Identification of an immunodominant CD4+ T cell epitope in the VP6 protein of rotavirus following intranasal immunization of BALB/c mice.
The only lymphocytes required for protection against fecal rotavirus shedding after intranasal immunization of BALB/c (H-2(d)) mice with a chimeric rotavirus VP6 protein (MBPColon, two colonsVP6) and the mucosal adjuvant LT(R192G) are CD4(+) T cells. The purpose of this study was to identify CD4(+) T cell epitopes within VP6 that might be responsible for this protection. To make this determination, spleen cells obtained from BALB/c mice following intranasal immunization with MBPColon, two colonsVP6/LT(R192G) were stimulated in vitro with either MBPColon, two colonsVP6 or overlapping VP6 peptides containing <or=30 amino acids (AA). The numbers of memory (CD44(high)) CD4(+) T cells stimulated to produce T(H)1 and T(H)17 cytokines (IFNgamma and IL-17), as well as the quantities of these cytokines released into the cell supernatants, were then measured relative to those produced in mock-stimulated cells from the same animals. One epitope expected to be found was the VP6 14-mer AA(289-302), previously identified as a CD4(+) T cell epitope in H-2(d) mice. This was not observed but instead the only VP6 epitope identified was AA(242-259), the dominant CD4(+) T cell epitope previously reported after oral, live rotavirus immunization.
McNeal
Monica M
MM
Division of Infectious Diseases, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Basu
Mitali
M
Bean
Judy A
JA
Clements
John D
JD
Choi
Anthony H-C
AH
Ward
Richard L
RL
D000281:Administration, Intranasal; D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D036022:Capsid Proteins / Q000276:immunology*; D002478:Cells, Cultured; D016207:Cytokines / Q000032:analysis / Q000096:biosynthesis; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005260:Female; D007111:Immunity, Cellular; D007115:Immunization Schedule; D016056:Immunodominant Epitopes / Q000276:immunology; D018655:Lymphocyte Count; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000008:administration & dosage / Q000138:chemical synthesis / Q000235:genetics; D012401:Rotavirus / Q000276:immunology*; D012400:Rotavirus Infections / Q000517:prevention & control*; D022243:Rotavirus Vaccines / Q000008:administration & dosage*; D013154:Spleen / Q000276:immunology; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000008:administration & dosage
D000956:Antigens, Viral; D036022:Capsid Proteins; D016207:Cytokines; D018984:Epitopes, T-Lymphocyte; D016056:Immunodominant Epitopes; D010455:Peptides; D022243:Rotavirus Vaccines; C061332:VP6 protein, Rotavirus; D014614:Vaccines, Synthetic