A panel of MHC class I restricted viral peptides for use as a quality control for vaccine trial ELISPOT assays

J Immunol Methods. 2002 Feb 1;260(1-2):157-72. doi: 10.1016/s0022-1759(01)00535-x.


Vaccines in general and HIV vaccines in particular are focusing ever more on the induction of cellular immunity specifically the generation of cytotoxic T cells (CTL). As progress is made towards developing a safe and effective HIV vaccine, there is a need for a robust, sensitive and reproducible assay to evaluate vaccine-induced cellular immunogenicity in Phase II/III trials. The enzyme-linked immunospot (ELISPOT) assay fits these criteria and is a technology that is readily transferable and amenable to high-through-put screening. There is a need for reagents that can be used as positive controls and for optimizing and standardizing the assay. We selected a panel of 23 8-11 mer Influenza virus (Flu), Cytomegalovirus (CMV) and Epstein Barr virus (EBV) epitopes recognized by CD8 positive T cells and presented by 11 class I HLA-A and HLA-B alleles whose cumulative frequencies represent >100% of Caucasian individuals. We examined interferon-gamma (IFN-gamma) secretion in peripheral blood mononuclear cells (PBMC) incubated with the peptides using a modified ELISPOT assay. IFN-gamma secretion was detected in 15/17 (88%) HIV-1 seronegative and 14/20 (70%) HIV-1 seropositive individuals. Release of IFN-gamma in response to the pool of peptides was CD8+ T cell mediated and HLA restricted. In vitro stimulation of PBMC with individual peptides or the pool of peptides led to the expansion of T cells capable of killing target cells expressing the appropriate viral antigen in a CTL assay. The size, shape and appearance of the spots produced using this peptide panel provided a standard for the establishment of acceptance criteria of spots for the evaluation of ELISPOT plates using an automated reader system.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • AIDS Vaccines*
  • CD8-Positive T-Lymphocytes / immunology
  • HIV Antigens / analysis*
  • HIV Antigens / immunology
  • Histocompatibility Antigens Class I / immunology
  • Humans
  • Immunosorbent Techniques*
  • Lymphocyte Activation
  • Quality Control


  • AIDS Vaccines
  • HIV Antigens
  • Histocompatibility Antigens Class I