Regulation of galectin-9 expression and release in Jurkat T cell line cells.
Ecalectin/galectin-9 was recently described as a novel eosinophil chemoattractant highly expressed in immune tissues. We investigated the regulation of galectin-9 expression and release in Jurkat (a T cell line) cells. We demonstrated that medium and long-sized galectin-9 isoforms were constitutively expressed, and phorbol 12-myriastate 13-acetate (PMA) upregulated the level of galectin-9 mRNA in Jurkat cells. Western blotting and flow cytometry analyses revealed that PMA stimulation resulted in the upregulation of both intracellular and surface galectin-9 protein. The stimulated Jurkat cells simultaneously released evident eosinophil chemoattractant activity (ECA). Main ECA was adsorbed by both lactose and anti-galectin-9 antibody affinity column, suggesting that the ECA was ascribed to galectin-9. When Jurkat cells were stimulated with PMA in the presence of a BB94, a matrix metalloproteinase (MMP) inhibitor, but not tissue inhibitor of metalloproteinase-1 (TIMP-1), the release of galectin-9 was suppressed in a dose-dependent manner. We further found that calphostin c, a protein kinase c (PKC) inhibitor, weakly but significantly suppressed the release of galectin-9. The present data suggested that galectin-9 production in Jurkat cells is provoked by the stimulation with PMA and that some MMP and PKC is, at least, partly involved in the release of galectin-9 from Jurkat cells.
Chabot
Sophie
S
Department of Immunology and Immunopathology, Kagawa Medical University, 1750-1 Ikenobe, Miki-Cho, Kita-gun, Kagawa 761-0793, Japan.
Kashio
Yumiko
Y
Seki
Masako
M
Shirato
Yukako
Y
Nakamura
Kazuhiro
K
Nishi
Nozomu
N
Nakamura
Takanori
T
Matsumoto
Ryoji
R
Hirashima
Mitsuomi
M
D000818:Animals; D000917:Antibody Formation; D000970:Antineoplastic Agents / Q000494:pharmacology; D015153:Blotting, Western; D002273:Carcinogens / Q000494:pharmacology; D002631:Chemotactic Factors, Eosinophil / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D037161:Galectins; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D019169:Jurkat Cells / Q000378:metabolism*; D037102:Lectins / Q000235:genetics* / Q000378:metabolism; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D009281:Naphthalenes / Q000494:pharmacology; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects
D000970:Antineoplastic Agents; D002273:Carcinogens; D002631:Chemotactic Factors, Eosinophil; D017931:DNA Primers; D004791:Enzyme Inhibitors; D037161:Galectins; C105032:LGALS9 protein, human; D037102:Lectins; D009281:Naphthalenes; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013876:Thiophenes; D019715:Tissue Inhibitor of Metalloproteinase-1; D010649:Phenylalanine; C080985:batimastat; D011493:Protein Kinase C; D008666:Metalloendopeptidases; C058819:calphostin C; D013755:Tetradecanoylphorbol Acetate