutralizing antibody response of the trivalent vaccine 77
Fig. 24. T-cell response of G1P[8]VP8* monovalent vaccine 81
Fig. 25. T-cell response of G2P[4]VP8* monovalent vaccine 82
Fig. 26. T-cell response of G2P[6]VP8* monovalent vaccine 83
Fig. 27. T-cell response of the trivalent vaccine 85