Single-cell molecular signature of pathogenic T helper subsets in type 2–associated disorders in humans
Pedro H. Gazzinelli-Guimaraes, Brittany Dulek, Phillip Swanson, Justin Lack, Mario Roederer, Thomas B. Nutman
Pedro H. Gazzinelli-Guimaraes, Brittany Dulek, Phillip Swanson, Justin Lack, Mario Roederer, Thomas B. Nutman
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Research Article Immunology Infectious disease

Single-cell molecular signature of pathogenic T helper subsets in type 2–associated disorders in humans

Abstract

To unravel the heterogeneity and molecular signature of effector memory Th2 cells (Tem2), we analyzed 23 individuals’ PBMCs of filaria-infected (Filaria+) and 24 healthy volunteers (Filaria–), with or without coincident house dust mite (HDM) allergic sensitization. Flow cytometry revealed 3 CD4+ Tem subsets — CCR4+CCR6+CRTH2– Tem17, CCR4+CCR6-CRTH2+ Tem2, and CCR6+CCR4+CRTH2+ Tem17.2 — markedly enriched in Filaria+ individuals. These subsets were sorted and analyzed by multiomic single-cell RNA immunoprofiling. SingleR-annotated Th2 cells from Tem2 and Tem17.2 cell subsets had features of pathogenic Th2 effector cells based on their transcriptional signatures, with downregulated CD27 and elevated expression levels of ITGA4, IL17RB, HPGDS, KLRB1, PTGDR2, IL9R, IL4, IL5, and IL13 genes. When the Filaria+ individuals were subdivided based on their allergic status, Tem2 cells in HDM+Filaria+ individuals showed an overall reduction in TCR diversity, suggesting the occurrence of antigen-driven clonal expansion. Moreover, HDM+Filaria+ individuals showed not only an expansion in the frequency of both Tem2 and Tem17.2 cell subsets, but also a change in their molecular program by overexpressing GATA3, IL17RB, CLRF2, and KLRB1, as well as increased antigen-induced IL-4, IL-5, and IL-13 production, suggesting that aeroallergens reshape the transcriptional and functional programming of Th2 cell subsets in human filarial infection toward a pathogenic immunophenotype.

Authors

Pedro H. Gazzinelli-Guimaraes, Brittany Dulek, Phillip Swanson, Justin Lack, Mario Roederer, Thomas B. Nutman

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Figure 2

Functional diversity of CD4+ T cell subsets demonstrate type 2 cytokine signatures in filarial infection.

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Functional diversity of CD4+ T cell subsets demonstrate type 2 cytokine ...
(A) Heatmap showing the net production of IL-4, IL-5, IL-6, IL-9, IL-10, and IL-13 levels (pg/mL transformed in z score) in the culture supernatant after in vitro stimulation with BMA (10 μg/mL). Each column represents a single individual. The scatter plot shows the frequency of CCR4+CCR6+CRTH2– (B), CCR4+CCR6–CRTH2+ (C), and CCR4+CCR6+CRTH2+ (D) CD4+ Tem cells from PBMCs of filaria-uninfected and filaria-infected patients producing IL-4, IL-5, or IL-13. Each dot represents a single individual, and the horizontal bars are the GMs. Differences between the groups were considered statistically significant at P < 0.05 by the Mann-Whitney U test. P values are indicated on each graph. The frequency of IL-4– (E), IL-5– (H), and IL-13–producing (K) CD4+ Tem cells by intracellular staining following 12-hour stimulation with PMA/ionomycin and brefeldin A. The proportions of each Th cell subset, CCR4+CCR6+CRTH2– (purple), CCR4+CCR6–CRTH2+ (green), and CCR4+CCR6+CRTH2+ (brown), and other cells (gray) from healthy donors (F, I, and L) and filaria-infected patients (G, J, and M).