Duck And Goose PRRs Clone, Analysis, Distirbutions Polymorphism And Response To Special Ligands

PRRs are conserved immune receptors which classified according to itslocalization into Membrane-bound PRRs located on the cell membrane andCytoplasmic PRRs and Secreted PRRs which can locate freely without associationwith the cells. They activate innate immune responses to protecting the host frominfection, by identifying some PAMPs. Toll like receptor are membranous receptorexcept TLR-3/7/9consider endoplasmic receptor, while NLRs and RLRs belong tocytoplasmic proteins group that regulate inflammatory and apoptotic responses. TheNLRs family has been reported in many vertebrate species where20membersreported till now and placed in two main groups according to its structure domains,NLRC group and NLRP group. NLRCs induce signals through NF-κB and MAPkinases pathway by the RIP2which is serine-threonine kinase. From the NRLPssubfamilies there are14members cloned in humans till now (NALP1to NALP14).RLRs responsible for intracellular recognition of viral RNA, where its signal inducedthrough recruit factors in N-terminal CARD domains to activate antiviral geneprograms. Five members of RLRs family have been cloned and identified invertebrates, including RIG-I and MDA5which activate antiviral signaling, whileLGP2reported to be dominant-negative inhibitor, and Dicer-I and eIF4A newlyidentified members. The avian immune system structure and function are not wellstudied like mammalian models and the majority of it based on chicken as a universalavian model, while recent researches showed that a lot of factor not common betweenchicken and other birds and there are many disease and microbes affect chicken andcause a sever effect where other birds resist it, like avian flu, in duck just apparentinfection while it lethal for chicken. Our work focused on duck as a model forAnseriformes, where duck showed different immune response to many microbes ifcompared to chicken, our work focused on identify the duck PRRs families and checkeither it functional genes or nonfunctional genes, then make analysis to its proteinstructures and trying to identifying the sites of mutation between duck and chicken asa universal bird model, so we can get more clear data about bird evolution, thentrying to study the effect of this mutant sites on immune response. We cloned andexpressed almost the complete duck TLRs family which reported as dTLR1-1/1-2/2- 2/3/4/5/7/15/21sequences in various duck tissues, also goose toll like receptor4andgoose toll like receptor5as a complete sequence and goose toll like receptor2type1and goose toll like receptor2type2partially, the phylogenic analysis showed that ourcloned genes are orthologs to the known TLRs. We reported a degree of geneticvariation in TLRs genes among the three different aves families (Anseriformes,Galliformes and Passeriformes); we also found patterns of positive selection acting onspecific amino acid sites that could be linked to species-specific differences inpathogen-associated molecular pattern recognition. We also success to clone ducknode like receptors include NOD1, NLR3, NLRX1(according to our knowledge it'sthe first actual report, not just prediction, for those NLR members in all birds till now)and NLR5and from RLRs family we cloned LGP2, EIF4A3, and Dicer I type3. Andduck CLEC16A we also reported the in-vivo and in-vitro response of duck toll likereceptor to stimulation with LPS on peritoneal macrophage as and our result clearedits completely functional receptor and consider ortholog to the TLR4. This studyprovides evidence for the evolutionary patterns and implications of TLRpolymorphism in new avian models species and extends the list of available avianimmunogenetic genes.