title: A simple method for identifying bone marrow mesenchymal stromal cells with a high immunosuppressive potential.
abstract: OBJECTIVE
  The beneficial activity of mesenchymal stromal cells (MSC) in allogeneic hematopietic stem cell transplantation requires correct use in terms of cell dose and timing of infusion and the identification of biomarkers for selection. The immunosuppressive bone marrow (BM)-derived MSC (BM-MSC) functions have been associated with the production of soluble HLA-G molecules (sHLA-G) via interleukin (IL)-10. We have established a reliable method for evaluating BM-MSC HLA-G expression without the influence of peripheral blood mononuclear cells (PBMC).
  
  
  METHODS
  Thirteen BM-MSC from donors were activated with recombinant IL-10 or co-cultured with 10 different phytohemagglutinin (PHA)-treated PBMC (PHA-PBMC). Membrane-bound and sHLA-G expression was evaluated by flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively; lymphoproliferation was measured by (methyl-(3)H)thymidine.
  
  
  RESULTS
  The results demonstrated the ability of IL-10 to stimulate both membrane-bound and sHLA-G production by BM-MSC. The levels of HLA-G expression induced by IL-10 in BM-MSC were associated with the inhibition of PHA-PBMC proliferation (sHLA-G, P = 0.0008, r = 0.9308; membrane HLA-G, P = 0.0005, r = 0.9502).
  
  
  CONCLUSIONS
  We propose the evaluation of sHLA-G production in IL-10-treated BM-MSC cultures as a possible marker of immunoregulatory function.
authors.affiliation: Department of Experimental and Diagnostic Medicine, Laboratory of Immunogenetics, Section of Medical Genetics, University of Ferrara, Ferrara, Italy. rbr@unife.it
authors.forename: Roberta
authors.initials: R
authors.lastname: Rizzo
authors.forename: Giacomo
authors.initials: G
authors.lastname: Lanzoni
authors.forename: Marina
authors.initials: M
authors.lastname: Stignani
authors.forename: Diana
authors.initials: D
authors.lastname: Campioni
authors.forename: Francesco
authors.initials: F
authors.lastname: Alviano
authors.forename: Francesca
authors.initials: F
authors.lastname: Ricci
authors.forename: Pier Luigi
authors.initials: PL
authors.lastname: Tazzari
authors.forename: Loredana
authors.initials: L
authors.lastname: Melchiorri
authors.forename: Sergio Zaccaria
authors.initials: SZ
authors.lastname: Scalinci
authors.forename: Antonio
authors.initials: A
authors.lastname: Cuneo
authors.forename: Laura
authors.initials: L
authors.lastname: Bonsi
authors.forename: Francesco
authors.initials: F
authors.lastname: Lanza
authors.forename: Gian Paolo
authors.initials: GP
authors.lastname: Bagnara
authors.forename: Olavio R
authors.initials: OR
authors.lastname: Baricordi
mesh_terms: D000328:Adult; D001854:Bone Marrow Cells / Q000187:drug effects / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques; D006680:HLA Antigens / Q000032:analysis* / Q000096:biosynthesis; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000032:analysis* / Q000096:biosynthesis; D006801:Humans; D007108:Immune Tolerance; D007165:Immunosuppression Therapy; D016753:Interleukin-10 / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000276:immunology*; D008875:Middle Aged; D010835:Phytohemagglutinins / Q000494:pharmacology; D017154:Stromal Cells / Q000187:drug effects / Q000276:immunology
chemical_list: D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D010835:Phytohemagglutinins; D016753:Interleukin-10