Cloning, expression, and functional characterization of cynomolgus monkey (Macaca fascicularis) CC chemokine receptor 1.
The CC chemokine receptor 1 (CCR1) has emerged as a relevant factor contributing to inflammatory diseases such as allergic asthma. Commonly used animal models of allergic airway inflammation, especially murine models, have certain limitations. The elaborate, nonhuman, primate models of asthma display the highest comparability with the situation in humans. These models play an important role in the understanding of the pathogenesis of asthma. To improve the understanding in cynomolgus monkey models, we identified and characterized CCR1 in this nonhuman primate. Initially, we cloned the cynomolgus monkey CCR1 (cCCR1) gene, and the sequence analysis revealed high homology at the nucleotide (92%) and amino acid (88.4%) levels with its human counterpart. Human embryonic kidney 293 cells were stably transfected with cCCR1 and used in functional assays. Among those CCR1 ligands tested, CCL14(9-74) was most potent in the induction of intracellular Ca2+ fluxes as observed for human CCR1 (hCCR1). Complete cross-desensitization could be achieved between CCL14(9-74) and CCL15. However, CCL3 could not fully abrogate the response to the potent ligand CCL14(9-74). Competition-binding studies with radiolabeled CCL3 concordantly showed that CCL14(9-74) has a higher affinity to cCCR1 than hCCL3. Moreover, differential tissue-specific expression of cCCR1 was investigated by real-time quantitative polymerase chain reaction, displaying the highest levels in spleen. This study adds basic information needed for the evaluation of the role of CCR1 in the pathophysiology of asthma using the highly relevant cynomolgus monkey model and in addition, aids in the preclinical evaluation of potential novel drugs targeting CCR1.
Gupta
Shipra
S
Department of Dermatology and Allergology, An-Institut, Hannover Medical School, Germany.
Schulz-Maronde
Sandra
S
Kutzleb
Christian
C
Richter
Rudolf
R
Forssmann
Wolf-Georg
WG
Kapp
Alexander
A
Forssmann
Ulf
U
Elsner
Jörn
J
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D018925:Chemokines / Q000494:pharmacology; D019742:Chemokines, CC / Q000494:pharmacology; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D008024:Ligands; D008252:Macaca fascicularis / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D054389:Receptors, CCR1; D019707:Receptors, Chemokine / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013997:Time Factors
C515300:CCR1 protein, human; C515301:Ccr1 protein, mouse; C515302:Ccr1 protein, rat; D018925:Chemokines; D019742:Chemokines, CC; D008024:Ligands; D054389:Receptors, CCR1; D019707:Receptors, Chemokine; D019206:GTP-Binding Protein alpha Subunits, Gi-Go