Identification and characterization of specific receptors for monocyte-derived neutrophil chemotactic factor (MDNCF) on human neutrophils.
Specific receptors for a recently purified and cloned monocyte-derived neutrophil chemotactic factor (MDNCF) have been identified on the surface of normal human peripheral blood neutrophils using 125I-labeled recombinant human MDNCF (125I-MDNCF). Competitive binding of 125I-MDNCF to human neutrophils reached a maximal level at 1-3 h at 4 degrees C. The Scatchard analysis showed that there are approximately 20,000 receptors per cell with a single type of high affinity binding (Kd, 8 x 10(-10) M). The receptors for MDNCF are clearly distinct from the receptors for other cytokines and chemotactic agents, e.g., IL-1 alpha, TNF-alpha, and FMLP, C5a, leukotriene B4, and platelet activating factor. Based on the SDS-PAGE analysis of chemically crosslinked 125I-MDNCF receptor complex, there are two polypeptides that bind MDNCF; the molecular weight of these two MDNCF receptors were estimated to be 67,000 and 59,000. Treatment of a promyelocytic cell line, HL60, with 1.25% DMSO for 5 d in vitro increased the number of receptors up to 7,000 receptors/cell with a Kd of 1.2 x 10(-9) M.
Samanta
A K
AK
Laboratory of Molecular Immunoregulation, National Cancer Institute, Frederick, Maryland 21701.
Oppenheim
J J
JJ
Matsushima
K
K
D002630:Chemotactic Factors / Q000378:metabolism*; D003432:Cross-Linking Reagents; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D016209:Interleukin-8; D007457:Iodine Radioisotopes; D007700:Kinetics; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism; D008970:Molecular Weight; D009000:Monocytes; D009504:Neutrophils / Q000032:analysis*; D011971:Receptors, Immunologic / Q000032:analysis* / Q000187:drug effects / Q000378:metabolism; D023062:Receptors, Interleukin-8A; D011994:Recombinant Proteins / Q000378:metabolism; D013388:Succinimides; D014407:Tumor Cells, Cultured
D002630:Chemotactic Factors; D003432:Cross-Linking Reagents; D016209:Interleukin-8; D007457:Iodine Radioisotopes; D011971:Receptors, Immunologic; D023062:Receptors, Interleukin-8A; D011994:Recombinant Proteins; D013388:Succinimides; C019358:disuccinimidyl suberate; D004121:Dimethyl Sulfoxide