Human gut microbiota–reactive DP8α Tregs prevent acute graft-versus-host disease in a CD73-dependent manner
Emmanuelle Godefroy, Patrice Chevallier, Fabienne Haspot, Caroline Vignes, Véronique Daguin, Sylvia Lambot, Margaux Verdon, Margaux De Seilhac, Valentin Letailleur, Anne Jarry, Annabelle Pédron, Thierry Guillaume, Pierre Peterlin, Alice Garnier, Marie-Anne Vibet, Maxence Mougon, Amandine Le Bourgeois, Maxime Jullien, Francine Jotereau, Frédéric Altare
Emmanuelle Godefroy, Patrice Chevallier, Fabienne Haspot, Caroline Vignes, Véronique Daguin, Sylvia Lambot, Margaux Verdon, Margaux De Seilhac, Valentin Letailleur, Anne Jarry, Annabelle Pédron, Thierry Guillaume, Pierre Peterlin, Alice Garnier, Marie-Anne Vibet, Maxence Mougon, Amandine Le Bourgeois, Maxime Jullien, Francine Jotereau, Frédéric Altare
View: Text | PDF
Research Article Immunology Transplantation

Human gut microbiota–reactive DP8α Tregs prevent acute graft-versus-host disease in a CD73-dependent manner

Abstract

Graft-versus-host disease (GvHD) is a life-threatening complication frequently occurring following allogeneic hematopoietic stem cell transplantation (allo-HSCT). Since gut microbiota and regulatory T cells (Tregs) are believed to play roles in GvHD prevention, we investigated whether DP8α Tregs, which we have previously described to harbor a T cell receptor specificity for the gut commensal Faecalibacterium prausnitzii, could protect against GvHD, thereby linking the microbiota and its effect on GvHD. We observed a decrease in CD73+ DP8α Treg frequency in allo-HSCT patients 1 month after transplantation, which was associated with acute GvHD (aGvHD) development at 1 month after transplantation, as compared with aGvHD-free patients, without being correlated to hematological disease relapse. Importantly, CD73 activity was shown to be critical for DP8α Treg suppressive function. Moreover, the frequency of host-reactive DP8α Tregs was also lower in aGvHD patients, as compared with aGvHD-free patients, which could embody a protective mechanism responsible for the maintenance of this cell subset in GvHD-free patients. We also showed that human DP8α Tregs protected mice against xenogeneic GvHD through limiting deleterious inflammation and preserving gut integrity. Altogether, these results demonstrated that human DP8α Tregs mediate aGvHD prevention in a CD73-dependent manner, likely through host reactivity, advocating for the use of these cells for the development of innovative therapeutic strategies to preclude aGvHD-related inflammation.

Authors

Emmanuelle Godefroy, Patrice Chevallier, Fabienne Haspot, Caroline Vignes, Véronique Daguin, Sylvia Lambot, Margaux Verdon, Margaux De Seilhac, Valentin Letailleur, Anne Jarry, Annabelle Pédron, Thierry Guillaume, Pierre Peterlin, Alice Garnier, Marie-Anne Vibet, Maxence Mougon, Amandine Le Bourgeois, Maxime Jullien, Francine Jotereau, Frédéric Altare

×

Figure 6

Human DP8α Tregs protect against colonic xeno-GvHD–related inflammation in vivo.

Options: View larger image (or click on image) Download as PowerPoint
Human DP8α Tregs protect against colonic xeno-GvHD–related inflammation ...
Colons were harvested at sacrifice and measured. Colonic segments were split into 2 for mechanical dissociation and subsequent flow cytometry analyses on the one hand, or histological analyses on the other hand. (A) Colon length measured at sacrifice. (B) Hematoxylin/phloxine/saffron (HPS) staining performed on 5-μm cryostat section of colon for conditioned-control mice and both treated groups. Vessels are also highlighted with arrows. (C) Crypt height was measured on 5 representative colonic regions from each mouse on sections harboring correctly oriented villi. (D) Apoptosis detection was assessed by immunohistochemistry labeling of activated caspase-3. Representative images are shown for each group of mice. (E) Vessel diameter measurements (5 measurements per animal), as highlighted in C. (F) Alcian blue staining obtained for each group of mice. Vessels are also highlighted with arrows. (G) Blue-stained areas, corresponding to secreted mucus and goblet cells producing mucus, and total crypt areas were detected using automatic structure recognition by deep learning in QuPath software. Data are expressed as the percentage of mucus area among the total crypt area from 4 representative regions from each mouse. (H) Immunostainings of human CD4 or CD8α for both groups. (I) Percentages of positive cells for CD4 or CD8α expression among the total number of cells in colonic mucosa. The percentage of positive cells was determined using the positive cell detection function of QuPath from 5 regions of interest for each mouse. Scale bars: 100 μm.