Expression and functional activity of the IL-8 receptor type CXCR1 and CXCR2 on human mast cells

J Immunol. 1998 Sep 1;161(5):2600-8.

Abstract

To further elucidate mechanisms involved in mast cell accumulation at sites of cutaneous inflammation, we have studied the ability of human leukemic mast cells (HMC-1 cells) to express functionally active IL-8 receptors. Expression of mRNA for both types of IL-8 receptors (CXCR1 and CXCR2) was demonstrated by PCR and of both proteins by flow cytometry. Binding and competition studies with 125I-labeled IL-8 and its homologue melanoma growth stimulating activity (125I-labeled MGSA) revealed two specific binding sites for IL-8, K1 = 1.1 x 10(11) M(-1) and K2 = 5 x 10(7) M(-1); and for MGSA, K1 = 2.8 x 10(10) M(-1) and K2 = 5 x 10(7) M(-1). This finding was supported by a dose-dependent rise of cytosolic free calcium concentration ([Ca2+]i) induced by both chemokines and to a lesser extent by the homologue neutrophil-activating peptide-2 (NAP-2). A significant migratory response of human leukemic mast cells (HMC-1) was observed with all three chemokines at a range from 10(-8) M to 10(-9) M. Moreover, the formation of cellular F-actin was induced in a rapid, dose-dependent fashion, with a maximally 1.7-fold increase at 10(-7) M. Using postembedding immunoelectron microscopy, we could show the expression of CXCRI on the cytoplasmatic membrane of isolated human skin mast cells whereas CXCR2 was located in mast cell-specific granules. These findings demonstrate for the first time the functional expression of both types of IL-8 receptors on human mast cells, suggesting a role for their ligands during mast cell activation and recruitment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Antigens, CD / biosynthesis*
  • Antigens, CD / genetics
  • Antigens, CD / physiology
  • Antigens, CD / ultrastructure
  • Binding, Competitive
  • Calcium / metabolism
  • Cell Movement / drug effects
  • Chemokine CXCL1
  • Chemokines, CXC*
  • Chemotactic Factors / metabolism
  • Chemotactic Factors / pharmacology
  • Flow Cytometry
  • Growth Substances / metabolism
  • Growth Substances / pharmacology
  • HL-60 Cells
  • Humans
  • Intercellular Signaling Peptides and Proteins*
  • Interleukin-8 / metabolism*
  • Interleukin-8 / pharmacology
  • Iodine Radioisotopes
  • Mast Cells / metabolism*
  • Mast Cells / physiology
  • Mast Cells / ultrastructure
  • Peptides / pharmacology
  • Polymerase Chain Reaction
  • Protein Binding
  • RNA, Messenger / biosynthesis
  • Receptors, Chemokine / biosynthesis*
  • Receptors, Chemokine / genetics
  • Receptors, Chemokine / physiology
  • Receptors, Interleukin / biosynthesis*
  • Receptors, Interleukin / genetics
  • Receptors, Interleukin / physiology
  • Receptors, Interleukin / ultrastructure
  • Receptors, Interleukin-8A
  • Receptors, Interleukin-8B
  • Skin / metabolism
  • Skin / ultrastructure
  • Tumor Cells, Cultured
  • beta-Thromboglobulin

Substances

  • Actins
  • Antigens, CD
  • CXCL1 protein, human
  • Chemokine CXCL1
  • Chemokines, CXC
  • Chemotactic Factors
  • Growth Substances
  • Intercellular Signaling Peptides and Proteins
  • Interleukin-8
  • Iodine Radioisotopes
  • PPBP protein, human
  • Peptides
  • RNA, Messenger
  • Receptors, Chemokine
  • Receptors, Interleukin
  • Receptors, Interleukin-8A
  • Receptors, Interleukin-8B
  • beta-Thromboglobulin
  • connective tissue-activating peptide
  • Calcium