Abstract
<p class="1Body">Neutrophils recognize and destroy pathogens through activation of the Toll like receptor (TLR) system as part of the inflammatory response of innate immunity. The expression and modulation of genes in the TLR signaling pathway in caprine blood neutrophils was investigated. Following initial screening for infection, goats (N=15) were assigned to three groups of five (n=5) individuals. Goats were drenched daily with 10 mL of powdered CorPet (Mycology labs Inc) soaked in hot (treatment I) or cold (treatment II) sterile filtered endotoxin free water, for a 4-week period. A control group of five age-matched goats received sterile water (treatment III). Blood was collected weekly and analyzed for packed cell volume and white blood cell differential counts. At weeks 1 and 4 neutrophils were isolated, using differential centrifugation and hypotonic lysis of red blood cells. The concentration and purity of total RNA isolated using Trizol was determined on a Nanodrop spectrophotometer. The RETROscript kit was used to synthesize cDNA. The expression of 84 genes in the human TLR signaling pathway RT<sup>2</sup> PCR Array was evaluated using real time PCR and the Livak method. The house keeping gene GAPDH was used to normalize the data. At week 1 untreated goats expressed 48 genes in the pathway. Goat neutrophils expressed 10 TLRs. Mushroom extracts modulated expression of and signaling by TLR. These results will help in the definition of the role of TLR expression in neutrophils and its contribution to goat innate immunity. Further this may aid in the design of therapeutics for goat health.</p>
Highlights
Goats live in a wide variety of microbe-rich environments
Primary sensing of pathogen associated molecular patterns (PAMPs) to alert the innate immune system is achieved by an array of germ-like encoded receptors known as pattern recognition receptors (PRRs) (Tirurugaan et al, 2010)
This study evaluated the expression and modulation of genes in the Toll like receptor (TLR) signaling pathway in goat neutrophils
Summary
Goats live in a wide variety of microbe-rich environments. It is crucial to have a sensitive innate defense mechanism which relies in part by recognizing conserved molecules that are unique to some classes of potential pathogens. The innate immune system is based principally on physical and chemical barriers to infection, as well as on different cell types recognizing invading pathogens and activating antimicrobial immune responses (Basset et al, 2003) They are key innate immune effector cells that provide early defense against invading microorganisms (Prince et al, 2011). Toll-like Receptors recognize microbial markers namely proteins, carbohydrates, lipids, nucleic acids and/or their combinations in an efficient, non-self-reactive manner to initiate a complex signaling cascade and activate a wide variety of transcription factors and inflammatory cytokines (Akira & Takeda, 2004). These cell surface molecules activate complement, phagocytosis, inflammation and apoptosis in response to pathogen jmbr.ccsenet.org
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