title: A prime/boost DNA/Modified vaccinia virus Ankara vaccine expressing recombinant Leishmania DNA encoding TRYP is safe and immunogenic in outbred dogs, the reservoir of zoonotic visceral leishmaniasis.
abstract: Previous studies demonstrated safety, immunogenicity and efficacy of DNA/modified vaccinia virus Ankara (MVA) prime/boost vaccines expressing tryparedoxin peroxidase (TRYP) and Leishmania homologue of the mammalian receptor for activated C kinase (LACK) against Leishmania major challenge in mice, which was consistent with results from TRYP protein/adjuvant combinations in non-human primates. This study aimed to conduct safety and immunogenicity trials of these DNA/MVA vaccines in dogs, the natural reservoir host of Leishmania infantum, followed-up for 4 months post-vaccination. In a cohort of 22 uninfected outbred dogs, blinded randomised administration of 1000 microg (high dose) or 100 microg (low dose) DNA prime (day 0) and 1x10(8)pfu MVA boost (day 28) was shown to be safe and showed no clinical side effects. High dose DNA/MVA vaccinated TRYP dogs produced statistically higher mean levels of the type-1 pro-inflammatory cytokine IFN-gamma than controls in whole blood assays (WBA) stimulated with the recombinant vaccine antigen TRYP, up to the final sampling at day 126, and in the absence of challenge with Leishmania. TRYP vaccinated dogs also demonstrated significantly higher TRYP-specific total IgG and IgG2 subtype titres than in controls, and positive in vivo intradermal reactions at day 156 in the absence of natural infection, observed in 6/8 TRYP vaccinated dogs. No significant increases in IFN-gamma in LACK-stimulated WBA, or in LACK-specific IgG levels, were detected in LACK vaccinated dogs compared to controls, and only 2/9 LACK vaccinated dogs demonstrated DTH responses at day 156. In all groups, IgG1 subclass responses and antigen-specific stimulation of IL-10 were similar to controls demonstrating an absence of Th2/T(reg) response, as expected in the absence of in vivo restimulation or natural/experimental challenge with Leishmania. These collective results indicate significant antigen-specific type-1 responses and in vivo memory phase cellular immune responses, consistent with superior potential for protective vaccine immunogenicity of DNA/MVA TRYP over LACK.
authors.affiliation: Populations and Disease Research Group, Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK. c.carson@warwick.ac.uk
authors.forename: Connor
authors.initials: C
authors.lastname: Carson
authors.forename: Maria
authors.initials: M
authors.lastname: Antoniou
authors.forename: Maria Begoña
authors.initials: MB
authors.lastname: Ruiz-Argüello
authors.forename: Antonio
authors.initials: A
authors.lastname: Alcami
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Christodoulou
authors.forename: Ippokratis
authors.initials: I
authors.lastname: Messaritakis
authors.forename: Jenefer M
authors.initials: JM
authors.lastname: Blackwell
authors.forename: Orin
authors.initials: O
authors.lastname: Courtenay
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology; D004285:Dogs; D007117:Immunization, Secondary / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000378:metabolism; D018314:Leishmania infantum / Q000276:immunology; D054332:Leishmaniasis Vaccines / Q000008:administration & dosage / Q000009:adverse effects* / Q000276:immunology*; D007898:Leishmaniasis, Visceral / Q000276:immunology / Q000517:prevention & control*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D010544:Peroxidases / Q000235:genetics / Q000276:immunology; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000009:adverse effects* / Q000276:immunology*; D014616:Vaccinia virus / Q000235:genetics
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D054332:Leishmaniasis Vaccines; D015800:Protozoan Proteins; D019444:Vaccines, DNA; C093139:LACK antigen, Leishmania; D007371:Interferon-gamma; D010544:Peroxidases; C110959:tryparedoxin peroxidase