title: Yeast-surface expressed BVDV E2 protein induces a Th1/Th2 response in naïve T cells.
abstract: Yeast species such as Saccharomyces cerevisiae are known to be potent activators of the immune system. S. cerevisiae activates the innate immune system by engaging pattern recognition receptors such as toll like receptor 2 (TLR2) and dectin-1. In the current project, we express the immunogenic envelope protein E2 of bovine viral diarrhoea virus (BVDV) on the surface of S. cerevisiae. After successful expression, components of the innate and adaptive immune response induced by the recombinant S. cerevisiaein vitro were analysed to determine if expression in yeast enhances the immunogenicity of the viral protein. Recombinant S. cerevisiae stimulated production of the chemokine CXCL-8 in primary bovine macrophages, but did no stimulate production of reactive oxygen species (ROS) in the same cells. Additionally, bovine macrophages primed with S. cerevisiae expressing viral envelope proteins had a greater capacity for stimulating proliferation of CD4+ T-cells from BVDV-free animals compared to macrophages primed with envelope protein alone or S. cerevisiae without envelope protein expression. Heat inactivation of recombinant S. cerevisiae increased ROS production and capacity to stimulate CD4+ T-cells in macrophages but did not alter CXCL-8 release compared to the live counter-part. Additionally, heat-inactivation of recombinant S. cerevisiae induced less INFγ and IL-4 but equal amounts of IL-10 compared to live yeast T-cell cultures. Our studies demonstrate a use for S. cerevisiae as a vehicle for transporting BVDV vaccine antigen to antigen-presenting cell in order to elicit cell-mediated immunity even in naïve animals.
authors: Royal Veterinary College, Department of Pathology and Infectious Diseases, Hawkshead Lane, Hatfield AL9 7TA, UK.
authors: Robert
authors: R
authors: Patterson
authors: Jessica
authors: J
authors: Nerren
authors: Michael
authors: M
authors: Kogut
authors: Pinar
authors: P
authors: Court
authors: Bernardo
authors: B
authors: Villarreal-Ramos
authors: Hans-Martin
authors: HM
authors: Seyfert
authors: Paul
authors: P
authors: Dalby
authors: Dirk
authors: D
authors: Werling
mesh_terms: D000818:Animals; D002417:Cattle; D049109:Cell Proliferation; D002478:Cells, Cultured; D001908:Diarrhea Viruses, Bovine Viral / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D030781:Organisms, Genetically Modified; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D018417:Th1 Cells / Q000276:immunology / Q000382:microbiology / Q000502:physiology*; D018418:Th2 Cells / Q000276:immunology / Q000382:microbiology / Q000502:physiology*; D022223:Vaccines, Subunit; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014765:Viral Vaccines / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D016209:Interleukin-8; D008565:Membrane Proteins; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D022223:Vaccines, Subunit; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C076551:gp53, bovine viral diarrhea virus; D015847:Interleukin-4; D007371:Interferon-gamma