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
View: Text | PDF
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

×

Figure 1

Filarial infection selectively expands subsets of circulating effector memory CD4+ T cells.

Options: View larger image (or click on image) Download as PowerPoint
Filarial infection selectively expands subsets of circulating effector m...
(A) Schematic representation of the study summarizing the participant groups (23 filaria-infected individuals and 24 healthy donor volunteers), and gating strategies for targeting CD4+ T helper cells (B). (C) t-SNE plots generated by a FlowSOM analysis using a 27-color flow cytometry panel showing the high-dimensional diversity of CD4+ T cells among the Filaria+ (red) and Filaria– (blue) individuals. (D) Proportion of filaria-infected patients’ cells and healthy donors’ cells within each cluster; note the filaria-enriched clusters, 5 and 12. Scatter plot shows the frequency of cluster 5 (E) and cluster 12 (F) deconvoluted at the individual level by groups. (G) Histogram plot highlighting the expression level of the markers CRTH2, CCR7, CCR6, CD45RA, CCR4, and CD127 in clusters 5 (purple) and 12 (green), in comparison with naive CD4+ T cells — cluster 1 (orange). (H) Gating strategy for the conventional flow analysis for phenotypic profiling of CD4+ T cells expressing CCR4+CCR6–CRTH2–(black), CCR4+CCR6+CRTH2– (purple), CCR4+CCR6–CRTH2+ (green), and CCR4+CCR6+CRTH2+ (brown) cells. (I–L) Scatter plots show the frequency of the 4 populations described above at the individual level in their respective groups. Each dot represents a single individual, and the horizontal bars are the geometric means (GMs). Differences between the groups were considered statistically significant at P < 0.05 by unpaired Mann-Whitney U test. P values are indicated on each graph.