title: Characterization of Human CD8(+)TCR(-) Facilitating Cells In Vitro and In Vivo in a NOD/SCID/IL2rγ(null) Mouse Model.
abstract: CD8(+)/TCR(-) facilitating cells (FCs) in mouse bone marrow (BM) significantly enhance engraftment of hematopoietic stem/progenitor cells (HSPCs). Human FC phenotype and mechanism of action remain to be defined. We report, for the first time, the phenotypic characterization of human FCs and correlation of phenotype with function. Approximately half of human FCs are CD8(+)/TCR(-)/CD56 negative (CD56(neg)); the remainder are CD8(+)/TCR(-)/CD56 bright (CD56(bright)). The CD56(neg) FC subpopulation significantly promotes homing of HSPCs to BM in nonobese diabetic/severe combined immunodeficiency/IL-2 receptor γ-chain knockout mouse recipients and enhances hematopoietic colony formation in vitro. The CD56(neg) FC subpopulation promotes rapid reconstitution of donor HSPCs without graft-versus-host disease (GVHD); recipients of CD56(bright) FCs plus HSPCs exhibit low donor chimerism early after transplantation, but the level of chimerism significantly increases with time. Recipients of HSPCs plus CD56(neg) or CD56(bright) FCs showed durable donor chimerism at significantly higher levels in BM. The majority of both FC subpopulations express CXCR4. Coculture of CD56(bright) FCs with HSPCs upregulates cathelicidin and β-defensin 2, factors that prime responsiveness of HSPCs to stromal cell-derived factor 1. Both FC subpopulations significantly upregulated mRNA expression of the HSPC growth factors and Flt3 ligand. These results indicate that human FCs exert a direct effect on HSPCs to enhance engraftment. Human FCs offer a potential regulatory cell-based therapy for enhancement of engraftment and prevention of GVHD.
authors.affiliation: Institute for Cellular Therapeutics, University of Louisville, Louisville, KY.
authors.forename: Y
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Institute for Cellular Therapeutics, University of Louisville, Louisville, KY.
authors.forename: M J
authors.initials: MJ
authors.lastname: Elliott
authors.affiliation: Institute for Cellular Therapeutics, University of Louisville, Louisville, KY.
authors.forename: E S
authors.initials: ES
authors.lastname: Yolcu
authors.affiliation: Institute for Cellular Therapeutics, University of Louisville, Louisville, KY.
authors.forename: T O
authors.initials: TO
authors.lastname: Miller
authors.affiliation: Stem Cell Biology Program at the James Graham Brown Cancer Center, University of Louisville, Louisville, KY.
authors.forename: J
authors.initials: J
authors.lastname: Ratajczak
authors.affiliation: Regenerex, LLC, Louisville, KY.
authors.forename: L D
authors.initials: LD
authors.lastname: Bozulic
authors.affiliation: Institute for Cellular Therapeutics, University of Louisville, Louisville, KY.
authors.forename: Y
authors.initials: Y
authors.lastname: Wen
authors.affiliation: Institute for Cellular Therapeutics, University of Louisville, Louisville, KY.
authors.forename: H
authors.initials: H
authors.lastname: Xu
authors.affiliation: Stem Cell Biology Program at the James Graham Brown Cancer Center, University of Louisville, Louisville, KY.
authors.forename: M Z
authors.initials: MZ
authors.lastname: Ratajczak
authors.affiliation: Institute for Cellular Therapeutics, University of Louisville, Louisville, KY.|Regenerex, LLC, Louisville, KY.
authors.forename: S T
authors.initials: ST
authors.lastname: Ildstad
mesh_terms: D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D016827:CD8 Antigens / Q000378:metabolism*; D002478:Cells, Cultured; D006086:Graft vs Host Disease / Q000276:immunology* / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D053631:Interleukin Receptor Common gamma Subunit / Q000502:physiology*; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D023421:Models, Animal; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014019:Tissue Donors; D018183:Transplantation Chimera
chemical_list: D016827:CD8 Antigens; D053631:Interleukin Receptor Common gamma Subunit; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell
keywords: basic (laboratory) research/science; bone marrow/hematopoietic stem cell transplantation; cellular biology; immune regulation; immunosuppression/immune modulation; stem cells; tolerance: chimerism