A fibronectin fragment induces tumor necrosis factor production of rat basophilic leukemia cells.
Proteolytic digest of fibronectin (FN), but not intact FN, induced TNF-alpha secretion of rat basophilic leukemia (RBL-2H3) cells. As a result of the identification of FN fragment responsible for TNF-alpha secretion, a 30-kDa fragment derived from the carboxyl-terminal heparin-binding (Hep 2) domain of FN was isolated from the FN digest. The TNF-alpha secretion was abrogated by treatment of RBL-2H3 cells with cycloheximide, indicating the de novo synthesis of TNF-alpha, but not with polymyxin B, excluding the possible TNF-alpha induction by some contaminated lipopolysaccharides. A 22-mer synthetic peptide originated from the Hep 2 domain, termed FNIII14, which has been found to negatively modulate the beta1 integrin activation, had the ability to induce TNF-alpha production, whereas this activity of FNIII14 disappeared by shuffling a YTIYVIAL sequence essential for the integrin-inactivating activity. FNIII14 suppressed the spreading of RBL-2H3 cells on FN substrate, wherein RBL-2H3 cell proliferation was inhibited with FNIII14 in a dose-dependent manner. Thus, it appears that FN fragments containing the YTIYVIAL anti-adhesive site affect the activation status of RBL-2H3 mast cells, characterized by the stimulation of TNF-alpha production and growth suppression, probably due to negative regulation of beta1 integrin activity.
Kamiya
Sadahiro
S
Department of Molecular Patho-Physiology, Faculty of Pharmaceutical Sciences, Tokyo University of Science (RIKADAI), 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan.
Kawaguchi
Tatsuya
T
Hasebe
Shinobu
S
Kamiya
Naoko
N
Saito
Yohei
Y
Miura
Shogo
S
Wada
Seiki
S
Yajima
Hirofumi
H
Katayama
Takashi
T
Fukai
Fumio
F
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D005353:Fibronectins / Q000494:pharmacology*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D015471:Leukemia, Basophilic, Acute / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008070:Lipopolysaccharides / Q000494:pharmacology; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology*; D011112:Polymyxin B / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D000900:Anti-Bacterial Agents; D005353:Fibronectins; D019012:Integrin beta1; D008070:Lipopolysaccharides; D010446:Peptide Fragments; D011500:Protein Synthesis Inhibitors; D014409:Tumor Necrosis Factor-alpha; D006493:Heparin; D003513:Cycloheximide; D011112:Polymyxin B