Production of recombinant C5a from rainbow trout (Oncorhynchus mykiss): role in leucocyte chemotaxis and respiratory burst.
Activation of the complement system can lead to the formation of the membrane attack complex, in which the component C5 is cleaved into C5a and C5b fragments. The C5a anaphylatoxin is a very potent pro-inflammatory molecule that induces chemotaxis and respiratory burst processes in a variety of mammalian leucocytes. While C5a has been well studied in mammals, little is known about the structure and function of C5a in teleost fish or other non-mammalian species. In the present study, we have produced and purified recombinant rainbow trout C5a (rtC5a), and we have shown that it plays an important role in inducing leucocyte migration as well as in triggering the respiratory burst of peripheral blood (PBLs) and head kidney leucocytes (HKLs). When the carboxy-terminal Arg was removed from rtC5a, its ability to induce cell migration and superoxide production remained intact. Interestingly, we show that leucocytes migrating towards rtC5a attached to the plate with a well-spread circular morphology, whereas those migrating towards activated trout serum displayed more irregular and dendritic-like shapes. Our data suggest that the basic mechanisms of action of the C5a anaphylotoxin have remained conserved for more than 300 million years.
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 413 Rosenthal, 3800 Spruce Street, Philadelphia, PA 19104, USA.
Hani
H
Boshra
Rodney
R
Peters
Jun
J
Li
J Oriol
JO
Sunyer
D000818:Animals; D002633:Chemotaxis / Q000187:drug effects*; D015936:Complement C5a / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000378:metabolism; D007962:Leukocytes / Q000187:drug effects*; D017686:Oncorhynchus mykiss / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000633:toxicity*; D016897:Respiratory Burst / Q000187:drug effects*
D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D015936:Complement C5a; D005982:Glutathione Transferase