Single-cell immunophenotyping identifies CD8+GZMK+IFNG+ T cells as a key immune population in cutaneous Lyme disease
Edel Aron, Hailong Meng, Alexia A. Belperron, Paraskevas Filippidis, Kenneth R. Dardick, Steven H. Kleinstein, Linda K. Bockenstedt
Edel Aron, Hailong Meng, Alexia A. Belperron, Paraskevas Filippidis, Kenneth R. Dardick, Steven H. Kleinstein, Linda K. Bockenstedt
View: Text | PDF
Research Article Immunology Infectious disease

Single-cell immunophenotyping identifies CD8+GZMK+IFNG+ T cells as a key immune population in cutaneous Lyme disease

Abstract

The skin lesion erythema migrans (EM) is the first clinical sign of Lyme disease, an infection due to the tick-transmitted bacterium Borrelia burgdorferi (Bb). Previously, we used scRNA-Seq to characterize the cutaneous immune response in the EM lesion, focusing on B cells. Here, with an expanded sample size, we profiled T cell responses in EM lesions compared to autologous uninvolved skin. In addition to CD4+ T cell subsets known to be abundant in the EM lesion, we identified clonally expanded CD8+GZMK+IFNG+ T cells that comprised cells with high or intermediate IFNG expression. These cells exhibited significant differential expression of IFN-regulated genes and included subsets with low cytotoxic gene expression, suggesting an inflammatory potential that may contribute to early defense against Bb within the EM lesion. In addition, we found that endothelial cells, fibroblasts, and pericytes were the main producers of key T cell–recruiting chemokines. These studies using single-cell transcriptomics with adaptive immune receptor sequencing provide a comprehensive interrogation of the cutaneous T cell response to Bb infection and insight into the orchestration of the skin barrier defense to this vector-borne pathogen.

Authors

Edel Aron, Hailong Meng, Alexia A. Belperron, Paraskevas Filippidis, Kenneth R. Dardick, Steven H. Kleinstein, Linda K. Bockenstedt

×

Figure 2

Clustering and annotation of the skin T cells.

Options: View larger image (or click on image) Download as PowerPoint
Clustering and annotation of the skin T cells. (A) UMAP representing clu...
(A) UMAP representing clustering of the subclustered T cells from the combined control and EM skin data. At a resolution of 1.5, there were 26 clusters formed. (B) UMAP of skin T cell types annotated using canonical marker genes. (C and D) Proportions of cell types within the control and EM samples, respectively, arranged in decreasing order by percentage. Proportions were averaged by participant for each cell type. Colors correspond to the cell types in B. (E) Comparison of the frequencies of each cell type between control and EM samples. Cell frequencies are relative to all skin cells, not just the total T cells. Points represent results from individual participants and the dashed lines connect paired samples. The solid black horizontal lines within each box plot indicate the median and the blue or orange lines indicate the mean for each sample type and cell type across participants. Each box plot is bounded by the 25th and 75th percentiles (bottom and top lines respectively) of the data, with whiskers extending to the minimum and maximum values and additional points representing outliers. Gray backgrounds indicate the cell types that did not have significant increases in EM compared to control samples. Significance was calculated using a paired Wilcoxon’s signed-rank test (**P < 0.01, *P < 0.05).