Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2017 Mar 6;8:103. doi: 10.3389/fimmu.2017.00103

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2017 Duurland, Brown, O’Shaughnessy and Wedderburn.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

Shared canonical signature between CD161⁺ conventional T cells (Tconv) and CD161⁺ regulatory T cells (Treg) reveals a T helper (Th) phenotype of CD161⁺ T cells. Healthy adult peripheral blood mononuclear cells (PBMC) (n = 4) were sorted into CD4⁺CD127⁺CD25⁻CD161⁺ (CD161⁺ Tconv) or CD161⁻ (CD161⁻ Tconv), and CD4⁺CD127^lowCD25^hiCD161⁺ (CD161⁺ Treg) or CD161⁻ (CD161⁻ Treg) for RNAseq analysis. (A) Principal component analysis (PCA) of data from all samples before applying filters to assess differentially expressed (DE) genes. One CD161⁺ Treg sample was excluded from analysis due to low cDNA yield. (B) Heat maps of DE genes comparing CD161⁺ and CD161⁻ Tconv (CD161⁺ Tconv signature) (1626 DE genes, left), and CD161⁺ and CD161⁻ Treg (CD161⁺ Treg signature) (826 DE genes, right). (C) Venn diagram representing overlap in transcriptional signatures of CD161⁺ Tconv and CD161⁺ Treg. (D) Canonical pathways altered in both CD161⁺ Tconv (blue bars) and CD161⁺ Treg (red bars) signatures identified using ingenuity pathway analysis with −log P-value >2 (equals P-value <0.01). (E) DE genes associated with Th1 and Th17 in CD161⁺ Tconv and CD161⁺ Treg compared to CD161⁻ counterparts. (F) Analysis of CCR5 expression; normalized expression values (left, from RNAseq) and percentage CCR5⁺ cells (right, flow cytometry) within CD161⁻ (○) and CD161⁺ () Tconv and Treg in healthy adult (n = 10) and child (n = 4) PBMC. (G,H) Analysis of CXCR3 (G) and IL18R (H) expression; normalized expression values (left, from RNAseq) and protein expression (right, flow cytometry) within CD161⁻ (○) and CD161⁺ () Tconv and Treg in healthy adult (n = 9–15) and child (n = 4–5) PBMC. (I,J) PBMC (I) or sorted CD161⁺ and CD161⁻ Tconv (J) from healthy adults were cultured in the presence of 50 ng/ml IL-18 and IL-12 for 24 h with Brefeldin A for last 4 h of culture. (I) Representative plots and summary graphs showing percentage IFNγ⁺ within CD161⁻ (○) and CD161⁺ () Tconv and Treg (n = 8). (J) Representative plots showing IFNγ⁺ cells within sorted CD161⁺ and CD161⁻ Tconv from 2 healthy adult controls. Statistical significance: *P < 0.05, **P < 0.01, ***P < 0.001.

Shared canonical signature between CD161⁺ conventional T cells (Tconv) and CD161⁺ regulatory T cells (Treg) reveals a T helper (Th) phenotype of CD161⁺ T cells. Healthy adult peripheral blood mononuclear cells (PBMC) (n = 4) were sorted into CD4⁺CD127⁺CD25⁻CD161⁺ (CD161⁺ Tconv) or CD161⁻ (CD161⁻ Tconv), and CD4⁺CD127^lowCD25^hiCD161⁺ (CD161⁺ Treg) or CD161⁻ (CD161⁻ Treg) for RNAseq analysis. (A) Principal component analysis (PCA) of data from all samples before applying filters to assess differentially expressed (DE) genes. One CD161⁺ Treg sample was excluded from analysis due to low cDNA yield. (B) Heat maps of DE genes comparing CD161⁺ and CD161⁻ Tconv (CD161⁺ Tconv signature) (1626 DE genes, left), and CD161⁺ and CD161⁻ Treg (CD161⁺ Treg signature) (826 DE genes, right). (C) Venn diagram representing overlap in transcriptional signatures of CD161⁺ Tconv and CD161⁺ Treg. (D) Canonical pathways altered in both CD161⁺ Tconv (blue bars) and CD161⁺ Treg (red bars) signatures identified using ingenuity pathway analysis with −log P-value >2 (equals P-value <0.01). (E) DE genes associated with Th1 and Th17 in CD161⁺ Tconv and CD161⁺ Treg compared to CD161⁻ counterparts. (F) Analysis of CCR5 expression; normalized expression values (left, from RNAseq) and percentage CCR5⁺ cells (right, flow cytometry) within CD161⁻ (○) and CD161⁺ () Tconv and Treg in healthy adult (n = 10) and child (n = 4) PBMC. (G,H) Analysis of CXCR3 (G) and IL18R (H) expression; normalized expression values (left, from RNAseq) and protein expression (right, flow cytometry) within CD161⁻ (○) and CD161⁺ () Tconv and Treg in healthy adult (n = 9–15) and child (n = 4–5) PBMC. (I,J) PBMC (I) or sorted CD161⁺ and CD161⁻ Tconv (J) from healthy adults were cultured in the presence of 50 ng/ml IL-18 and IL-12 for 24 h with Brefeldin A for last 4 h of culture. (I) Representative plots and summary graphs showing percentage IFNγ⁺ within CD161⁻ (○) and CD161⁺ () Tconv and Treg (n = 8). (J) Representative plots showing IFNγ⁺ cells within sorted CD161⁺ and CD161⁻ Tconv from 2 healthy adult controls. Statistical significance: *P < 0.05, **P < 0.01, ***P < 0.001.