title: G-CSF-Induced Suppressor IL-10+ Neutrophils Promote Regulatory T Cells That Inhibit Graft-Versus-Host Disease in a Long-Lasting and Specific Way.
abstract: Acute graft-versus-host disease (aGVHD) is the main complication of allogeneic hematopoietic stem cell transplantation, and many efforts have been made to overcome this important limitation. We showed previously that G-CSF treatment generates low-density splenic granulocytes that inhibit experimental aGVHD. In this article, we show that aGVHD protection relies on incoming IL-10+neutrophils from G-CSF-treated donor spleen (G-Neutrophils). These G-Neutrophils have high phagocytic capacity, high peroxide production, low myeloperoxidase activity, and low cytoplasmic granule content, which accounts for their low density. Furthermore, they have low expression of MHC class II, costimulatory molecules, and low arginase1 expression. Also, they have low IFN-γ, IL-17F, IL-2, and IL-12 levels, with increased IL-10 production and NO synthase 2 expression. These features are in accordance with the modulatory capacity of G-Neutrophils on regulatory T cell (Treg) generation. In vivo, CD25+Treg depletion shortly after transplantation with splenic cells from G-CSF-treated donors blocks suppression of aGVHD, suggesting Treg involvement in the protection induced by the G-Neutrophils. The immunocompetence and specificity of the semiallogeneic T cells, long-term after the bone marrow transplant using G-Neutrophils, were confirmed by third-party skin graft rejection; importantly, a graft-versus-leukemia assay showed that T cell activity was maintained, and all of the leukemic cells were eliminated. We conclude that G-CSF treatment generates a population of activated and suppressive G-Neutrophils that reduces aGVHD in an IL-10- and Treg-dependent manner, while maintaining immunocompetence and the graft versus leukemia effect.
authors.affiliation: Departamento de Imunologia, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-590, Brazil.|Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro 20231-050, Brazil.|Laboratório de Pesquisa Sobre o Timo, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro 21040-360, Brazil.
authors.forename: Suelen Martins
authors.identifier: 0000-0003-1146-4647
authors.initials: SM
authors.lastname: Perobelli
authors.affiliation: Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro 20231-050, Brazil.
authors.forename: Ana Carolina Terra
authors.initials: AC
authors.lastname: Mercadante
authors.affiliation: Departamento de Imunologia, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-590, Brazil.|Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro 20231-050, Brazil.|Laboratório de Pesquisa Sobre o Timo, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro 21040-360, Brazil.
authors.forename: Rômulo Gonçalves
authors.initials: RG
authors.lastname: Galvani
authors.affiliation: Departamento de Imunologia, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-590, Brazil.|Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro 20231-050, Brazil.|Laboratório de Pesquisa Sobre o Timo, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro 21040-360, Brazil.
authors.forename: Triciana
authors.initials: T
authors.lastname: Gonçalves-Silva
authors.affiliation: Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro 20231-050, Brazil.
authors.forename: Ana Paula Gregório
authors.identifier: 0000-0001-5969-1115
authors.initials: AP
authors.lastname: Alves
authors.affiliation: Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-590, Brazil.|Departamento de Microbiologia, FIOCRUZ Pernambuco, Pernambuco 50670-420, Brazil.
authors.forename: Antonio
authors.identifier: 0000-0001-6629-568X
authors.initials: A
authors.lastname: Pereira-Neves
authors.affiliation: Universidade do Grande, Rio de Janeiro 25071-202, Brazil.|Instituto Nacional de Biologia Estrutural e Bioimagem da Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-590, Brazil; and.
authors.forename: Marlene
authors.identifier: 0000-0002-4957-8538
authors.initials: M
authors.lastname: Benchimol
authors.affiliation: Departamento de Imunologia, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-590, Brazil.
authors.forename: Alberto
authors.initials: A
authors.lastname: Nóbrega
authors.affiliation: Departamento de Imunologia, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-590, Brazil; adriana.bonomo@fiocruz.br acbonomo@gmail.com.|Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro 20231-050, Brazil.|Laboratório de Pesquisa Sobre o Timo, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro 21040-360, Brazil.|Programa FIOCancer, Vice Presidência de Pesquisa e Laboratórios de Referência/FIOCRUZ, Rio de Janeiro 21040-360, Brazil.
authors.forename: Adriana
authors.identifier: 0000-0002-3938-9514
authors.initials: A
authors.lastname: Bonomo
mesh_terms: D000208:Acute Disease; D000818:Animals; D002478:Cells, Cultured; D006086:Graft vs Host Disease / Q000276:immunology* / Q000517:prevention & control; D016179:Granulocyte Colony-Stimulating Factor / Q000276:immunology / Q000378:metabolism*; D018380:Hematopoietic Stem Cell Transplantation; D007108:Immune Tolerance; D016753:Interleukin-10 / Q000378:metabolism; D008212:Lymphocyte Depletion; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology*; D010587:Phagocytosis; D017382:Reactive Oxygen Species / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D014184:Transplantation, Homologous
chemical_list: D017382:Reactive Oxygen Species; D016753:Interleukin-10; D016179:Granulocyte Colony-Stimulating Factor