Proteomics analysis reveals a Th17-prone cell population in presymptomatic graft-versus-host disease
Wei Li, Liangyi Liu, Aurelie Gomez, Jilu Zhang, Abdulraouf Ramadan, Qing Zhang, Sung W. Choi, Peng Zhang, Joel K. Greenson, Chen Liu, Di Jiang, Elizabeth Virts, Stephanie L. Kelich, Hong Wei Chu, Ryan Flynn, Bruce R. Blazar, Helmut Hanenberg, Samir Hanash, Sophie Paczesny
Wei Li, Liangyi Liu, Aurelie Gomez, Jilu Zhang, Abdulraouf Ramadan, Qing Zhang, Sung W. Choi, Peng Zhang, Joel K. Greenson, Chen Liu, Di Jiang, Elizabeth Virts, Stephanie L. Kelich, Hong Wei Chu, Ryan Flynn, Bruce R. Blazar, Helmut Hanenberg, Samir Hanash, Sophie Paczesny
View: Text | PDF
Research Article Immunology Transplantation

Proteomics analysis reveals a Th17-prone cell population in presymptomatic graft-versus-host disease

Abstract

Gastrointestinal graft-versus-host-disease (GI-GVHD) is a life-threatening complication occurring after allogeneic hematopoietic cell transplantation (HCT), and a blood biomarker that permits stratification of HCT patients according to their risk of developing GI-GVHD would greatly aid treatment planning. Through in-depth, large-scale proteomic profiling of presymptomatic samples, we identified a T cell population expressing both CD146, a cell adhesion molecule, and CCR5, a chemokine receptor that is upregulated as early as 14 days after transplantation in patients who develop GI-GVHD. The CD4+CD146+CCR5+ T cell population is Th17 prone and increased by ICOS stimulation. shRNA knockdown of CD146 in T cells reduced their transmigration through endothelial cells, and maraviroc, a CCR5 inhibitor, reduced chemotaxis of the CD4+CD146+CCR5+ T cell population toward CCL14. Mice that received CD146 shRNA–transduced human T cells did not lose weight, showed better survival, and had fewer CD4+CD146+CCR5+ T cells and less pathogenic Th17 infiltration in the intestine, even compared with mice receiving maraviroc with control shRNA–transduced human T cells. Furthermore, the frequency of CD4+CD146+CCR5+ Tregs was increased in GI-GVHD patients, and these cells showed increased plasticity toward Th17 upon ICOS stimulation. Our findings can be applied to early risk stratification, as well as specific preventative therapeutic strategies following HCT.

Authors

Wei Li, Liangyi Liu, Aurelie Gomez, Jilu Zhang, Abdulraouf Ramadan, Qing Zhang, Sung W. Choi, Peng Zhang, Joel K. Greenson, Chen Liu, Di Jiang, Elizabeth Virts, Stephanie L. Kelich, Hong Wei Chu, Ryan Flynn, Bruce R. Blazar, Helmut Hanenberg, Samir Hanash, Sophie Paczesny

×

Figure 3

Endothelial CD146 expression in GI-GVHD colonic biopsies, transendothelial migration of CD4+ T cell subsets and with CD146 knockdown, and chemotaxis of CD4+CD146+CCR5+ T cells.

Options: View larger image (or click on image) Download as PowerPoint
Endothelial CD146 expression in GI-GVHD colonic biopsies, transendotheli...
(A) Immunohistochemical analysis of CD146 expression in colonic biopsies taken at onset of symptoms from non-GVHD enteritis patients and GI-GVHD patients (magnification ×200). Dot plot showing mean ± SEM values for CD146+ vessel counts ×10, 2-tailed Student’s t test from non-GVHD enteritis patients (n = 10) and GI-GVHD patients (n = 18). (B–E) Transendothelial migration of CD146– and CD146+ T cells sorted from PB cells, Th1 and Th17 cells, Th17 cells with CD146 knockdown via CD146 shRNA1, or CD146 shRNA2. Representative flow cytometric histograms showing the efficiency of CD146 knockdown. Isotype control staining (gray) and CD146 staining of cells with the control shRNA (red) and CD146 shRNA (blue). Dot plots show mean ± SEM values for percentage of transmigrated CD4+ T cells (n = 6 for B, n = 5 for C, n = 5 for D, and n = 8 for E), 2-tailed Student’s t test. (F) CCR5-mediated chemotaxis of CD4+CD146+CCR5+ T cells toward CCL14 or CCL14 and CCL5. The sorted double-positive cells were pretreated with maraviroc (MV) or vehicle control. Data are presented as mean ± SEM values (n = 3), paired t test.