title: Pretreatment of transfused donor splenocytes and allografts with mitomycin C attenuates acute rejection in heart transplantation in mice.
abstract: OBJECTIVE
  The aim of this study was to investigate the effect of pretreatment of donor splenocytes and grafts with mitomycin C (MMC) on heart allograft survival, as well as to demonstrate the mechanism of function.
  
  
  METHODS
  Donor splenocytes from Balb/C mice were incubated with MMC (40 μg/mL) in vitro and then transfused into recipients (C57BL/6 mice). The heart allograft was perfused with MMC before harvest. Graft survival and histopathology were examined. Lymphocyte activation, regulatory T cells, and donor splenocyte apoptosis were examined with the use of flow cytometry.
  
  
  RESULTS
  MMC incubation in vitro induced apoptosis of donor splenocytes (15.5 ± 2.3% vs 23.2 ± 4.2%; P < .01). Either intravenous injection of MMC-treated donor splenocytes or transplantation of allograft pretreated with MMC prolonged heart allograft mean survival time from 7 ± 0.8 days to 20.5 ± 1.9 days or 10 ± 0.9 days, respectively (both P < .01). A combination of MMC-pretreated donor splenocyte transfusion and allografts showed the best effect on prolongation of graft survival (28.5 ± 1.8 days). Activation of CD4(+) T cells in spleen and peripheral lymph nodes of recipients was significantly inhibited by either MMC-splenocyte transfusion or the combination treatment. Meanwhile, the percentage of CD4(+)CD25(+)Foxp3(+) regulatory T cells in the spleen was increased in the MMC-splenocyte transfusion group (15.5 ± 1.1% vs 18.2 ± 0.9%; P < .05).
  
  
  CONCLUSIONS
  Both injection of MMC-conditioned donor splenocytes and MMC-conditioned allograft have effects on prolongation of heart allograft survival in mice, and MMC-conditioned donor splenocytes might play an essential role. MMC pretreatment induced regulatory T cells likely through induction of donor splenocyte apoptosis, and thus it inhibited T-cell activation.
authors.affiliation: Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: L
authors.initials: L
authors.lastname: Liu
authors.affiliation: Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.affiliation: Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: X
authors.initials: X
authors.lastname: Yuan
authors.affiliation: Department of Urology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: D
authors.initials: D
authors.lastname: Wang
authors.affiliation: Laboratory of Surgery, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: W
authors.initials: W
authors.lastname: Zeng
authors.affiliation: Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: X
authors.initials: X
authors.lastname: He
authors.affiliation: Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: C
authors.initials: C
authors.lastname: Wang
authors.affiliation: Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China. Electronic address: dengsx@mail.sysu.edu.cn.
authors.forename: S
authors.initials: S
authors.lastname: Deng
mesh_terms: D000208:Acute Disease; D064591:Allografts; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D006084:Graft Rejection / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D006085:Graft Survival / Q000187:drug effects; D006321:Heart / Q000187:drug effects*; D016027:Heart Transplantation / Q000009:adverse effects / Q000379:methods*; D008198:Lymph Nodes / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D008213:Lymphocyte Activation / Q000187:drug effects; D008297:Male; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016685:Mitomycin / Q000494:pharmacology*; D013154:Spleen / Q000187:drug effects* / Q000276:immunology / Q000473:pathology / Q000637:transplantation*; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D013997:Time Factors
chemical_list: D016685:Mitomycin