Polyfunctional and IFN-γ monofunctional human CD4+ T cell populations are molecularly distinct
Julie G. Burel, Simon H. Apte, Penny L. Groves, James S. McCarthy, Denise L. Doolan
Julie G. Burel, Simon H. Apte, Penny L. Groves, James S. McCarthy, Denise L. Doolan
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Research Article Cell biology

Polyfunctional and IFN-γ monofunctional human CD4+ T cell populations are molecularly distinct

Abstract

Pathogen-specific polyfunctional T cell responses have been associated with favorable clinical outcomes, but it is not known whether molecular differences exist between polyfunctional and monofunctional cytokine-producing T cells. Here, we report that polyfunctional CD4+ T cells induced during Plasmodium falciparum (P. falciparum) blood-stage infection in humans have a unique transcriptomic profile compared with IFN-γ monofunctional CD4+ T cells and, thus, are molecularly distinct. The 14-gene signature revealed in P. falciparum–reactive polyfunctional T cells is associated with cytokine signaling and lymphocyte chemotaxis, and systems biology analysis identified IL-27 as an upstream regulator of the polyfunctional gene signature. Importantly, the polyfunctional gene signature is largely conserved in Influenza-reactive polyfunctional CD4+ T cells, suggesting that polyfunctional T cells have core characteristics independent of pathogen specificity. This study provides the first evidence to our knowledge that consistent molecular differences exist between polyfunctional and monofunctional CD4+ T cells.

Authors

Julie G. Burel, Simon H. Apte, Penny L. Groves, James S. McCarthy, Denise L. Doolan

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Figure 5

Differential gene expression profile between IFN-γ single positive and IFN-γ/IL-2/TNF-α triple positive CD4+ T cells with P. falciparum or Influenza reactivity determined by multiplex qPCR.

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Differential gene expression profile between IFN-γ single positive and I...
Fold change in gene expression between IFN-γ single positive and IFN-γ/IL-2/TNF-α triple positive CD4+ T cells that were reactive to P. falciparum (left) or Influenza (right). P. falciparum–reactive CD4+ T cells were isolated after in vitro stimulation, with Pf prbc parasite antigen extract overnight of whole blood samples collected from 5 volunteers at 4 weeks following experimental infection with P. falciparum blood stage. Influenza-reactive CD4+ T cells were isolated after Influenza peptide stimulation of whole blood collected from 5 healthy volunteers at 5 weeks following immunization with seasonal Influenza trivalent vaccine. Cytokine-producing CD4+ T cells were isolated using a 3-color polyfunctional cytokine secretion assay, and the expression of 37 genes selected from the Affymetrix microarray analysis was determined by multiplex qPCR (Fluidigm Biomark HD system). Fold change in gene expression between IFN-γ single positive and IFN-γ/IL-2/TNF-α triple positive CD4+ T cells was determined by the ddCt method. (A) Genes upregulated in both P. falciparum–reactive and Influenza-reactive IFN-γ/IL-2/TNF-α triple positive CD4+ T cells. (B and C) Genes upregulated only in P. falciparum–reactive or Influenza-reactive IFN-γ/IL-2/TNF-α triple positive CD4+ T cells, respectively. (D) Genes with no consistent gene expression across individuals for both antigens.