title: Use of replication restricted recombinant vesicular stomatitis virus vectors for detection of antigen-specific T cells.
abstract: Detection of antigen-specific T cells at the single-cell level by ELISpot or flow cytometry techniques employing intracellular cytokine staining (ICS) is now an indispensable tool in many areas of immunology. When precisely mapped, optimal MHC-binding peptide epitopes are unknown, these assays use antigen in a variety of forms, including recombinant proteins, overlapping peptide sets representing one or more target protein sequences, microbial lysates, lysates of microbially-infected cells, or gene delivery vectors such as DNA expression plasmids or recombinant vaccinia or adenoviruses expressing a target protein of interest. Here we introduce replication-restricted, recombinant vesicular stomatitis virus (VSV) vectors as a safe, easy to produce, simple to use, and highly effective vector for genetic antigen delivery for the detection of human antigen-specific helper and cytotoxic T cells. To demonstrate the broad applicability of this approach, we have used these vectors to detect human T cell responses to the immunodominant pp65 antigen of human cytomegalovirus, individual segments of the yellow fever virus polyprotein, and to various influenza proteins.
authors: Emory Vaccine Center at the Yerkes National Primate Center at Emory University, United States.
authors: Nelson B
authors: NB
authors: Moseley
authors: Oskar
authors: O
authors: Laur
authors: Chris C
authors: CC
authors: Ibegbu
authors: Gilbert D
authors: GD
authors: Loria
authors: Gini
authors: G
authors: Ikwuenzunma
authors: Himangi R
authors: HR
authors: Jayakar
authors: Michael A
authors: MA
authors: Whitt
authors: John D
authors: JD
authors: Altman
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D020126:Brefeldin A / Q000276:immunology; D002478:Cells, Cultured; D006224:Cricetinae; D003587:Cytomegalovirus / Q000276:immunology; D004261:DNA Replication / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology*; D007371:Interferon-gamma / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D009000:Monocytes / Q000276:immunology; D009975:Orthomyxoviridae / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology; D014779:Virus Replication; D015005:Yellow fever virus / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D016056:Immunodominant Epitopes; D014764:Viral Proteins; D020126:Brefeldin A; D007371:Interferon-gamma