transforming-growth-factor-beta and Protozoan-Infections--Animal

Expression of immune-regulatory genes that code for cyclooxigenase-2 (COX-2), transforming growth factor beta (TGF-beta), and two isoforms of interleukin-1beta (IL-1beta1 and IL-1beta2) was studied in susceptible and non-susceptible rainbow trout strains for 200 days after exposure to Myxobolus cerebralis. Expression of COX-2, IL-1beta1, and IL-1beta2 increased 5 min post exposure (p.e.) and was always more elevated in the susceptible strain than in the non-susceptible strain. In both strains, expression of COX-2 returned to the control level within a few hours p.e. Expression of IL-1beta1 and IL-1beta2 showed two elevated waves in both strains until 4 days p.e. Expression of TGF-beta in the non-susceptible strain was elevated at nearly all sampling points, but was decreased in the susceptible strain until up-regulation between 4 and 20 days p.e.; TGF-beta was the only gene where the expression in the non-susceptible strain was more elevated than in the susceptible strain. Rainbow trout of the non-susceptible strain appeared to resist infection by M. cerebralis with only minor transcriptional regulation of the genes investigated. Increased transcriptions of genes in the susceptible strain may be the result of an inability to antagonize the infection.

Topics: Animals; Cyclooxygenase 2; Disease Susceptibility; Eukaryota; Fish Diseases; Gene Expression Regulation; Interleukin-1beta; Oncorhynchus mykiss; Protein Isoforms; Protozoan Infections, Animal; Transforming Growth Factor beta

Amoebic gill disease (AGD) is an ectoparasitic disease caused by infection with the protozoan Neoparamoeba sp. and is characterised by epithelial hyperplasia that manifests as gill lesions. In order to examine the nature of the immune response to AGD, the expression of a range of immune-regulatory genes was examined in naïve uninfected rainbow trout, Oncorhynchus mykiss, and naïve rainbow trout subjected to a laboratory-induced AGD infection. The immune-regulatory genes examined were interleukin-1 beta isoform 1 (IL-1beta1), tumour necrosis factor alpha isoforms 1 and 2 (TNF-alpha1, TNF-alpha2), interleukin-8 (IL-8), transforming growth factor beta isoform 1 (TGF-beta1), inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), major histocompatibility complex beta chain (MHC-II beta-chain) and T-cell receptor beta chain (TCR beta-chain). Immune-regulatory genes that were up/down-regulated in AGD-infected trout compared to uninfected controls at 0, 7, and 14 days post-inoculation (p.i.) in gill, liver and anterior kidney tissue were initially identified by means of semi-quantitative RT-PCR. Up/down-regulated immune-regulatory genes were subsequently quantitated and validated by real-time RT-PCR (qRT-PCR). The extent of AGD-associated pathology was consistent amongst all AGD-infected trout at 7 days p.i. and increased considerably by 14 days p.i. At both 7 and 14 days p.i. IL-1beta1 and iNOS gene expression was significantly up-regulated in the gills, and IL-8 was significantly up-regulated in the liver of AGD-infected trout at 7 days p.i. These data demonstrate the involvement of the immune response to AGD at the molecular level, and indicate the importance of this response at the site of infection and the possible involvement of a systemic immune response.

Topics: Animals; DNA Primers; Fish Diseases; Gene Expression Regulation; Gills; Interleukin-1; Interleukin-8; Lobosea; Oncorhynchus mykiss; Protozoan Infections; Protozoan Infections, Animal; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha