Skip to main content
Journal cover image

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

Publication ,  Journal Article
Currier, JR; Kuta, EG; Turk, E; Earhart, LB; Loomis-Price, L; Janetzki, S; Ferrari, G; Birx, DL; Cox, JH
Published in: J Immunol Methods
February 1, 2002

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.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

J Immunol Methods

DOI

ISSN

0022-1759

Publication Date

February 1, 2002

Volume

260

Issue

1-2

Start / End Page

157 / 172

Location

Netherlands

Related Subject Headings

  • Quality Control
  • Lymphocyte Activation
  • Immunosorbent Techniques
  • Immunology
  • Humans
  • Histocompatibility Antigens Class I
  • HIV Antigens
  • CD8-Positive T-Lymphocytes
  • AIDS Vaccines
  • 3204 Immunology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Currier, J. R., Kuta, E. G., Turk, E., Earhart, L. B., Loomis-Price, L., Janetzki, S., … Cox, J. H. (2002). A panel of MHC class I restricted viral peptides for use as a quality control for vaccine trial ELISPOT assays. J Immunol Methods, 260(1–2), 157–172. https://doi.org/10.1016/s0022-1759(01)00535-x
Currier, Jeffrey R., Ellen G. Kuta, Ellen Turk, Lyndsay B. Earhart, Larry Loomis-Price, Sylvia Janetzki, Guido Ferrari, Deborah L. Birx, and Josephine H. Cox. “A panel of MHC class I restricted viral peptides for use as a quality control for vaccine trial ELISPOT assays.J Immunol Methods 260, no. 1–2 (February 1, 2002): 157–72. https://doi.org/10.1016/s0022-1759(01)00535-x.
Currier JR, Kuta EG, Turk E, Earhart LB, Loomis-Price L, Janetzki S, et al. 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.
Currier, Jeffrey R., et al. “A panel of MHC class I restricted viral peptides for use as a quality control for vaccine trial ELISPOT assays.J Immunol Methods, vol. 260, no. 1–2, Feb. 2002, pp. 157–72. Pubmed, doi:10.1016/s0022-1759(01)00535-x.
Currier JR, Kuta EG, Turk E, Earhart LB, Loomis-Price L, Janetzki S, Ferrari G, Birx DL, Cox JH. 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):157–172.
Journal cover image

Published In

J Immunol Methods

DOI

ISSN

0022-1759

Publication Date

February 1, 2002

Volume

260

Issue

1-2

Start / End Page

157 / 172

Location

Netherlands

Related Subject Headings

  • Quality Control
  • Lymphocyte Activation
  • Immunosorbent Techniques
  • Immunology
  • Humans
  • Histocompatibility Antigens Class I
  • HIV Antigens
  • CD8-Positive T-Lymphocytes
  • AIDS Vaccines
  • 3204 Immunology