Reduced GRK2 level in T cells potentiates chemotaxis and signaling in response to CCL4.
Chemokine receptors belong to the family of G-protein-coupled receptors (GPCR). Phosphorylation of GPCR by GPCR kinases (GRKs) is considered to play an important role in desensitization of these receptors. We have recently shown in patients with rheumatoid arthritis that the level of GRK2 in lymphocytes is reduced by approximately 50%. However, the physiological relevance of reduced GRK2 levels in lymphocytes is not known. Here, we investigated whether reduced GRK2 expression changes the chemotactic response of T cells to the chemokines CCL3, CCL4, and CCL5. Activated T cells from GRK2+/- mice, which have a 50% reduction in GRK2 protein levels, showed a significant 40% increase in chemotaxis toward the CCR5 ligand CCL4. In addition, chemotaxis toward the CCR1 and CCR5 ligands CCL3 and CCL5 was also increased. Binding of CCL4 to activated T cells from GRK2+/- and wild-type (WT) mice was similar, but agonist-induced CCR5 phosphorylation was attenuated in GRK2+/- cells. Moreover, the calcium response and phosphorylation of protein kinase B and extracellular-regulated kinase in response to CCL4 were significantly increased in GRK2+/- T cells, showing that signaling is increased when the level of GRK2 is reduced. GRK2+/- and WT cells do become refractory to restimulation with CCL4. In conclusion, a 50% decrease in T cell GRK2 expression results in increased responsiveness to CCL3, CCL4, and CCL5, suggesting that the 50% reduction in lymphocyte GRK2 level as observed during inflammation can have functional consequences for the response of these cells to chemokines.
Vroon
Anne
A
Laboratory for Psychoneuroimmunology, University Medical Center Utrecht, The Netherlands.
Heijnen
Cobi J
CJ
Lombardi
Maria Stella
MS
Cobelens
Pieter M
PM
Mayor
Federico
F
Caron
Marc G
MG
Kavelaars
Annemieke
A
D000818:Animals; D020013:Calcium Signaling; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D018946:Chemokine CCL5; D019742:Chemokines, CC / Q000378:metabolism / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte; D017868:Cyclic AMP-Dependent Protein Kinases / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D054769:G-Protein-Coupled Receptor Kinase 2; D006579:Heterozygote; D006801:Humans; D007700:Kinetics; D008213:Lymphocyte Activation; D019402:Macrophage Inflammatory Proteins; D051379:Mice; D018345:Mice, Knockout; D010766:Phosphorylation; D011506:Proteins / Q000494:pharmacology*; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000378:metabolism / Q000502:physiology*; D051552:beta-Adrenergic Receptor Kinases
C066507:CCL4 protein, human; C483024:CCL5 protein, human; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D018946:Chemokine CCL5; D019742:Chemokines, CC; D019402:Macrophage Inflammatory Proteins; D011506:Proteins; D017868:Cyclic AMP-Dependent Protein Kinases; C496567:GRK2 protein, human; D051552:beta-Adrenergic Receptor Kinases; D054769:G-Protein-Coupled Receptor Kinase 2