Brice, Gary T., Graber, Norma L., Hoffman, Stephen L., and Doolan, Denise L. (2001) Expression of the chemokine MIG is a sensitive and predictive marker for antigen-specific, genetically restricted IFN-γ production and IFN-γ-secreting cells. Journal of Immunological Methods, 257 (1-2). pp. 55-69.

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Abstract

The evaluation of antigen-specific immune responses is critical for understanding the mechanisms of immune protection and for establishing the efficacy of candidate vaccines. Here, we describe a novel assay for IFN-γ activity which is based on the flow cytometric detection of the chemokine, monokine induced by gamma interferon (MIG) as a sensitive and predictive measure of IFN-γ-mediated effector function, and a surrogate marker for IFN-γ-producing cells. Upregulation of MIG expression was demonstrated following in vitro activation of peripheral blood mononuclear cells (PBMCs) with defined CD8⁺ T-cell epitopes derived from influenza virus, cytomegalovirus (CMV), or Epstein-Barr virus (EBV) and was antigen-specific, genetically restricted and dependent on both CD8⁺ T cells and IFN-γ. Furthermore, antigen-specific MIG expression was also demonstrated with Plasmodium falciparum circumsporozoite protein (CSP) peptides, using PBMCs from volunteers immunized with irradiated P. falciparum sporozoites. In multiple parallel experiments, the MIG assay was compared to conventional IFN-γ ELISPOT, IFN-γ ELISA, MIG ELISA and intracellular cytokine staining assays. The level of MIG expression was shown to be directly associated with the number of IFN-γ spot-forming cells (SFCs) detected by ELISPOT (r²=0.94). Moreover, in all instances where cultures were considered positive by ELISPOT, a higher stimulation index was noted with the MIG assay as compared with the ELISPOT assay (on average at least threefold higher) and, in some cases, responses as detected by the MIG assay were significant, but the corresponding response as measured by ELISPOT was not significant. Finally, the flow-based MIG assay offers a number of practical and technical advantages over the ELISPOT assay. Our data validate this novel method for the detection of low as well as high levels of antigen-specific and genetically restricted IFN-γ activity.

Item ID:	42764
Item Type:	Article (Research - C1)
ISSN:	1872-7905
Keywords:	chemokines; cytokines; CD8+ T lymphocytes; IFN-γ
Funders:	Naval Medical Research Centre (NMRC)
Projects and Grants:	NMRC 61102A.S13.F.A0009, NMRC 62787A.870.F.A0010
Date Deposited:	21 Mar 2016 04:44
FoR Codes:	11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110799 Immunology not elsewhere classified @ 100%
SEO Codes:	92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920109 Infectious Diseases @ 100%
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