Apoptotic donor leukocytes limit mixed-chimerism induced by CD40-CD154 blockade in allogeneic bone marrow transplantation.
"Mini" allogeneic bone marrow transplants using non-myeloablative conditioning have reduced early treatment-related mortalities, but graft-versus-host disease (GVHD) and graft rejection remain clinical problems. Our preliminary studies indicated that low-dose busulfan conditioning and costimulatory blockade using anti-CD154 monoclonal antibody (mAb) in combination with a pretransplantation "tolerating" dose of bone marrow (BM) cells were sufficient to establish stable mixed-chimerism without GVHD when transplanting moderate doses of T cell depleted (TCD)-BM from major histocompatibility complex (MHC) fully-mismatched donors (Adams AB, Durham MM, Kean L, et al. J Immunol. 2001;167:1103-1111). In this study, donor splenocytes were administered before transplantation as a tolerating cell infusion with a conditioning regimen consisting of low-dose busulfan and anti-CD154 mAb. We compared the ability of viable and apoptotic donor cells of different ex vivo treatments and purified different donor cell populations (CD3(+), CD3(-), CD11b(+), and CD11b(-) splenocytes) to induce tolerance and enhance donor chimerism in a MHC mismatched model of murine bone marrow transplantation. We found that mixed chimerism without GVHD was enhanced by pretransplantation administration of viable allogeneic splenocytes and diminished in mice with prior exposure to apoptotic/necrotic donor splenocytes. CD11b(+)-enriched splenocytes more potently enhanced donor chimerism compared to unfractionated splenocytes or other splenocyte subsets. Mixed lymphocyte cultures demonstrated that apoptotic stimulators overcame the immune-tolerating activity of anti-CD154 mAb and led to increased interferon gamma and tumor necrosis factor alpha synthesis, increased proliferation of responder T cells, and decreased production of interleukin-10. In conclusion, viable donor splenocytes administered before transplantation in combination with costimulatory blockade induced tolerance and enhanced donor chimerism, whereas pretransplantation administration of apoptotic/necrotic donor cells led to host T cell activation and decreased overall donor engraftment.
Li
Jian-ming
JM
Division of Hematology and Oncology, Winship Cancer Institute, Emory University, Atlanta, Georgia 30322, USA.
Gorechlad
John
J
Larsen
Christian P
CP
Waller
Edmund K
EK
D000818:Animals; D020296:Animals, Congenic; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000494:pharmacology*; D017209:Apoptosis; D016026:Bone Marrow Transplantation / Q000473:pathology*; D002066:Busulfan / Q000008:administration & dosage / Q000494:pharmacology*; D019013:CD40 Antigens / Q000276:immunology*; D023201:CD40 Ligand / Q000276:immunology*; D017077:Culture Media, Conditioned / Q000737:chemistry; D006082:Graft Enhancement, Immunologic / Q000379:methods*; D006085:Graft Survival; D006086:Graft vs Host Disease / Q000517:prevention & control; D006183:H-2 Antigens / Q000276:immunology; D007371:Interferon-gamma / Q000032:analysis; D016753:Interleukin-10 / Q000032:analysis; D017708:Leukocyte Transfusion; D007962:Leukocytes / Q000166:cytology*; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000166:cytology; D018183:Transplantation Chimera; D019172:Transplantation Conditioning / Q000379:methods*; D023001:Transplantation Tolerance; D014184:Transplantation, Homologous / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
D000911:Antibodies, Monoclonal; D019013:CD40 Antigens; D017077:Culture Media, Conditioned; D006183:H-2 Antigens; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D023201:CD40 Ligand; D007371:Interferon-gamma; D002066:Busulfan