Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 308, Issue 1-2, Pages 216-230Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2005.11.005
Keywords
lymphocyte responses; highly active antiretroviral therapy (HAART); interferon gamma; perforin; interleukin-4; T-cell memory; T-cell anergy
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Funding
- MRC [G0501957] Funding Source: UKRI
- Medical Research Council [G0501957] Funding Source: Medline
- Wellcome Trust Funding Source: Medline
- Medical Research Council [G0501957] Funding Source: researchfish
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The enzyme-linked inummospot (ELIspot) assay is a highly sensitive and valuable tool for determining the frequency of cytokine-secreting T cells. It is essential to determine both frequencies and functional capabilities of antigen-specific T cells, including cytokine secretion, degranulation, and cytotoxicity in order to obtain a faller picture of the immune status of an individual. We describe here for the first time a perforin-release ELIspot assay which, when used in combination with IFN-gamma and IL-4 ELIspots, permits rapid assessment of these functional parameters for antigen-specific T cells. Whole antigen or peptides from HIV-1, recall and other viral antigens were used for in vitro stimulation. Anti-HIV-1 responses in treated chronically infected individuals were weak, both in terms of perform and IFN-gamma production. Tetanus toxoid stimulation was associated with moderate perforin release and a predominantly type-2 IL-4 producing response, whilst herpes simplex virus antigen stimulation resulted in perforin release but only a weak type-1 IFN-gamma response. Anti-cytomegalovirus responses generated high levels of perform in conjunction with IFN-gamma. Cytokines IL-2 and IL-12/IL-15 induced perform release coupled with an IFN-gamma type-1 response. Perform release strongly correlated with IFN-gamma production to individual influenza, Epstein-Barr virus or cytomegalovirus MHC class I restricted peptides, in an HIV-1 sero-negative cohort, indicating a cytolytic type-1 CD8(+) T-cell response. Evaluation of immunogenicity and putative efficacy of candidate vaccines using IFN-gamma will not be as informative alone as when combined with perforin and IL-4 evaluations, which allow assessment of specific cytotoxic potential without extensive cell culture. (c) 2005 Elsevier B.V. All rights reserved.
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