A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-γ HIV-specific immune responses

Salix Boulet, Michel L. Ndongala, Yoav Peretz, Marie Pierre Boisvert, Mohamed Rachid Boulassel, Cecile Tremblay, Jean Pierre Routy, Rafick P. Sekaly, Nicole F. Bernard

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The single color IFN-γ ELISPOT assay has become a standard for assessing HIV-specific immune responses in HIV-infected subjects. However, recent data suggests that single cytokine detection for immune monitoring of HIV-infected individuals may not be sufficient to fully describe virus-specific immune responses. Here, we have designed and validated a dual color ELISPOT assay capable of detecting both IL-2 and IFN-γ secreting cells simultaneously in response to HIV antigens. We found that a cell input number of 200,000 cells/well provided a good balance between limited availability of cells due to blood volume restrictions and ability to detect all cytokine secretion patterns. The simultaneous detection of IL-2 and IFN-γ resulted in a decreased magnitude of IFN-γ but not IL-2 responses. Measures of intra- and inter-assay variability for the dual color ELISPOT assay were comparable to that seen for single cytokine ELISPOT assay with coefficients of variation below 20% for IL-2, IFN-γ and dual secretion. Although CD8+ T cells mediated most HIV-specific responses in infected subjects, CD4+ T cells mediated responses to HIV were also detected. Features of this assay such as high throughput, cell number requirement and cytokine choice should make this assay a valuable tool for screening for HIV-specific immune responses in several clinically relevant settings.

Original languageEnglish (US)
Pages (from-to)18-29
Number of pages12
JournalJournal of Immunological Methods
Volume320
Issue number1-2
DOIs
StatePublished - Mar 30 2007
Externally publishedYes

Fingerprint

Enzyme-Linked Immunospot Assay
Interleukin-2
Color
HIV
Cytokines
Cell Count
HIV Antigens
T-Lymphocytes
Immunologic Monitoring
Blood Volume
Viruses

Keywords

  • CD4+
  • CD8+
  • Cellular immune responses
  • ELISPOT
  • HIV
  • IL-2
  • Interferon-gamma
  • T cells

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Boulet, S., Ndongala, M. L., Peretz, Y., Boisvert, M. P., Boulassel, M. R., Tremblay, C., ... Bernard, N. F. (2007). A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-γ HIV-specific immune responses. Journal of Immunological Methods, 320(1-2), 18-29. https://doi.org/10.1016/j.jim.2006.11.010

A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-γ HIV-specific immune responses. / Boulet, Salix; Ndongala, Michel L.; Peretz, Yoav; Boisvert, Marie Pierre; Boulassel, Mohamed Rachid; Tremblay, Cecile; Routy, Jean Pierre; Sekaly, Rafick P.; Bernard, Nicole F.

In: Journal of Immunological Methods, Vol. 320, No. 1-2, 30.03.2007, p. 18-29.

Research output: Contribution to journalArticle

Boulet, S, Ndongala, ML, Peretz, Y, Boisvert, MP, Boulassel, MR, Tremblay, C, Routy, JP, Sekaly, RP & Bernard, NF 2007, 'A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-γ HIV-specific immune responses', Journal of Immunological Methods, vol. 320, no. 1-2, pp. 18-29. https://doi.org/10.1016/j.jim.2006.11.010
Boulet, Salix ; Ndongala, Michel L. ; Peretz, Yoav ; Boisvert, Marie Pierre ; Boulassel, Mohamed Rachid ; Tremblay, Cecile ; Routy, Jean Pierre ; Sekaly, Rafick P. ; Bernard, Nicole F. / A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-γ HIV-specific immune responses. In: Journal of Immunological Methods. 2007 ; Vol. 320, No. 1-2. pp. 18-29.
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