Inactivation of human anaphylatoxin C5a and C5a des-Arg through cleavage by the plasminogen activator activity of a human fibrosarcoma cell line.
The HT-1080 human fibrosarcoma cell line exhibited a plasminogen-dependent ability to inactivate recombinant anaphylatoxin C5a or zymosan-activated serum. The inactivation was obtained at physiological levels of both plasminogen (2 microM) and C5a (1-5 nM). Inactivated C5a and zymosan-activated serum were no longer able to induce chemotaxis and degranulation of neutrophils. Inactivation of C5a paralleled the emergence of plasmin activity, assayed by cleavage of the synthetic substrate H-D-valyl-L-leucyl-L-lysine-p-nitroanilide (S-2251). Both C5a inactivation and S-2251 cleavage were inhibited by the plasmin inhibitor alpha 2-antiplasmin, the urokinase inhibitor amiloride, and by anti-urokinase antibodies. In a cell-free system, inactivation of C5a was shown to depend on the simultaneous presence of urokinase and plasminogen and was inhibited by alpha 2-antiplasmin and by anti-urokinase antibodies. SDS-polyacrylamide electrophoresis demonstrated the cleavage of C5a by the plasminogen activation system and inhibition of the cleavage by amiloride. Amino acid sequencing of the band corresponding to the C5a degradation product revealed that C5a was cleaved at positions Lys14-His15 and Arg40-Ile41; cleavage at position Arg40-Ile41 seemed to be responsible for the loss of activity. Since neoplastic cells extensively produce and exhibit plasminogen activator activity, the present observations suggest that plasminogen activation may, by inactivation of C5a, reduce the anti-tumor immune response and support the immunological escape phenomenon of tumors.
Higazi A al-R
Department of Clinical Biochemistry, Hadassah University Hospital, Jerusalem, Israel.
Barghouti
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D000595:Amino Acid Sequence; D000706:Anaphylatoxins / Q000378:metabolism*; D002633:Chemotaxis / Q000502:physiology; D015936:Complement C5a / Q000378:metabolism*; D015937:Complement C5a, des-Arginine / Q000378:metabolism*; D005354:Fibrosarcoma / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010958:Plasminogen / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*; D015054:Zymosan / Q000494:pharmacology; D000979:alpha-2-Antiplasmin / Q000494:pharmacology
D000706:Anaphylatoxins; D015937:Complement C5a, des-Arginine; D000979:alpha-2-Antiplasmin; D015936:Complement C5a; D010958:Plasminogen; D015054:Zymosan; D014568:Urokinase-Type Plasminogen Activator