Cloning and pharmacological characterization of CCR7, CCL21 and CCL19 from Macaca fascicularis.
The chemokine receptor CCR7 and its ligands CCL19 and CCL21 play an important role in lymphocyte homing and have also been associated with inflammatory, allergic and lung disorders. Cloning of the cynomolgus monkey genes encoding CCR7, CCL19 and CCL21 revealed 93-97% sequence identity of the deduced proteins with their respective human homologs. In chemotaxis assays, B300-19 cells transfected with the cynomolgus (c) CCR7 receptor migrated in response to cCCL19 and cCCL21 in a dose-dependent manner with EC(50) values of 324+/-188nM and 247+/-29nM, respectively. cCCL19 and cCCL21 also elicited calcium responses in stable cell CHO-K1 lines expressing the cCCR7 receptor with EC(50) values of 227+/-4nM and 484+/-163nM, respectively. Although both human (h) CCL19 and hCCL21 elicited increases in intracellular calcium at the cCCR7 receptor, hCCL19 almost completely inhibited subsequent stimulation by hCCL21 whilst hCCL21 failed to inhibit subsequent stimulation by hCCL19. These results identify novel cynomolgus monkey genes and provide a model system for pre-clinical studies of potential drug candidates.
Moxley
Rachel
R
Novartis Horsham Reseach Centre, Wimblehurst Road, Horsham, West Sussex, RH12 5AB, UK.
Day
Elizabeth
E
Brown
Kate
K
Mahnke
Marion
M
Zurini
Mauro
M
Schmitz
Rita
R
Jones
Carol E
CE
Jarai
Gabor
G
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D054415:Chemokine CCL19 / Q000187:drug effects* / Q000235:genetics*; D054421:Chemokine CCL21 / Q000187:drug effects* / Q000235:genetics*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003001:Cloning, Molecular; D008252:Macaca fascicularis; D008969:Molecular Sequence Data; D054400:Receptors, CCR7 / Q000187:drug effects* / Q000235:genetics*; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D054415:Chemokine CCL19; D054421:Chemokine CCL21; D054400:Receptors, CCR7; D011994:Recombinant Proteins; D002118:Calcium