title: A modified cholera holotoxin CT-E29H enhances systemic and mucosal immune responses to recombinant Norwalk virus-virus like particle vaccine.
abstract: In this study, we evaluated the potential of a genetically modified cholera toxin, CT-E29H as an adjuvant for recombinant Norwalk virus like particle (NV-VLP) vaccine. This detoxified mutant, containing E to H substitution at amino acid 29 of the CT-A1 subunit, was administered with a recombinant Norwalk virus like particle vaccine to Balb/c mice by mucosal routes to monitor the induction of mucosal, humoral and cellular responses. We observed that a low dose of NV-VLP (5 microg) with the adjuvant delivered by the intranasal route (IN) was more effective than the highest dose (200 microg) delivered by oral route at inducing both cellular and NV-VLP specific IgG and IgA responses. Higher counts of antigen specific IgA secreting cells were observed in the Peyer's Patches (PP) following delivery of the vaccine with CT-E29H as compared to delivery of vaccine by mucosal routes without CT-E29H. Furthermore, there was an increase in antigen specific cells producing IL-4 from animals that received the vaccine with the adjuvant. Delivery of the vaccine by the oral route results in antigen specific CD4(+) and CD8(+) T cells in PP and spleen. Addition of CT-E29H results in an increase of antigen specific CD4(+) cell population in PP and both CD4(+) and CD8(+) populations in the spleen. These cellular and cytokine responses suggest that combining the vaccine with CT-E29H results in a stronger Th2 type response. Collectively, these results indicate that immune responses to NV-VLP vaccine are qualitatively and quantitatively improved when the vaccine is delivered along with CT-E29H, and thus merits its further consideration as a mucosal adjuvant.
authors.affiliation: Department of Viral Vaccine Immunology, Wyeth-Ayerst Research, Pearl River, NY 10965, USA.
authors.forename: Sangeeta B
authors.initials: SB
authors.lastname: Periwal
authors.forename: Kristin R
authors.initials: KR
authors.lastname: Kourie
authors.forename: Nandini
authors.initials: N
authors.lastname: Ramachandaran
authors.forename: Susan J
authors.initials: SJ
authors.lastname: Blakeney
authors.forename: Sylvia
authors.initials: S
authors.lastname: DeBruin
authors.forename: Duzhang
authors.initials: D
authors.lastname: Zhu
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Zamb
authors.forename: Larry
authors.initials: L
authors.lastname: Smith
authors.forename: Steve
authors.initials: S
authors.lastname: Udem
authors.forename: John H
authors.initials: JH
authors.lastname: Eldridge
authors.forename: Khushroo E
authors.initials: KE
authors.lastname: Shroff
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Reilly
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology*; D000281:Administration, Intranasal; D000818:Animals; D000917:Antibody Formation / Q000276:immunology; D002455:Cell Division / Q000187:drug effects; D002772:Cholera Toxin / Q000276:immunology* / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D018928:Immunity, Mucosal / Q000276:immunology*; D007070:Immunoglobulin A / Q000276:immunology; D007371:Interferon-gamma / Q000032:analysis; D015847:Interleukin-4 / Q000032:analysis / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects; D008221:Lymphoid Tissue / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D029322:Norovirus / Q000276:immunology*; D009663:Norwalk virus / Q000276:immunology*; D009924:Organ Culture Techniques; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D007070:Immunoglobulin A; D014614:Vaccines, Synthetic; D014765:Viral Vaccines; D015847:Interleukin-4; D007371:Interferon-gamma; D002772:Cholera Toxin; C476776:cholera holotoxin, His(29)-