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Background  The immunomodulatory effects of granulocyte colony-stimulating factor (G-CSF) on T cells result in a low incidence of acute graft-versus-host disease (aGVHD) in G-CSF-mobilized allogeneic peripheral blood stem cell transplantation (G-PBSCT). However, the exact mechanism remains unclear. Regulatory γδ T cells (γδTregs), characterized by the presence of TCRγδ and Foxp3, have aroused great concern in the maintenance of immune tolerance. We hypothesized that γδTregs might involve in the immunomodulatory effects of G-CSF mobilization.    Methods  The expression and immunomodulatory function of γδTreg subsets in peripheral blood of donors before and after G-CSF treatment in vivo and in vitro were evaluated by flow cytometry and CFSE assays. To investigate the effects of γδTregs on aGVHD, the association between γδTreg subsets in grafts and aGVHD in recipients was estimated.    Results  The proportions of Vδ1Tregs, CD27 + Vδ1Tregs and CD25 + Vδ1Tregs were significantly increased in peripheral blood after G-CSF treatment in vivo. γδTregs could be generated in vitro by stimulating with anti-TCRγδ in the presence of G-CSF. The immune phenotype, proliferation suppression function, and cytokine secretion of G-CSF-induced γδTregs were similar to that of transforming growth factor-β (TGF-β)-induced γδTregs. The clinical data demonstrated that the proportion of CD27 + Vδ1Tregs in grafts was significantly lower in the patients who experienced aGVHD than in those who did not develop aGVHD ( P  = 0.028), and the proportions of other γδTreg subsets in grafts did not differ significantly between the two groups. The best cutoff value for CD27 + Vδ1Treg proportion in grafts in prediction of aGVHD was 0.33%, with an area under the curve value of 0.725 ( P  = 0.043). Eight patients (26.7%) were classified as the low-CD27 + Vδ1Treg group (< 0.33%), and 22 patients (73.3%) as the high-CD27 + Vδ1Treg group (≥ 0.33%). The incidence of aGVHD was higher in the low-CD27 + Vδ1Treg group than in the high-CD27 + Vδ1Treg group (75.0% versus 22.7%,  P  = 0.028).    Conclusions  G-CSF could induce the generation of γδTregs in vivo and in vitro, and γδTregs might participate in aGVHD regulation in G-PBSCT.

journal of translational medicine

Regulatory γδ T cells induced by G-CSF participate in acute graft-versus-host disease regulation in G-CSF-mobilized allogeneic peripheral blood stem cell transplantation