The ELISPOT assay: an easily transferable method for measuring cellular responses and identifying T cell epitopes.
Characterization of human leukocyte antigen (HLA) class I restricted epitopes derived from viral pathogens is imperative for formulating therapeutic interventions, as well as for vaccine design and monitoring. Sensitive, easy and cost-effective assays that measure the frequency of antigen-specific T lymphocytes are crucial for evaluating and improving vaccines and therapies. This paper reviews the ELISPOT technique that allows for quantifying HIV-specific T lymphocytes at the single cell level from peripheral blood by detection of antigen-induced cytokine secretion. The assay can be used successfully to quantify T cell immune responses in humans infected with different pathogens and to assess T cell immunogenicity of vaccines in phase I/II and III clinical trials. This review focuses on the ELISPOT methodology and discusses how it can be standardized and potentially used by multiple international laboratories attached to clinical trial sites.
Mashishi
Tumelo
T
AIDS Research Unit, National Institute for Communicable Diseases, Johannesburg, South Africa.
Gray
Clive M
CM
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D015497:HIV-1 / Q000235:genetics / Q000276:immunology; D006801:Humans; D010455:Peptides / Q000276:immunology; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014612:Vaccines / Q000235:genetics / Q000276:immunology; D014763:Viral Matrix Proteins / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D010455:Peptides; D014612:Vaccines; D014763:Viral Matrix Proteins