Real-time PCR quantification of the in vitro effects of crustacean immunostimulants on gene expression in lobster (Homarus gammarus) granular haemocytes

Christopher Hauton, JA Hammond, Valerie Jane Smith

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48 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)33-42
Number of pages10
JournalDevelopmental and Comparative Immunology
Volume29
Issue number1
DOIs
Publication statusPublished - Jan 2005

Keywords

  • real-time PCR
  • immunostimulants
  • prophenoloxidase
  • beta-1,3-glucan binding protein
  • lobster
  • Homarus gammarus
  • granulocytes
  • PROPHENOLOXIDASE-ACTIVATING SYSTEM
  • CRAYFISH PACIFASTACUS-LENIUSCULUS
  • BETA-1,3-GLUCAN BINDING-PROTEIN
  • FRESH-WATER CRAYFISH
  • MESSENGER-RNA LEVELS
  • BLACK TIGER SHRIMP
  • PENAEUS-MONODON
  • RT-PCR
  • LIPOPOLYSACCHARIDE
  • AQUACULTURE

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