title: Effects of immunosuppressants on induction of regulatory cells after intratracheal delivery of alloantigen.
abstract: BACKGROUND
  We previously reported that intratracheal delivery (ITD) of alloantigen generated regulatory cells in mice. Here, we examined the effect of various doses of conventional immunosuppressants (FK506, cyclosporine A, azathioprine, mycophenolate mofetil, and rapamycin) on inducing regulatory cells in our model.
  
  
  METHODS
  CBA mice (primary recipients) were given C57BL/6 splenocytes by ITD and either no additional treatment or various doses of an immunosuppressant. Seven days later, splenocytes from these mice were adoptively transferred into naive secondary CBA recipients that underwent C57BL/6 cardiac grafting the same day.
  
  
  RESULTS
  Adoptive transfer from primary recipients given ITD of splenocytes alone induced prolonged allograft survival in secondary recipients (median survival time [MST], 50 days), suggesting that regulatory cells were generated. When ITD of alloantigen was combined with daily administration of 0.1 mg/kg FK506 or 0.2 mg/kg rapamycin, graft survival was similarly prolonged (MST 55 and 50 days, respectively). When combined with 20 or 40 mg/kg MMF or 0.4 mg/kg rapamycin, the majority of recipients demonstrated indefinite survival (MST, >100 days in all groups). When ITD of alloantigen was combined with 0.3, 0.5, or 1.0 mg/kg FK506; 5, 10, or 25 mg/kg cyclosporine A; or 1.0 or 2.0 mg/kg azathioprine, allografts were rejected acutely (MST 7-13 days).
  
  
  CONCLUSIONS
  Generation of regulatory cells by ITD of alloantigen was facilitated by mycophenolate mofetil and high doses of rapamycin but abrogated by cyclosporine A, azathioprine, and high doses of FK506. Low doses of rapamycin and of FK506 did not interfere with generation of regulatory cells.
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Shibutani
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Inoue
authors.forename: Osamu
authors.initials: O
authors.lastname: Aramaki
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Akiyama
authors.forename: Kenji
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Motohide
authors.initials: M
authors.lastname: Shimazu
authors.forename: Masaki
authors.initials: M
authors.lastname: Kitajima
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Ikeda
authors.forename: Nozomu
authors.initials: N
authors.lastname: Shirasugi
authors.forename: Masanori
authors.initials: M
authors.lastname: Niimi
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D001379:Azathioprine / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D016572:Cyclosporine / Q000494:pharmacology; D003713:Dendritic Cells / Q000276:immunology; D006085:Graft Survival / Q000276:immunology; D007165:Immunosuppression Therapy; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007376:Interleukin-2 / Q000096:biosynthesis / Q000378:metabolism; D007519:Isoantigens / Q000008:administration & dosage* / Q000276:immunology*; D051379:Mice; D009173:Mycophenolic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D020123:Sirolimus / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000187:drug effects* / Q000276:immunology* / Q000378:metabolism; D016559:Tacrolimus / Q000494:pharmacology; D013997:Time Factors; D014019:Tissue Donors; D014132:Trachea / Q000276:immunology*; D014180:Transplantation
chemical_list: D007166:Immunosuppressive Agents; D007376:Interleukin-2; D007519:Isoantigens; D016572:Cyclosporine; D009173:Mycophenolic Acid; D001379:Azathioprine; D020123:Sirolimus; D016559:Tacrolimus