Protein kinase C mediates the signal for interferon-gamma mRNA expression in cytotoxic T cells after their adhesion to laminin.
A cytotoxic T-cell line, CTLL-2 cells, showed spreading after adhering to extracellular matrix proteins such as fibronectin (FN), laminin (LN) and hyarulonic acid (HA). The adhesion of CTLL-2 cells to LN was mediated by very late activation antigen-6 (VLA-6). Expression of interferon-gamma (IFN-gamma) mRNA was enhanced in CTLL-2 cells, also when they adhered to extracellular matrix proteins; and the enhanced IFN-gamma mRNA expression by adhering to LN was blocked by anti-alpha 6 antibody. Calphostin C, a protein kinase C (PKC) inhibitor, markedly inhibited the enhancement of IFN-gamma mRNA expression in a dose-dependent manner, which suggested that PKC acted as a second messenger in the IFN-gamma mRNA expression mediated by the interaction of VLA-6 with LN in CTLL-2 cells. Furthermore, confocal laser-microscopic analysis and Western blot analysis revealed that PKC-alpha was activated after CTLL-2 cells adhered to LN. PKC activity translocated from the cytosol fraction to the particulate fraction, after CTLL-2 cells adhered to LN. Altogether, we suggest that PKC plays an important role in the signal transduction for IFN-gamma mRNA expression after cytotoxic T cells adhere to LN.
Li
Y Q
YQ
Laboratory of Pathology, Hokkaido University School of Medicine, Sapporo, Japan.
Kobayashi
M
M
Yuan
L
L
Wang
J
J
Matsushita
K
K
Hamada
J I
JI
Kimura
K
K
Yagita
H
H
Okumura
K
K
Hosokawa
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD / Q000276:immunology; D015153:Blotting, Western; D002448:Cell Adhesion / Q000276:immunology; D002460:Cell Line; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D039503:Integrin alpha6; D039121:Integrin alpha6beta1; D016023:Integrins / Q000378:metabolism; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000235:genetics; D007797:Laminin / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D009281:Naphthalenes / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics; D017474:Receptors, Laminin / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D016326:Extracellular Matrix Proteins; D039503:Integrin alpha6; D039121:Integrin alpha6beta1; D016023:Integrins; D007797:Laminin; D009281:Naphthalenes; D012333:RNA, Messenger; D017474:Receptors, Laminin; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D011493:Protein Kinase C; C058819:calphostin C; D013755:Tetradecanoylphorbol Acetate