Expression of the interleukin-8 receptors CXCR1 and CXCR2 on cord-blood-derived cultured human mast cells

Int Arch Allergy Immunol. 2002 Jun;128(2):142-50. doi: 10.1159/000059405.

Abstract

Background: An increase in mast cell number at sites of inflamed tissues has been observed. However, the expression of CXC chemokine receptors on human mast cells is poorly understood.

Methods: Cultured human mast cells were raised from human umbilical cord blood cells in the presence of stem cell factor and interleukin-6 (IL-6). The expression of surface chemokine receptors on the mast cells was analyzed by flow cytometry and that of mRNA was examined by the method of reverse transcriptase-polymerase chain reaction (RT-PCR). As functional assays for the receptors, mast cell migration was examined by a microchemotaxis assay and changes in the cytosolic free intracellular calcium concentrations ([Ca2+]i) was measured using fura-2-loaded mast cells, respectively.

Results: Expression of IL-8 receptors CXCR1 and CXCR2 was demonstrated by flow cytometry and of both mRNA by RT-PCR; however, CC chemokine receptors including CCR3 were not expressed on cord-blood-derived cultured human mast cells. IL-8 and its homologues showed chemotactic activity toward them in a dose-dependent manner, and IL-8 induced a dose-dependent rapid and transient increase in [Ca2+]i in the mast cells.

Conclusions: Our results suggest the surface expression of functional CXCR1 and CXCR2 on cord-blood-derived cultured human mast cells.

MeSH terms

  • Calcium / immunology
  • Chelating Agents / chemistry
  • Chemotaxis / immunology
  • Fetal Blood / cytology*
  • Flow Cytometry
  • Fura-2 / chemistry
  • Gene Expression Regulation / immunology*
  • Humans
  • Mast Cells / immunology*
  • Mast Cells / metabolism
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • Receptors, Interleukin-8A / biosynthesis*
  • Receptors, Interleukin-8A / genetics
  • Receptors, Interleukin-8A / immunology
  • Receptors, Interleukin-8B / biosynthesis*
  • Receptors, Interleukin-8B / genetics
  • Receptors, Interleukin-8B / immunology
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Chelating Agents
  • RNA, Messenger
  • Receptors, Interleukin-8A
  • Receptors, Interleukin-8B
  • Calcium
  • Fura-2