Characterization of a receptor for human monocyte-derived neutrophil chemotactic factor/interleukin-8.
Monocyte-derived neutrophil chemotactic factor/interleukin-8 (MDNCF/IL-8) is an 8,000-dalton protein produced by monocytes which exhibits activity as a chemoattractant for neutrophils with maximal activity achieved at a concentration of 50 ng/ml. This polypeptide has been iodinated by chloramine-T methodology (350 Ci/mM), and specific receptors for MDNCF/IL-8 have been detected on human neutrophils, U937 cells, THP-1 cells, and dimethyl sulfoxide-differentiated HL-60 cells. The binding of MDNCF/IL-8 to human neutrophils is not inhibited by interleukin-1 alpha, tumor necrosis factor-alpha, insulin, or epidermal growth factor. In addition, chemoattractants such as C5a, fMet-Leu-Phe, leukotriene B4, and platelet-activating factor fail to inhibit binding, suggesting that MDNCF/IL-8 utilizes a unique receptor. The receptor for MDNCF/IL-8 is apparently glycosylated since ligand binding is inhibited by the presence of wheat germ agglutinin, a lectin with a binding specificity for N-acetylglucosamine and neuraminic acid. Steady state binding experiments indicate Kd values of 4 and 0.5 nM and receptor numbers of 75,000 and 7,400 for human neutrophils and differentiated HL-60 cells, respectively. 125I-MDNCF/IL-8 bound to human neutrophils is rapidly internalized and subsequently released from cells as trichloroacetic acid-soluble radioactivity. Affinity labeling experiments suggest that the human neutrophil MDNCF/IL-8 receptor exhibits a mass of approximately 58,000 daltons.
Department of Biochemistry, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut 06877.
P M
PM
Grob
E
E
David
T C
TC
Warren
R P
RP
DeLeon
P R
PR
Farina
C A
CA
Homon
D000345:Affinity Labels; D000818:Animals; D001667:Binding, Competitive; D002454:Cell Differentiation; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002630:Chemotactic Factors / Q000378:metabolism* / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D016209:Interleukin-8; D007378:Interleukins / Q000378:metabolism* / Q000494:pharmacology; D007457:Iodine Radioisotopes; D007700:Kinetics; D037102:Lectins / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D009000:Monocytes / Q000378:metabolism*; D009504:Neutrophils / Q000502:physiology; D011971:Receptors, Immunologic / Q000378:metabolism*; D023062:Receptors, Interleukin-8A; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D013696:Temperature; D014909:Wheat Germ Agglutinins / Q000494:pharmacology
D000345:Affinity Labels; D002630:Chemotactic Factors; D006023:Glycoproteins; D016209:Interleukin-8; D007378:Interleukins; D007457:Iodine Radioisotopes; D037102:Lectins; D011971:Receptors, Immunologic; D023062:Receptors, Interleukin-8A; D011994:Recombinant Proteins; D014909:Wheat Germ Agglutinins