期刊
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
卷 29, 期 1, 页码 33-42出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.dci.2004.05.006
关键词
real-time PCR; immunostimulants; prophenoloxidase; beta-1,3-glucan binding protein; lobster; Homarus gammarus; granulocytes
This paper presents data from an investigation of the mode of action of five different crustacean immunostimulants presented to European lobster (Homarus gammarus) granulocytes cultured in vitro. The experiments were designed to test whether or not the immunostimulants could cause the short-term up-regulation of genes coding for immune proteins without causing the cells to degranulate. Quantitative measurements of mRNA transcript abundance were made using real-time PCR and it was first necessary to isolate the complete gene sequences coding for the proteins prophenoloxidase (proPO), beta-1,3-glucan binding protein (betaGBP) and beta-actin (beta-act) in the lobster. These sequences were used to design TaqMan(TM) primer and fluorescent probe sets. The presented data indicated that the majority of the tested immunostimulants did not induce the up-regulation of immune-related gene expression in the granulocytes in isolation. Alternative modes of action, including the in vivo up-regulation of gene expression in haemopoetic tissues, are discussed. (C) 2004 Elsevier Ltd. All rights reserved.
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