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: Nelson B Moseley, Oskar Laur, Chris C Ibegbu, Gilbert D Loria, Gini Ikwuenzunma, Himangi R Jayakar, Michael A Whitt, John D Altman
• Journal: Journal of immunological methods (J Immunol Methods) Vol. 375 Issue 1-2 Pg. 118-28 (Jan 31 2012) ISSN: 1872-7905 [Electronic] Netherlands
• PMID: 22004852 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2011 Elsevier B.V. All rights reserved.

Chemical References

• Antigens, Viral
• Immunodominant Epitopes
• Viral Proteins
• Brefeldin A
• Interferon-gamma

Topics

• Animals
• Antigens, Viral (immunology)
• Brefeldin A (immunology)
• Cells, Cultured
• Cricetinae
• Cytomegalovirus (immunology)
• DNA Replication (immunology)
• Dendritic Cells (immunology)
• Genetic Vectors (genetics, immunology)
• Humans
• Immunodominant Epitopes (immunology)
• Interferon-gamma (immunology)
• Lymphocyte Activation (immunology)
• Monocytes (immunology)
• Orthomyxoviridae (immunology)
• T-Lymphocytes, Cytotoxic (immunology)
• T-Lymphocytes, Helper-Inducer (immunology)
• Vesicular stomatitis Indiana virus (genetics, immunology, physiology)
• Viral Proteins (genetics, immunology)
• Virus Replication
• Yellow fever virus (immunology)