Epitopes described in "Development of an interferon-gamma ELISPOT assay to detect human T cell responses to HSV-2."

Article Authors:Christine M Posavad; Amalia S Magaret; Lin Zhao; Dawn E Mueller; Anna Wald; Lawrence Corey
Article Title:Development of an interferon-gamma ELISPOT assay to detect human T cell responses to HSV-2.
Reference Detail
Reference ID:1023027
Abstract:BACKGROUND: The need for an HSV-2 vaccine is great considering the increasing prevalence of HSV-2 despite the widespread use of antiviral drugs. Human clinical trials of HSV-2 vaccines that elicit neutralizing antibodies have proven to be only partially effective suggesting that induction of effective T cell responses to HSV-2 is also a critical component to an efficacious vaccine. A sensitive and specific assay to measure HSV-specific T cell responses is a necessary part of vaccine development and thus we undertook the development of an interferon- (IFN-) ELISPOT assay to measure T cell responses to HSV-2. METHODS: PBMC from HSV-seronegative (HSVneg) (n=35), HSV-1-seropositive (HSV-1+/2-) (n=20) and HSV-2-seropositive (HSV-2+) subjects (n=26) were screened by IFN- ELISPOT for T cell responses using 34 peptide pools representing 16 HSV-2 proteins including mostly virion and immediate-early (IE) proteins. RESULTS: Overall, 85% of HSV-2+ subjects had a positive response to the HSV-2 peptide pools and on average, HSV-2+ subjects responded to 3 peptide pools (range 1-10). The most frequent responses were to gD-2, UL39, UL46, ICP0, UL49, gB-2, and ICP4. In contrast, only 2 of 35 (6%) HSVneg subjects had detectable T cell responses and in both cases, responses were of low magnitude relative to responses in HSV-2+ subjects and were directed at a single peptide pool. The response rate to the HSV-2 peptide pools in HSV-1+/2- subjects was 40% suggesting that the HSV-2 peptide pools contain a significant number of type-common T cell epitopes. The IFN- ELISPOT assay detected CD4 and CD8 T cells directed at HSV-2 peptides as confirmed by intracellular cytokine staining and flow cytometry. CONCLUSION: We have developed a quantitative IFN- ELISPOT assay that detects both CD4 and CD8 T cells to HSV-2 peptides. This assay does not require large quantities of PBMC to generate dendritic cells for T cell stimulation, making it an ideal assay for monitoring the immunogenicity of candidate HSV-2 vaccines designed to elicit T cell responses to HSV-2 specific epitopes.
Affiliations:Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA. posavad@uw.edu.
Reference Type:Literature
PubMed ID:21801778
Journal Volume:29
Article Pages:7058-66
Journal ISSN:0264-410X
Article Chemical List:gov.nih.nlm.ncbi.www.jaxb.impl.NameOfSubstanceImpl@71720b7;gov.nih.nlm.ncbi.www.jaxb.impl.NameOfSubstanceImpl@7f64d203;gov.nih.nlm.ncbi.www.jaxb.impl.NameOfSubstanceImpl@3418044c;gov.nih.nlm.ncbi.www.jaxb.impl.NameOfSubstanceImpl@14c2344c;gov.nih.nlm.ncbi.www.jaxb.impl.NameOfSubstanceImpl@59f77439
Article MeSH List:Adult; CD4-Positive T-Lymphocytes(immunology); CD8-Positive T-Lymphocytes(immunology); Enzyme-Linked Immunospot Assay(methods); Epitopes, T-Lymphocyte(immunology); Female; Herpes Simplex Virus Vaccines(immunology); Herpesvirus 1, Human(immunology); Herpesvirus 2, Human(immunology); Humans; Immediate-Early Proteins(immunology); Interferon-gamma(immunology); Male; Middle Aged; Peptides(immunology); Phenotype
Curation Last Updated:2015-01-18 20:14:28