Functional characterization of human and chicken TLR3
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Toll-like receptor 3 (TLR3) is a critical component of the innate immune system, responsible for detecting viral double-stranded RNA (dsRNA) and initiating downstream signaling pathways, including activation of the transcription factor NF-κB. While TLR3 function has been extensively characterized in humans and other mammals, little is known about its activity in avian species such as chickens, which are highly susceptible to RNA viruses like avian influenza. Chickens rely heavily on innate immune responses due to their limited adaptive immunity, making TLR3 an important mediator of early antiviral defense. Notably, naturally occurring polymorphisms in chicken TLR3 (chTLR3) have been identified, but their functional significance remains unexplored. This thesis sets the foundation for investigating the impact of chTLR3 polymorphisms on receptor activity and downstream signaling. Using an NF-κB-driven GFP reporter cell line that lacks endogenous TLR3, we performed "addback" experiments in which wild-type or mutant chTLR3 was reintroduced. Cells were then stimulated with the synthetic dsRNA analog polyinosinic:polycytidylic acid (poly I:C) to assess NF-κB signaling. These experiments allow us to evaluate how individual chTLR3 variants influence dsRNA sensing and signaling capacity. Understanding species-specific differences in TLR3 signaling not only provides insight into fundamental immune mechanisms but also has implications for improving poultry health and controlling viral outbreaks. These findings serve as a basis for future studies aimed at defining how TLR3 variation contributes to disease susceptibility and innate immune regulation in birds.
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M.S.