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The role of IFN-γ Elispot assay in HIV vaccine research

Nature Protocols volume 4, pages 461–469 (2009)Cite this article

Abstract

The interferon (IFN)-γ Elispot assay has been widely used as a general screening method for the quantification and characterization of the human immunodeficiency virus (HIV)-specific CD8+ T cell responses. However, the predictive power of this assay has been challenged due to the lack of efficacy of a recently conducted HIV vaccine phase IIb trial, despite induction of robust Elispot responses. This finding plus improvements in multiparameter flow cytometry, which has the potential advantage of simultaneously quantifying numerous parameters, raises questions regarding the future role of IFN-γ Elispot as a gateway to moving forward with clinical trials of candidate vaccines. However, the IFN-γ Elispot assay has been, unlike other techniques, evaluated and validated in several proficiency panels and is advantageous in cost-effectively detecting and mapping T-cell responses. Here we present a detailed protocol for a state-of-the-art 3-d IFN-γ Elispot assay and review further advantages and disadvantages of this method for the characterization of HIV-specific CD8+ T cell responses.

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Figure 1: Schematic drawing of the principle of the Elispot assay.
Figure 2: Anticipated results of an Elispot assay.

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Author notes

  1. hstreeck@partners.org

Authors and Affiliations

  1. Ragon Institute of HGH, MIT and Harvard, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Hendrik Streeck, Nicole Frahm & Bruce D Walker

  2. Howard Hughes Medical Institute, Chevy Chase, Maryland, USA

    Hendrik Streeck, Nicole Frahm & Bruce D Walker

Authors
  1. Hendrik Streeck
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  2. Nicole Frahm
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  3. Bruce D Walker
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Streeck, H., Frahm, N. & Walker, B. The role of IFN-γ Elispot assay in HIV vaccine research. Nat Protoc 4, 461–469 (2009). https://doi.org/10.1038/nprot.2009.7

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