Activation of carp leukocytes by a galactose-binding protein from Aphanomyces piscicida

Developmental and Comparative Immunology
Osamu Kurata, Kishio Hatai

Abstract

This study demonstrated that a galactose-binding protein (GBP) produced by a fish pathogenic water mold, Aphanomyces piscicida, activates carp leukocytes. Leukocytes were separated from the head kidney and peripheral blood using Percoll density centrifugation. A flow cytometric analysis revealed that GBP binds with many cells and a variety of cell types including lymphocytes, granulocytes and thrombocytes. Intracellular calcium flux of the peripheral blood leukocytes induced by stimulation with GBP was confirmed by counting the fluo-3 loaded cells whose fluorescence increased after the stimulation using flow cytometry. The percentage of cells in which a calcium flux was induced peaked 1 min after the stimulation. Approximately 6% of the cells specifically responded 1 min after the stimulation. The proliferation response was determined by the level of BrdU uptake by the leukocytes after the stimulation. Cell proliferation was observed 2, 4 and 6 days after stimulation with GBP. The expression of cytokines IL-1beta and TGF-beta1 in the peripheral blood leukocytes, after the stimulation was evaluated by a semi-quantitative reverse-transcriptase polymerase chain reaction. Increased expression of IL-1beta was observed 4h after stimu...Continue Reading

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Citations

Oct 16, 2007·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·Alexandre F S QueirozMaurício P de-Sales
Oct 6, 2006·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·Raniere M MouraMaurício P Sales
Jun 28, 2005·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·Suvendu DuttaShibnath Mazumder

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