interleukin-8 and Flavobacteriaceae-Infections

Interleukin-8 (IL-8), a CXC-type chemokine, plays a key role in acute inflammation by recruiting neutrophils in mammals. In the present study, the open reading frame (ORF) of IL-8, encoding 99 amino acids was cloned in mandarin fish, and its function in inflammation was investigated. The IL-8 contains four conserved cysteine residues, with the first two forming the CXC signature motif. The genomic sequence of mandarin fish IL-8 has four exons and three introns, a typical gene organization of the CXC chemokine. Bioactive recombinant IL-8 (rIL-8) exhibited a chemotactic effect on head kidney leukocytes in vitro, and activates the transcription of the inflammatory genes, IL-8 and IL-1β. When mandarin fish was challenged intraperitoneally with the pathogenic bacterium Flavobacterium columnare G

Topics: Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; DNA, Complementary; Fish Diseases; Fish Proteins; Flavobacteriaceae Infections; Flavobacterium; Head Kidney; Immunomodulation; Interleukin-1beta; Interleukin-8; Leukocytes; Perciformes; RNA, Messenger; Sequence Alignment; Transcription, Genetic

The mucosal immune systems of fishes are still poorly understood, and defined models for studying natural host-pathogen interactions are lacking. The objective of this study was to evaluate different challenge models and pathogens to examine the magnitude of change in intestinal immune gene expression. Rainbow trout were exposed by immersion to Yersinia ruckeri or by intraperitoneal injection with Flavobacterium psychrophilum. At 3, 9, or 10 days post-challenge, pathogen load was quantified by plate count and intestinal tissue was removed and immune gene expression measured by real-time PCR. In general, the magnitude of infection was correlated with change in immune gene transcript abundance. We found that messages for the innate immune molecules, SAA, IL-8, INF-γ and TNF-α, as well as the message for IgM, were up-regulated in intestinal tissue in both challenge paradigms. A >250-fold increase was observed in SAA and 20-fold increase of IL-8 gene transcript abundance occurred on day 10 following challenge with F. psychrophilum. Within individual fish, there was a positive correlation between bacteria load in the spleen and the increase of immune gene message between 3 and 10 days post-infection. These findings demonstrate that measurable changes in immune gene expression occur in the intestine of rainbow trout following bath challenge with Y. ruckeri or injection challenge with F. psychrophilum.

Topics: Animals; Fish Diseases; Flavobacteriaceae Infections; Immunity, Mucosal; Immunoglobulin M; Interferon-gamma; Interleukin-8; Oncorhynchus mykiss; Real-Time Polymerase Chain Reaction; Serum Amyloid A Protein; Yersinia Infections; Yersinia ruckeri

To further enhance our understanding of immunological gene expression in rainbow trout, Oncorhynchus mykiss, after infection with naturally occurring pathogens, a series of probes and primers were developed for the quantification of immune factors. Separate groups of specific-pathogen-free rainbow trout were infected with either Flavobacterium psychrophilum, Aeromonas salmonicida or infectious haematopoietic necrosis virus (IHNV). Three different concentrations of each pathogen were used and samples from infected and mock-infected fish were taken at either 1 or 5 days after infection. Ten fish were sampled at each time point for individual sections of liver, spleen and head kidney. Organ specimens from five of the fish were used to re-isolate and quantify the pathogen at the time the samples were taken. Total RNA was extracted from the organs of the remaining five animals. Using real-time polymerase chain reaction with fluorescent-labelled probes, the RNA from these organs was examined for the level of expression of the following immunological factors; an interferon related protein (MX-1), interleukin-8 (IL-8), the cytotoxic T-cell marker CD-8 and complement factor C3 (C3). They were also measured for the level of beta-actin, which was used as a standardization control for cellular RNA expression. Infection with IHNV produced the greatest change in expression level for all the immunological related factors examined in this study. IHNV elicited the best dose response profile, which was typically seen at 5-days post-infection for MX-1, C3, IL-8 and CD-8. Infection with A. salmonicida and F. psychrophilum showed elevated, but variable expression levels for several of the genes tested.

Topics: Actins; Aeromonas salmonicida; Animals; CD8 Antigens; Complement C3; DNA Primers; Fish Diseases; Flavobacteriaceae Infections; Flavobacterium; Gene Expression Profiling; Gram-Negative Bacterial Infections; Infectious hematopoietic necrosis virus; Interleukin-8; Kidney; Liver; Oncorhynchus mykiss; Reverse Transcriptase Polymerase Chain Reaction; Rhabdoviridae Infections; Sesquiterpenes; Specific Pathogen-Free Organisms; Spleen