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J. Biol. Chem., Vol. 281, Issue 15, 9963-9970, April 14, 2006

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A Novel Hematopoietic Granulin Induces Proliferation of Goldfish (Carassius auratus L.) Macrophages^*

Patrick C. Hanington¹, Daniel R. Barreda¹, and Miodrag Belosevic^¶2

From the Departments of Biological Sciences and ^¶Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G E9, Canada and the Department of Hematology and Oncology, University of Pennsylvania, Philadelphia, Pennsylvania 19104

Granulins are a group of highly conserved growth factors that have been described from a variety of organisms spanning the metazoa. In this study, goldfish granulin was one of the most commonly identified transcripts in the differential cross-screening of macrophage cDNA libraries and was preferentially expressed in proliferating macrophages. Unlike mammalian granulins, which possess 7.5 repeats of a characteristic signature of 12 cysteine residues, the goldfish granulin encoded a putative peptide possessing only 1.5 cysteine repeats. Northern blot and real-time PCR analyses indicated that goldfish granulin was expressed only in the hematopoietic tissues of the goldfish, specifically the kidney and spleen, and in activated peripheral blood mononuclear cells. We expressed granulin using a prokaryotic expression system and produced an affinity-purified rabbit anti-goldfish granulin IgG. Recombinant goldfish granulin induced a dose-dependent proliferative response of goldfish macrophages that was inversely related to the myeloid differentiation stage of the cells studied. The highest proliferative response was observed in macrophage progenitor cells and monocytes. This proliferative response of macrophages was abrogated by the addition of anti-granulin IgG. These results indicate that goldfish granulin is a growth factor that positively modulates cell proliferation at distinct junctures of macrophage differentiation.

Received for publication, January 23, 2006 , and in revised form, February 9, 2006.

^* This work was supported in part by a grant from the Natural Sciences and Engineering Council of Canada (NSERC) (to M. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM//EBI Data Bank with accession number(s) DQ369750 [GenBank] .

¹ Supported by a NSERC doctoral scholarship.

² To whom correspondence should be addressed: Dept. of Biological Sciences, CW-405 Biological Sciences Bldg., University of Alberta, Edmonton, Alberta T6G 2E9, Canada. Tel.: 780-492-1266; Fax: 780-492-9234; E-mail: mike.belosevic{at}ualberta.ca.

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				P. C. Hanington, T. Wang, C. J. Secombes, and M. Belosevic Growth Factors of Lower Vertebrates: CHARACTERIZATION OF GOLDFISH (CARASSIUS AURATUS L.) MACROPHAGE COLONY-STIMULATING FACTOR-1 J. Biol. Chem., November 2, 2007; 282(44): 31865 - 31872. [Abstract] [Full Text] [PDF]