title: Canine bone marrow-derived mesenchymal stromal cells suppress alloreactive lymphocyte proliferation in vitro but fail to enhance engraftment in canine bone marrow transplantation.
abstract: Stable mixed hematopoietic chimerism has been consistently established in dogs who were mildly immunosuppressed by 200 cGy of total body irradiation (TBI) before undergoing dog leukocyte antigen (DLA)-identical bone marrow (BM) transplantation and who received a brief course of immunosuppression with mycophenolate mofetil (28 days) and cyclosporine (35 days) after transplantation. However, when TBI was reduced from 200 to 100 cGy, grafts were nearly uniformly rejected within 3-12 weeks. Here, we asked whether stable engraftment could be accomplished after a suboptimal dose of 100 cGy TBI with host immunosuppression enhanced by donor-derived mesenchymal stromal cells (MSCs) given after transplantation. MSCs were cultured from BM cells and evaluated in vitro for antigen expression. They showed profound immunosuppressive properties in mixed lymphocyte reactions (MLRs) in a cell dose-dependent manner not restricted by DLA. MSC and lymphocyte contact was not required, indicating that immunosuppression was mediated by soluble factors. Prostaglandin E2 was increased in culture supernatant when MSCs were cocultured in MLRs. The addition of indomethacin restored lymphocyte proliferation in cultures containing MSCs. MSCs expressed CD10, CD13, CD29, CD44, CD73/SH-3, CD90/Thy-1, and CD106/VCAM-1. For in vivo studies, MSCs were injected on the day of BM grafting and on day 35, the day of discontinuation of posttransplantation cyclosporine. MSCs derived from the respective BM donors failed to avert BM graft rejection in 4 dogs who received DLA-identical grafts after nonmyeloablative conditioning with 100 cGy TBI in a time course not significantly different from that of control dogs not given MSCs. Although the MSCs displayed in vitro characteristics similar to those reported for MSCs from other species, their immunosuppressive qualities failed to sustain stable BM engraftment in vivo in this canine model.
authors.affiliation: Transplantation Biology, Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Lee
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Scott S
authors.initials: SS
authors.lastname: Graves
authors.forename: Mineo
authors.initials: M
authors.lastname: Iwata
authors.forename: G M
authors.initials: GM
authors.lastname: Venkataraman
authors.forename: Marco
authors.initials: M
authors.lastname: Mielcarek
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Peterson
authors.forename: Susumu
authors.initials: S
authors.lastname: Ikehara
authors.forename: Beverly
authors.initials: B
authors.lastname: Torok-Storb
authors.forename: Rainer
authors.initials: R
authors.lastname: Storb
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D015703:Antigens, CD / Q000378:metabolism; D016026:Bone Marrow Transplantation; D002460:Cell Line; D049109:Cell Proliferation; D016572:Cyclosporine / Q000494:pharmacology; D004285:Dogs; D006084:Graft Rejection / Q000378:metabolism* / Q000473:pathology; D006085:Graft Survival; D015395:Histocompatibility Antigens Class I; D007165:Immunosuppression Therapy / Q000379:methods; D007166:Immunosuppressive Agents / Q000494:pharmacology; D008214:Lymphocytes / Q000378:metabolism* / Q000473:pathology; D009173:Mycophenolic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D017154:Stromal Cells / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D019172:Transplantation Conditioning; D014184:Transplantation, Homologous; D014916:Whole-Body Irradiation
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D015703:Antigens, CD; D015395:Histocompatibility Antigens Class I; D007166:Immunosuppressive Agents; C039692:histocompatibility antigen DLA; D016572:Cyclosporine; D009173:Mycophenolic Acid
references.citation: J Cell Biochem. 2006 Mar 1;97(4):744-54
references.pmid: 16229018
references.citation: Leukemia. 2007 Nov;21(11):2271-6
references.pmid: 17611560
references.citation: Stem Cells. 2001;19(1):46-58
