title: Rapamycin-conditioned, alloantigen-pulsed dendritic cells promote indefinite survival of vascularized skin allografts in association with T regulatory cell expansion.
abstract: Clinically-applicable protocols that promote tolerance to vascularized skin grafts may contribute to more widespread use of composite tissue transplantation. We compared the properties of alloantigen (Ag)-pulsed, rapamycin (Rapa)-conditioned and control bone marrow-derived host myeloid dendritic cells (DCs) and their potential, together with transient immunosuppression (anti-lymphocyte serum+cyclosporine), to promote long-term, vascularized skin graft survival in Lewis rats across a full MHC barrier. Both types of DCs expressed low levels of CD86, but Rapa DC expressed lower levels of MHC II and CD40 and were less stimulatory in MLR. While both Rapa and control DCs produced low levels of IL-12p70 and moderate levels of IL-6 and IL-10 following TLR ligation, Rapa DC secreted significantly lower levels of IL-6 and IL-10 in response to LPS. Donor Ag-pulsed Rapa DC, but not control DC, induced long-term skin graft survival (median survival time >133 days) when administered 7 and 14 days post-transplant. Circulating T cells in hosts with long-surviving grafts were hyporesponsive to donor alloAg stimulation, but proliferated in response to third-party stimulation and produced IFN-gamma and IL-10. When recipients of long-surviving grafts were challenged with skin grafts, donor but not third-party grafts were prolonged, suggesting underlying regulatory mechanisms. Both flow cytometry and immunohistochemical analysis revealed that donor Ag-pulsed Rapa DC infusion expanded CD4+ Foxp3+ Treg in recipients' spleens, graft-associated lymph nodes and the graft. These data demonstrate for the first time that pharmacologically-modified, donor Ag-pulsed host DC administered post-transplant can promote indefinite vascularized skin graft survival, associated with Treg expansion.
authors.affiliation: Department of Surgery and Division of Plastic and Reconstructive Surgery, Pittsburgh, PA 15261, USA.
authors.forename: Elaine K
authors.initials: EK
authors.lastname: Horibe
authors.forename: Justin
authors.initials: J
authors.lastname: Sacks
authors.forename: Jignesh
authors.initials: J
authors.lastname: Unadkat
authors.forename: Giorgio
authors.initials: G
authors.lastname: Raimondi
authors.forename: Zhiliang
authors.initials: Z
authors.lastname: Wang
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Ikeguchi
authors.forename: Douglas
authors.initials: D
authors.lastname: Marsteller
authors.forename: Lydia M
authors.initials: LM
authors.lastname: Ferreira
authors.forename: Angus W
authors.initials: AW
authors.lastname: Thomson
authors.forename: W P Andrew
authors.initials: WP
authors.lastname: Lee
authors.forename: Maryam
authors.initials: M
authors.lastname: Feili-Hariri
mesh_terms: D000818:Animals; D019013:CD40 Antigens / Q000378:metabolism; D049109:Cell Proliferation; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D006085:Graft Survival / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007519:Isoantigens / Q000276:immunology*; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D011920:Rats, Inbred WF; D017207:Rats, Sprague-Dawley; D020123:Sirolimus / Q000494:pharmacology*; D012867:Skin / Q000098:blood supply / Q000276:immunology; D016038:Skin Transplantation / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology*; D019172:Transplantation Conditioning
chemical_list: D019013:CD40 Antigens; D000949:Histocompatibility Antigens Class II; D007166:Immunosuppressive Agents; D007519:Isoantigens; D020123:Sirolimus