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.
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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)
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Copyright | Copyright © 2011 Elsevier B.V. All rights reserved. |
Chemical References |
- Antigens, Viral
- Immunodominant Epitopes
- Viral Proteins
- Brefeldin A
- Interferon-gamma
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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)
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