Induction of HLA-DR by interferon-gamma requires a trypsin-like protease.
Induction of HLA-DR antigen expression by interferon-gamma (IFN-gamma) is inhibited by trypsin inhibitors and an anti-trypsin monoclonal antibody, but not by chymotrypsin inhibitors, suggesting a requirement for trypsin-like protease (TLP) activity in IFN-gamma-induced HLA-DR expression. Using p-nitroanilide and thioester substrates, TLP activity was demonstrated in cellular extracts of a hybrid epidermal cell line and judged to be essential for HLA-DR expression. TLP activity was inhibited by the trypsin inhibitors soybean trypsin inhibitor, ovomucoid trypsin inhibitor, and tosyl-lysyl-chloromethyl ketone and by an anti-trypsin monoclonal antibody, closely paralleling inhibition of HLA-DR expression by such agents. TLP activity was enhanced by exposure to trypsin-linked agarose, indicating that the protease normally exists in an inactive form, perhaps in an enzyme-inhibitor complex or as an activatable proenzyme. Finding glucocorticoids (GC) to also inhibit IFN-gamma-induced HLA-DR expression and to regulate serine protease, especially urokinase plasminogen activator (uPA), activity raised the possibility of GC regulation of TLP activity. However, TLP activity was found to be constitutively expressed, regulated by neither GC nor IFN-gamma, nor was uPA activity involved in HLA-DR regulation. Trypsin inhibitors and GC also inhibited induction of intracellular 2',5'-oligoadenylate (2-5A) synthetase by IFN-gamma. Thus, TLP activity is required for IFN-gamma induction of HLA-DR and 2-5A synthetase.
Smith
B
B
Dermatology Service, Department of Veterans Affairs Medical Center, Martinez, CA 94553.
Merlin
G
G
Berman
B
B
D015088:2',5'-Oligoadenylate Synthetase / Q000037:antagonists & inhibitors*; D005455:Fluorescent Antibody Technique; D006684:HLA-DR Antigens / Q000096:biosynthesis*; D006801:Humans; D006822:Hybrid Cells; D006863:Hydrogen-Ion Concentration; D007371:Interferon-gamma / Q000494:pharmacology*; D011994:Recombinant Proteins; D014357:Trypsin / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors / Q000378:metabolism
D006684:HLA-DR Antigens; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors; D007371:Interferon-gamma; D015088:2',5'-Oligoadenylate Synthetase; D014357:Trypsin; D014568:Urokinase-Type Plasminogen Activator