title: Tissue inhibitor of metalloproteinase-3 regulates inflammation in human and mouse intestine.
abstract: OBJECTIVE
  Tissue inhibitor of metalloproteinases (TIMP)-3 is an inhibitor of matrix metalloproteinases, which regulates tissue inflammation, damage, and repair. We investigated the role of TIMP-3 in intestinal inflammation in human beings and mice.
  
  
  METHODS
  We used real-time polymerase chain reaction and flow cytometry to measure levels of TIMP-3 in intestine samples from patients with Crohn's disease (CD) and those without (controls). We also analyzed TIMP-3 levels in lamina propria mononuclear cells (LPMCs) collected from biopsy samples of individuals with or without CD (controls) and then stimulated with transforming growth factor (TGF)-β1, as well as in biopsy samples collected from patients with CD and then incubated with a Smad7 anti-sense oligonucleotide (knock down). LPMCs and biopsy samples from patients with CD were cultured with exogenous TIMP-3 and levels of inflammatory cytokines were measured. We evaluated the susceptibility of wild-type, TIMP-3-knockout (TIMP-3-KO), and transgenic (TIMP-3-Tg) mice to induction of colitis with 2, 4, 6-trinitrobenzene-sulfonic-acid (TNBS), and the course of colitis in recombinase-activating gene-1-null mice after transfer of wild-type or TIMP-3-KO T cells.
  
  
  RESULTS
  Levels of TIMP-3 were reduced in intestine samples from patients with CD compared with controls. Incubation of control LPMCs with TGF-β1 up-regulated TIMP-3; knockdown of Smad7, an inhibitor of TGF-β1, in biopsy samples from patients with CD increased levels of TIMP-3. Exogenous TIMP-3 reduced levels of inflammatory cytokines in CD LPMCs and biopsy samples. TIMP-3-KO mice developed severe colitis after administration of TNBS, whereas TIMP-3-Tg mice were resistant to TNBS-induced colitis. Reconstitution of recombinase-activating gene-1-null mice with T cells from TIMP-3-KO mice increased the severity of colitis, compared with reconstitution with wild-type T cells.
  
  
  CONCLUSIONS
  TIMP-3 is down-regulated in inflamed intestine of patients with CD. Its expression is regulated by TGF-β1, and knock-down of Smad7 in intestinal tissues from patient with CD up-regulates TIMP-3. Loss or reduction of TIMP-3 in mice promotes development of colitis.
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Ivan
authors: I
authors: Monteleone
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Massimo
authors: M
authors: Federici
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Massimiliano
authors: M
authors: Sarra
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Eleonora
authors: E
authors: Franzè
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Viviana
authors: V
authors: Casagrande
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Francesca
authors: F
authors: Zorzi
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Michele
authors: M
authors: Cavalera
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Angelamaria
authors: A
authors: Rizzo
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Renato
authors: R
authors: Lauro
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy.
authors: Francesco
authors: F
authors: Pallone
authors: Centre for Immunology and Infectious Disease, Blizard Institute of Cell and Molecular Science, Barts and the London School of Medicine and Dentistry, London, United Kingdom.
authors: Thomas T
authors: TT
authors: MacDonald
authors: Dipartimento di Medicina dei Sistemi, Università Tor Vergata, Rome, Italy. Electronic address: Gi.Monteleone@Med.uniroma2.it.
authors: Giovanni
authors: G
authors: Monteleone
mesh_terms: D000328:Adult; D000368:Aged; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D003092:Colitis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D003093:Colitis, Ulcerative / Q000378:metabolism*; D003424:Crohn Disease / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D015536:Down-Regulation; D055785:Gene Knockdown Techniques; D055786:Gene Knockout Techniques; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008875:Middle Aged; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism*; D051906:Smad7 Protein / Q000235:genetics; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014302:Trinitrobenzenesulfonic Acid
chemical_list: D016207:Cytokines; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D051906:Smad7 Protein; D019717:Tissue Inhibitor of Metalloproteinase-3; D016212:Transforming Growth Factor beta; D014302:Trinitrobenzenesulfonic Acid; D053829:Amyloid Precursor Protein Secretases