title: Effect of immunosuppressants on T-cell subsets observed in vivo using carboxy-fluorescein diacetate succinimidyl ester labeling.
abstract: BACKGROUND
  The in vivo effects of immunosuppressants on T cells are classically determined using animal models of organ transplantation. These methods are technically difficult and time consuming. A simple in vivo method is needed for screening new immunosuppressants.
  
  
  METHODS
  Donor mouse spleen cells were labeled with a fluorescent dye, carboxy-fluorescein diacetate succinimidyl ester (CFSE), and then injected into the blood of recipient severe combined immunodeficiency mice. Three days after the injection, spleen cells of the recipient mice were isolated and the proliferating alloreactive T cells were analyzed by flow cytometry.
  
  
  RESULTS
  In control recipient mice, 50% of the T cells were proliferating, consisting of both CD4+ and CD8+ T cells. In cyclosporine- or FK506-treated mice, T-cell proliferation was suppressed in the CD4 subset but not in the CD8 subset. On the contrary, T-cell proliferation was significantly reduced in the CD8 subset but not in the CD4 subset in recipient mice treated with rapamycin.
  
  
  CONCLUSIONS
  The present mouse model using carboxy-fluorescein diacetate succinimidyl ester labeling is simple and fast. It is useful for screening new immunosuppressants and for examining the effect on T-cell subsets.
authors.affiliation: Division of Transplantation, Department of Surgery, CSC Room H4/735, University of Wisconsin, 600 Highland Avenue, Madison, WI 53792. hu@surgery.wisc.edu
authors.forename: Huaizhong
authors.initials: H
authors.lastname: Hu
authors.forename: Yinchen
authors.initials: Y
authors.lastname: Dong
authors.forename: Ping
authors.initials: P
authors.lastname: Feng
authors.forename: John
authors.initials: J
authors.lastname: Fechner
authors.forename: Majed
authors.initials: M
authors.lastname: Hamawy
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Knechtle
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects*; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D016572:Cyclosporine / Q000494:pharmacology*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D020123:Sirolimus / Q000494:pharmacology*; D013388:Succinimides; D016559:Tacrolimus / Q000494:pharmacology*
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007166:Immunosuppressive Agents; D013388:Succinimides; D016572:Cyclosporine; D020123:Sirolimus; D016559:Tacrolimus