A novel antagonist of CRTH2 blocks eosinophil release from bone marrow, chemotaxis and respiratory burst.
BACKGROUND
Chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) has been revealed to be a novel receptor for prostaglandin (PG) D(2), which is a major mast cell product released during the allergic response. The aim of this study was to analyze the effects of a newly developed small molecule antagonist of CRTH2, Cay10471, on eosinophil function with respect to recruitment, respiratory burst and degranulation.


METHODS
Chemotaxis of guinea pig bone marrow eosinophils and human peripheral blood eosinophils were determined using microBoyden chambers. Eosinophil release from bone marrow was investigated in the in situ perfused guinea pig hind limb preparation. Respiratory burst and degranulation were measured by flow cytometry.


RESULTS
Cay10471 bound with high affinity to recombinant human and guinea pig CRTH2, but not DP, receptors. The antagonist prevented the PGD(2)-induced release of eosinophils from guinea pig bone marrow, and inhibited the chemotaxis of guinea pig bone marrow eosinophils and human peripheral blood eosinophils. Pretreatment with PGD(2) primed eosinophils for chemotaxis towards eotaxin, and this effect was prevented by Cay10471. In contrast, PGD(2) inhibited the C5a-induced up-regulation of CD63, a cellular marker of degranulation, in a Cay10471-sensitive manner. Finally, Cay10471 abolished the respiratory burst of eosinophils upon stimulation by PGD(2).


CONCLUSIONS
These data further emphasize the importance of CRTH2 in eosinophil function and show that Cay10471 is a highly potent and selective antagonist of PGD(2)-induced eosinophil responses. Cay10471 might hence be a useful compound for the treatment of allergic diseases.
Royer
J F
JF
Institute of Experimental and Clinical Pharmacology, Medical University of Graz, Graz, Austria.
Schratl
P
P
Lorenz
S
S
Kostenis
E
E
Ulven
T
T
Schuligoi
R
R
Peskar
B A
BA
Heinemann
A
A
D000818:Animals; D001853:Bone Marrow; D002227:Carbazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D015550:Cell Degranulation; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D004804:Eosinophils / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D005260:Female; D006168:Guinea Pigs; D006801:Humans; D008297:Male; D011971:Receptors, Immunologic / Q000037:antagonists & inhibitors* / Q000502:physiology*; D011982:Receptors, Prostaglandin / Q000037:antagonists & inhibitors* / Q000502:physiology*; D016897:Respiratory Burst / Q000187:drug effects*; D013449:Sulfonamides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D018418:Th2 Cells / Q000378:metabolism
D002227:Carbazoles; D011971:Receptors, Immunologic; D011982:Receptors, Prostaglandin; D013449:Sulfonamides; C036293:prostaglandin D2 receptor