title: Effect of human bone marrow mesenchymal stromal cells on cytokine production by peripheral blood naive, memory, and effector T cells.
abstract: BACKGROUND
  The different distribution of T cells among activation/differentiation stages in immune disorders may condition the outcome of mesenchymal stromal cell (MSC)-based therapies. Indeed, the effect of MSCs in the different functional compartments of T cells is not completely elucidated.
  
  
  METHODS
  We investigated the effect of human bone marrow MSCs on naturally occurring peripheral blood functional compartments of CD4(+) and CD8(+) T cells: naive, central memory, effector memory, and effector compartments. For that, mononuclear cells (MNCs) stimulated with phorbol myristate acetate (PMA) plus ionomycin were cultured in the absence/presence of MSCs. The percentage of cells expressing tumor necrosis factor-alpha (TNF-α), interferon gamma (IFNγ), and interleukin-2 (IL-2), IL-17, IL-9, and IL-6 and the amount of cytokine produced were assessed by flow cytometry. mRNA levels of IL-4, IL-10, transforming growth factor-beta (TGF-β), and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) in purified CD4(+) and CD8(+) T cells, and phenotypic and mRNA expression changes induced by PMA + ionomycin stimulation in MSCs, were also evaluated.
  
  
  RESULTS
  MSCs induced the reduction of the percentage of CD4(+) and CD8(+) T cells producing TNF-α, IFNγ, and IL-2 in all functional compartments, except for naive IFNγ(+)CD4(+) T cells. This inhibitory effect differentially affected CD4(+) and CD8(+) T cells as well as the T-cell functional compartments; remarkably, different cytokines showed distinct patterns of inhibition regarding both the percentage of producing cells and the amount of cytokine produced. Likewise, the percentages of IL-17(+), IL-17(+)TNF-α(+), and IL-9(+) within CD4(+) and CD8(+) T cells and of IL-6(+)CD4(+) T cells were decreased in MNC-MSC co-cultures. MSCs decreased IL-10 and increased IL-4 mRNA expression in stimulated CD4(+) and CD8(+) T cells, whereas TGF-β was reduced in CD8(+) and augmented in CD4(+) T cells, with no changes for CTLA4. Finally, PMA + ionomycin stimulation did not induce significant alterations on MSCs phenotype but did increase indoleamine-2,3-dioxygenase (IDO), inducible costimulatory ligand (ICOSL), IL-1β, IL-8, and TNF-α mRNA expression.
  
  
  CONCLUSIONS
  Overall, our study showed that MSCs differentially regulate the functional compartments of CD4(+) and CD8(+) T cells, which may differentially impact their therapeutic effect in immune disorders. Furthermore, the influence of MSCs on IL-9 expression can open new possibilities for MSC-based therapy in allergic diseases.
authors.affiliation: Blood and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation, Quinta da Vinha Moura, São Martinho do Bispo, 3041-861, Coimbra, Portugal. 1979paula@gmail.com.
authors.forename: Paula
authors.initials: P
authors.lastname: Laranjeira
authors.affiliation: Blood and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation, Quinta da Vinha Moura, São Martinho do Bispo, 3041-861, Coimbra, Portugal. monia.pedrosa@ua.pt.|Signal Transduction Laboratory, Center of Cellular Biology, SACS and Departament of Biology, University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal. monia.pedrosa@ua.pt.
authors.forename: Monia
authors.initials: M
authors.lastname: Pedrosa
authors.affiliation: Blood and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation, Quinta da Vinha Moura, São Martinho do Bispo, 3041-861, Coimbra, Portugal. susanapedreiro@gmail.com.
authors.forename: Susana
authors.initials: S
authors.lastname: Pedreiro
authors.affiliation: Blood and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation, Quinta da Vinha Moura, São Martinho do Bispo, 3041-861, Coimbra, Portugal. jfgomes@ua.pt.|QOPNA, Department of Chemistry, University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal. jfgomes@ua.pt.
authors.forename: Joana
authors.initials: J
authors.lastname: Gomes
authors.affiliation: Blood and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation, Quinta da Vinha Moura, São Martinho do Bispo, 3041-861, Coimbra, Portugal. antonio.martinho@ipst.min-saude.pt.
authors.forename: Antonio
authors.initials: A
authors.lastname: Martinho
authors.affiliation: Cell2B Advanced Therapeutics SA, Biocant Park, Núcleo 04, Lote 4A, 3060-197, Cantanhede, Portugal. brigida.antunes@cell2b.com.
authors.forename: Brigida
authors.initials: B
authors.lastname: Antunes
authors.affiliation: Cell2B Advanced Therapeutics SA, Biocant Park, Núcleo 04, Lote 4A, 3060-197, Cantanhede, Portugal. catarino.tribeiro@gmail.com.
authors.forename: Tania
authors.initials: T
authors.lastname: Ribeiro
authors.affiliation: Cell2B Advanced Therapeutics SA, Biocant Park, Núcleo 04, Lote 4A, 3060-197, Cantanhede, Portugal. francisco.santos@cell2b.com.
authors.forename: Francisco
authors.initials: F
authors.lastname: Santos
authors.affiliation: Blood and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation, Quinta da Vinha Moura, São Martinho do Bispo, 3041-861, Coimbra, Portugal. helder.trindade@ipst.min-saude.pt.
authors.forename: Helder
authors.initials: H
authors.lastname: Trindade
authors.affiliation: Blood and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation, Quinta da Vinha Moura, São Martinho do Bispo, 3041-861, Coimbra, Portugal. artur.paiva@ipst.min-saude.pt.
authors.forename: Artur
authors.initials: A
authors.lastname: Paiva
mesh_terms: D000328:Adult; D001854:Bone Marrow Cells / Q000166:cytology*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism; D020381:Interleukin-17 / Q000235:genetics / Q000378:metabolism; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism; D015759:Ionomycin / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D016207:Cytokines; C508717:IL17A protein, human; D020381:Interleukin-17; D007376:Interleukin-2; D012333:RNA, Messenger; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha; D015759:Ionomycin; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate