Chemotaxins C5a and fMLP induce release of calprotectin (leucocyte L1 protein) from polymorphonuclear cells in vitro.
OBJECTIVE
To determine whether the chemotaxins C5a and formyl peptide (fMLP) can stimulate the release of calprotectin, the major leucocyte protein of polymorphonuclear neutrophils (PMN).


METHODS
A dose response curve for the uptake of 125I labelled rC5a and fMLP in PMN was determined by radioimmunoassay. The unlabelled chemotaxins were then incubated with PMN and the concentration of calprotectin in PMN lysates and supernatants was measured by an enzyme immunoassay.


RESULTS
Both rC5a and fMLP induced release of calprotectin from PMN in a dose dependent manner as determined by a reduction in intracellular calprotectin concentration. A minimum of approximately 10% of total PMN calprotectin was retained at concentrations of 10-100 nM of rC5a and 0.1-10.0 nM of fMLP. Antibodies to C5a reduced the rC5a mediated release of calprotectin, and the fMLP antagonist N-t-Boc-MLP inhibited the fMLP induced calprotectin release. Because receptors for rC5a (CD88) and fMLP are G protein coupled and thought to be pertussis toxin sensitive, PMN were incubated with this toxin before the experiments. The toxin was found to reduce uptake of rC5a by the cells and to inhibit rC5a and fMLP mediated calprotectin release.


CONCLUSIONS
rC5a and fMLP mediate release of calprotectin from PMN in vitro. This effect might be important during human infections in vivo.
Hetland
G
G
Institute of Immunology and Rheumatology, National Hospital, Oslo, Norway.
Talgö
G J
GJ
Fagerhol
M K
MK
D000954:Antigens, Surface / Q000097:blood; D002135:Calcium-Binding Proteins / Q000097:blood*; D018929:Cell Culture Techniques; D015936:Complement C5a / Q000276:immunology / Q000494:pharmacology*; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins / Q000097:blood*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D019006:Neural Cell Adhesion Molecules / Q000097:blood*; D018375:Neutrophil Activation / Q000502:physiology; D009504:Neutrophils / Q000378:metabolism*; D037342:Pertussis Toxin; D011994:Recombinant Proteins / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D000954:Antigens, Surface; D002135:Calcium-Binding Proteins; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins; D019006:Neural Cell Adhesion Molecules; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015936:Complement C5a; D037342:Pertussis Toxin