title: Increased levels of survivin, via association with heat shock protein 90, in mucosal T cells from patients with Crohn's disease.
abstract: OBJECTIVE
  Defective apoptosis of lamina propria T cells (LPTs) is involved in the pathogenesis of Crohn's disease. Survivin, a member of the inhibitors of apoptosis family, prevents cell death and regulates cell division. Survivin has been studied extensively in cancer, but little is known about its role in Crohn's disease.
  
  
  METHODS
  LPTs were isolated from mucosal samples of patients with Crohn's disease or ulcerative colitis and healthy individuals (controls). LPTs were activated with interleukin-2 or via CD3, CD2, and CD28 signaling, and cultured at 42°C to induce heat shock. Survivin expression was assessed by immunohistochemistry, confocal microscopy, and immunoblotting; survivin levels were reduced by RNA interference. Cell viability, apoptosis, and proliferation were measured by trypan blue exclusion, annexin-V/7-Aminoactinomycin D staining, and uptake of [3]thymidine, respectively.
  
  
  RESULTS
  LPTs from patients with Crohn's disease had higher levels of survivin than LPTs from patients with ulcerative colitis or controls. RNA knockdown of survivin in LPTs inhibited their proliferation and promoted apoptosis. Levels of survivin were low in LPTs from patients with ulcerative colitis and controls as a result of ubiquitin-mediated proteasome degradation. In LPTs from patients with Crohn's disease, survivin bound to the heat shock protein (HSP)90, and therefore was resistant to proteasome degradation. Incubating LPTs with 17-N-allylamino-17-demethoxygeldanamycin, an inhibitor of HSP90, reduced levels of survivin and induced apoptosis.
  
  
  CONCLUSIONS
  Levels of survivin are increased in LPTs from patients with Crohn's disease (compared with ulcerative colitis and controls) because survivin interacts with HSP90 and prevents proteasome degradation. This allows LPTs to avoid apoptosis. Strategies to restore apoptosis to these cells might be developed to treat patients with Crohn's disease.
authors: Department of Internal Medicine, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors: Heitor S P
authors: HS
authors: de Souza
authors: Gail A
authors: GA
authors: West
authors: Nancy
authors: N
authors: Rebert
authors: Carol
authors: C
authors: de la Motte
authors: Judy
authors: J
authors: Drazba
authors: Claudio
authors: C
authors: Fiocchi
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D051017:Apoptosis Regulatory Proteins; D018801:CD2 Antigens / Q000378:metabolism; D018106:CD28 Antigens / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003093:Colitis, Ulcerative / Q000276:immunology / Q000378:metabolism*; D003424:Crohn Disease / Q000276:immunology / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D019286:Endopeptidase K / Q000494:pharmacology; D005260:Female; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000378:metabolism*; D007376:Interleukin-2 / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000276:immunology / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D024101:Mitochondrial Proteins; D010766:Phosphorylation / Q000187:drug effects; D059748:Proteolysis; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D000077022:Survivin; D013601:T-Lymphocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D054875:Ubiquitination; D055815:Young Adult
chemical_list: C513674:Apopt1 protein, mouse; D051017:Apoptosis Regulatory Proteins; C107489:BIRC5 protein, human; D018801:CD2 Antigens; D018106:CD28 Antigens; D017252:CD3 Complex; D018841:HSP90 Heat-Shock Proteins; D051034:Inhibitor of Apoptosis Proteins; D007376:Interleukin-2; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D000077022:Survivin; D019286:Endopeptidase K