title: Isolation of regulatory T cells in the skin of a human hand-allograft, up to six years posttransplantation.
abstract: BACKGROUND
  A bilateral hand allotransplantation was performed in a patient six years ago. Whereas skin is known to be highly immunogenic, grafts have been well accepted up to now. Therefore, here we investigated the putative presence of regulatory T cells in the graft.
  
  
  METHODS
  Skin biopsies were performed at different time points and analyzed by immunochemistry. T cells were initially expanded with interleukin (IL)-2. In the latter biopsy, skin was directly analyzed by reverse-transcriptase polymerase chain reaction without any culture.
  
  
  RESULTS
  When tested against donor mononuclear cells, donor-primed skin T cells demonstrated unresponsiveness and inhibited donor-directed blood T cell alloresponse. Moreover, their T-cell receptor-Vbeta repertoire was skewed, in contrast to that of peripheral blood T cells. Retrospectively, nuclear FoxP3 expression in skin was measured by immunohistochemistry and was found positive at that time, but appeared to increase with time. This result was supported by the measurement of FoxP3 messenger RNA (mRNA) expression in the latter fresh biopsy, which showed higher levels than that of blood, together with no expression of perforin mRNA, but increased expression of transforming growth factor-beta and IL-10. No FoxP3 mRNA expression was found in the contralateral leg, due to the absence of T cell infiltrate.
  
  
  CONCLUSIONS
  This study shows the presence of the FoxP3 marker, in a well accepted human composite tissue allograft, up to six years posttransplantation. Because a suppressive cytokinic profile was also detected intragraft, in the absence of perforin mRNA expression, our data suggest that regulatory T cells could play a role in the long-term survival of this allograft.
authors.affiliation: Department of Immunogenomics, Mixed Unit HCL-BioMerieux, Hopital E. Herriot, Lyon, France. assia.eljaafari@chu-lyon.fr
authors.forename: Assia
authors.initials: A
authors.lastname: Eljaafari
authors.forename: Lionel
authors.initials: L
authors.lastname: Badet
authors.forename: Jean
authors.initials: J
authors.lastname: Kanitakis
authors.forename: Christophe
authors.initials: C
authors.lastname: Ferrand
authors.forename: Annie
authors.initials: A
authors.lastname: Farre
authors.forename: Palmina
authors.initials: P
authors.lastname: Petruzzo
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Morelon
authors.forename: Muriel
authors.initials: M
authors.lastname: Dubosson
authors.forename: Pierre
authors.initials: P
authors.lastname: Tiberghien
authors.forename: Valérie
authors.initials: V
authors.lastname: Dubois
authors.forename: Xavier
authors.initials: X
authors.lastname: Martin
authors.forename: Pierre
authors.initials: P
authors.lastname: Miossec
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Dubernard
mesh_terms: D000328:Adult; D051858:Forkhead Transcription Factors / Q000032:analysis / Q000235:genetics; D006085:Graft Survival / Q000276:immunology*; D063987:Hand Transplantation; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis; D012867:Skin / Q000737:chemistry / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D016212:Transforming Growth Factor beta / Q000235:genetics; D014184:Transplantation, Homologous
chemical_list: C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta; D016212:Transforming Growth Factor beta; D054353:Perforin; D016753:Interleukin-10