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The Journal of Immunology, 1998, 161: 6273-6279.
Copyright © 1998 by The American Association of Immunologists

Identification of a Truncated Form of the CC Chemokine CKß-8 Demonstrating Greatly Enhanced Biological Activity

Colin H. Macphee1,*, Edward R. Appelbaum{dagger}, Kyung Johanson{ddagger}, Kitty E. Moores*, Christina S. Imburgia§, Jim Fornwald{dagger}, Theo Berkhout*, Mary Brawner{dagger}, Pieter H. E. Groot*, Kevin O’Donnell{ddagger}, Daniel O’Shannessy{ddagger}, Gil Scott{ddagger} and John R. White§

* Department of Vascular Biology, SmithKline Beecham Pharmaceuticals, The Pinnacles, Harlow, Essex, United Kingdom; and Departments of {dagger} Gene Expression Sciences, {ddagger} Protein Biochemistry, and § Molecular and Cellular Immunology, SmithKline Beecham, King of Prussia, PA 19406

A new CC chemokine, designated CKß-8 or myeloid progenitor inhibitor factor-1, was recently identified in a large scale sequencing effort and was cloned from a human aortic endothelial library. CKß-8 cDNA encodes a signal sequence of 21 amino acids, followed by a 99-amino acid predicted mature form. CKß-8 was expressed and purified from a baculovirus insect cell expression system, which resulted in the identification of different N-terminal variants of the secreted chemokine. The three major forms (containing amino acids 1–99, 24–99, and 25–99 of the secreted chemokine) showed a large variation in potency. CKß-8 activated both monocytes and eosinophils to mobilize intracellular calcium; however, the shortest form of CKß-8 (25–99) was >2 orders of magnitude more potent than the longest form. Cross-desensitization experiments in both monocytes and eosinophils suggested that the CCR1 receptor was probably the predominant receptor that mediates this chemokine’s physiologic response. However, incomplete desensitization was encountered in both cell systems, suggesting involvement of an additional receptor(s). Interestingly, the short form of CKß-8 was the most potent chemotactic chemokine that we have ever evaluated in the monocyte system (EC50 = 54 pM). However, in contrast to its action on monocytes, CKß-8 was a very poor chemotactic factor for eosinophils.




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