references.pmid: 11209090
references.citation: Exp Mol Med. 2009 May 31;41(5):315-24
references.pmid: 19307751
references.citation: Haematologica. 2009 Feb;94(2):258-63
references.pmid: 19109217
references.citation: Exp Cell Res. 2004 May 1;295(2):395-406
references.pmid: 15093739
references.citation: Transplantation. 1996 Sep 27;62(6):876-7
references.pmid: 8824495
references.citation: Clin Immunol. 2008 Jun;127(3):348-58
references.pmid: 18387852
references.citation: Ann N Y Acad Sci. 2009 Sep;1176:124-34
references.pmid: 19796240
references.citation: Blood. 2007 Nov 15;110(10):3499-506
references.pmid: 17664353
references.citation: Cytotherapy. 2009;11(5):503-15
references.pmid: 19728189
references.citation: Blood. 1999 Oct 1;94(7):2523-9
references.pmid: 10498626
references.citation: Exp Hematol. 2009 Oct;37(10):1250-1257.e1
references.pmid: 19638293
references.citation: Nature. 2002 Jul 4;418(6893):41-9
references.pmid: 12077603
references.citation: Blood. 2004 Mar 1;103(5):1941-8
references.pmid: 14592834
references.citation: Biol Blood Marrow Transplant. 2005 May;11(5):389-98
references.pmid: 15846293
references.citation: Blood. 1998 Mar 15;91(6):1977-86
references.pmid: 9490680
references.citation: Transplantation. 2008 Apr 15;85(7):1023-9
references.pmid: 18408584
references.citation: Exp Hematol. 2000 Aug;28(8):875-84
references.pmid: 10989188
references.citation: Blood. 2002 May 15;99(10):3838-43
references.pmid: 11986244
references.citation: Stem Cells Dev. 2010 May;19(5):607-14
references.pmid: 19824807
references.citation: Exp Cell Res. 2005 Apr 15;305(1):33-41
references.pmid: 15777785
references.citation: Clin Orthop Relat Res. 2000 Oct;(379 Suppl):S71-90
references.pmid: 11039755
references.citation: Blood. 1998 May 1;91(9):3494-502
references.pmid: 9558410
references.citation: Proc Natl Acad Sci U S A. 1981 Dec;78(12):7327-30
references.pmid: 6174971
references.citation: Blood. 2007 Oct 1;110(7):2764-7
references.pmid: 17638847
references.citation: Lancet. 2008 May 10;371(9624):1579-86
references.pmid: 18468541
references.citation: Trends Immunol. 2002 Mar;23(3):144-50
references.pmid: 11864843
references.citation: Exp Hematol. 2002 Jan;30(1):42-8
references.pmid: 11823036
references.citation: Stem Cells. 2003;21(5):527-35
references.pmid: 12968107
references.citation: Science. 1999 Apr 2;284(5411):143-7
references.pmid: 10102814
references.citation: Biol Blood Marrow Transplant. 2003 Aug;9(8):489-95
references.pmid: 12931117
references.citation: Transplantation. 1994 Dec 27;58(12):1366-71
references.pmid: 7809930
references.citation: Transplantation. 2006 May 27;81(10):1390-7
references.pmid: 16732175
references.citation: Blood. 2005 Feb 15;105(4):1815-22
references.pmid: 15494428
references.citation: Haematologica. 2009 May;94(5):618-28
references.pmid: 19336739
references.citation: Blood. 1999 Aug 1;94(3):1131-6
references.pmid: 10419907
references.citation: Stem Cells. 2006 Nov;24(11):2582-91
references.pmid: 16873762
references.citation: J Clin Invest. 2000 Jun;105(12):1663-8
references.pmid: 10862779
references.citation: Cytotherapy. 2008;10(8):771-4
references.pmid: 19089685
references.citation: Stem Cells. 2006 Feb;24(2):386-98
references.pmid: 16123384
references.citation: Transplantation. 2003 Feb 15;75(3):389-97
references.pmid: 12589164
references.citation: Exp Hematol. 1983 Nov;11(10):961-6
references.pmid: 6420177
references.citation: Cytotherapy. 2008;10(7):676-85
references.pmid: 18985474
references.citation: Exp Hematol. 2008 Oct;36(10):1370-6
references.pmid: 18619727
references.citation: Blood. 1997 Apr 15;89(8):3048-54
references.pmid: 9108426
references.citation: Biol Blood Marrow Transplant. 2007 Feb;13(2):164-71
references.pmid: 17241922
references.citation: Bone Marrow Transplant. 2009 Feb;43(3):245-51
references.pmid: 18820709
references.citation: Tissue Antigens. 1998 Oct;52(4):397-401
references.pmid: 9820606
references.citation: Blood. 2007 Jul 1;110(1):418-23
references.pmid: 17369487
references.citation: Scand J Immunol. 2003 Jan;57(1):11-20
references.pmid: 12542793