The G protein beta3 subunit splice variant Gbeta3-s causes enhanced chemotaxis of human neutrophils in response to interleukin-8

Naunyn Schmiedebergs Arch Pharmacol. 1999 Jul;360(1):27-32. doi: 10.1007/s002109900040.

Abstract

A C825T polymorphism was recently described in GNB3, the gene encoding the Gbeta3 subunit of heterotrimeric G proteins. The 825T allele is associated with the expression of a shorter splice variant (Gbeta3-s) and enhanced signal transduction via pertussis toxin (PTX)-sensitive G proteins. Given the pivotal role of G protein betagamma dimers in chemotaxis, we related the genotype at the GNB3 locus as a marker for Gbeta3-s expression to chemotaxis of human neutrophils in response to stimulation with interleukin-8 (IL-8). IL-8, which activates a CXC receptor coupled to PTX-sensitive G proteins, induced at 10 nM an enhanced maximum chemotaxis of neutrophils from individuals with TC/TT genotype compared to CC genotype. Furthermore, migration of neutrophils from 825T allele carriers was 2.5-fold higher at 0.1 nM and 1 nM IL-8. At these concentrations of IL-8, no significant chemotaxis was observed in neutrophils from homozygous C825 allele carriers, indicating a genotype-dependent, different potency of IL-8 to chemoattract neutrophils. In contrast, IL-8-induced Ca2+ signals and O2- generation were independent of genotype. The role of Gbeta3-s in enhanced chemotaxis could be confirmed by determination of chemotaxis of COS-7 cells following transfection with either Gbeta3-s or "wild-type" Gbeta3. Upon stimulation of the transfected cells with the chemoattractant lysophosphatidic acid (LPA), we observed an enhanced chemotactic response of Gbeta3-s-transfected compared to Gbeta3-transfected COS-7 cells, confirming that Gbeta3-s actually causes enhanced chemotaxis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Alternative Splicing
  • Animals
  • Anions
  • COS Cells
  • Calcium / metabolism
  • Chemotactic Factors / pharmacology*
  • Chemotaxis, Leukocyte / genetics*
  • GTP-Binding Proteins / genetics*
  • Humans
  • In Vitro Techniques
  • Interleukin-8 / pharmacology*
  • Lysophospholipids / pharmacology
  • Neutrophils / physiology*
  • Oxygen / metabolism
  • Transfection

Substances

  • Anions
  • Chemotactic Factors
  • Interleukin-8
  • Lysophospholipids
  • GTP-Binding Proteins
  • Oxygen
  • Calcium