Transfusion of ethylene carbodiimide-fixed donor splenocytes prolongs survival of vascularized skin allografts

J Surg Res. 2018 Jan:221:343-352. doi: 10.1016/j.jss.2017.06.005. Epub 2017 Oct 19.

Abstract

Background: Allograft rejection is a major obstacle to the widespread clinical application of vascularized composite allotransplantation. Recent studies revealed a noncytoreductive strategy to protect allografts by the transfusion of ethylene carbodiimide-fixed donor splenocytes (ECDI-SPs). To determine whether this approach offers advantages in protecting skin allografts, we examined the immunological protection of infusing ECDI-SPs with a 30-d administration of rapamycin on the skin allografts of mice.

Materials and methods: C57BL/6 recipient mice received BALB/c donor full-thickness skin or vascularized skin transplants at day 0, along with the infusion of donor ECDI-SPs 7 d before and 1 d after allotransplantation and a 30-d course of rapamycin. Recipients received ECDI-untreated splenocytes or C3H allografts as controls. In vitro allostimulatory activity of ECDI-SPs and donor-specific ex vivo hyporesponsiveness were tested. Production of related cytokines (TGF-β, IL-10, IL-1β, and TNF-α) and expression of CD4+Foxp3+ regulatory T cells (Tregs) were also examined.

Results: Transfusion of ECDI-SPs combined with rapamycin significantly prolonged survival of full-thickness skin (median survival time [MST]: 28 d) and full-thickness skin allografts (MST: 71 d) compared with untreated splenocytes (MSTs: 11 d and 30 d) or C3H allografts (MSTs: 11 d and 38 d). This effect was accompanied by increased production of IL-10 and TGF-β, decreased production of IL-1β and TNF-α, and expansion of Tregs in vitro and in vivo.

Conclusions: ECDI-SP infusion combined with short-term rapamycin administration provides a promising approach to prolong the skin allograft survival.

Keywords: Mice; Skin transplantation; Tolerance; Vascularized composite allotransplantation.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Cell Transplantation / methods*
  • Cytokines / metabolism
  • Ethyldimethylaminopropyl Carbodiimide
  • Graft Rejection / immunology
  • Graft Rejection / prevention & control*
  • Immunosuppressive Agents / administration & dosage*
  • Male
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Sirolimus / administration & dosage*
  • Skin Transplantation*
  • Spleen / cytology
  • T-Lymphocytes, Regulatory
  • Transplantation, Homologous

Substances

  • Cytokines
  • Immunosuppressive Agents
  • Ethyldimethylaminopropyl Carbodiimide
  • Sirolimus