Toll Like Receptor 7 Gene PCR-SSCP Polymorphism And Its Association With Infected Pulmonary Adenomatosis In Sheep

Toll like receptors are a class of pattern recognition receptors. Now we know that TLRs can not only activate the innate immunity, but also initiate acquired immunity. The TLR7 is located inside the cell, which exists widely in many immune cells, and play an important role in the immune system, and be related to disease resistant infections, susceptibility, and immune response.In the study, we analyzed the TLR7 gene of the Suffolk and Kazakh sheep through PCR-SSCP polymorphism analysis, explored the relationship with TLR7 gene mutation and susceptibility of OPA, discussed the immune response in artificially infected with sheep pulmonary adenomatosis virus in sheep in different genotypes of TLR7 gene.The study of the relationship TLR7 gene polymorphism and the immune response of OPA infection can provide a reference for the research about breeding sheep. Artificial infection OPA can also provide an animal model for human lung cancer,and provide evidence for studying better human Lung cancer.1 Association analysis of Suffolk sheep and Kazakstan sheep Toll like receptor 7 gene SSCP polymorphism analysis of sheep pulmonary adenomatosisUsing PCR-SSCP method to analyze the polymorphism of Suffolk sheep and Kazakstan sheep TLR7 gene. Analysis showed that the TLR7 gene is polymorphic. The extracellular domain of Suffolk, ECD-2 ECD-5 ECD-6,ECD-7 have the polymorphism. ECD-2 and ECD-7 in Kazak sheep by sequencing have the polymorphism. Mutations found in Suffolk sheep with 5 point mutations, including 3 missense mutations, 2 synonymous mutations. While Kazak sheep only 3 mutations, including 2 missense mutations and 1 synonymous mutations. ECD-2 SSCP analysis showed that the Suffolk District in Hardy Weinberg unequilibrium, and the mutation with type BB, type AB,which positive frequency of endogenous Jaagsiekte sheep retrovirus infection higher than normal typeAA, these genotypes after sequencing found that 343 and 350 nucleotide mutations, the positive rate of the two mutants at the same time,infected with endogenous sheep are higher than that of the negative sheep. Mutations in A343 G, A350 G may be related to the susceptibility of the virus.2 Different types of TLR7 ECD-2 gene on the immune response in early infection of sheep pulmonary adenomatosisMutation of TLR7 nucleotide A343 G and A350 G sheep and non-mutation sheep were artificially infected with sheep pulmonary adenomatosis virus.blood tests carried out after the second day of artical infection,the inflammatory cell number reached the highest value in vivo. The mutation group result of ELISA for IFN-γ, IL-1, TNF-α, MyD88, TLR7 in serum the expression level are higher than the the expression level of non mutation group. While analysis of variance found that the differences were not significant(P > 0.05).The results suggest that the flammatory response in the mutant group have more proinflammatory factors involved in the resistance to the virus, and the immune response is more active.3 ECD-2 mutation of TLR7 gene and clinical pathological observation of sheep lung adenomaAfter 5 months, we progressed necropsy observations and tissue observation for mutation and non mutation groups The results showed that mutation group sheep had OPA symptoms, there are many gray white nodules on the lung surface. Through slice observation,we could find lung diseased sheep have different sizes of adenoma lesions and papillary hyperplasia of alveolar epithelial cells, The alveolar cells became cuboidal or low columnar cells, due to pulmonary tumor nodules compress adjacent alveolar or expansion of the adjacent alveolar fusion. Adenoma lesions with varying amounts of non adenoma alveolar group, the alveolar cavity filled with macrophages.The terminal bronchioles in columnar epithelial hyperplasia and protruding into the cavity and in the surrounding lymphocytes, plasma cells, macrophages around. OPA symptoms of mutation group are worse than that of non mutant group. ecd-2 mutant sheep of OPA are more likely to infection.In summary, TLR7 gene existed polymorphism, this polymorphism is the foundation of screening genetic markers related to disease resistance or susceptibility. In this article, we progress PCR-SSCP analysis and gene mutation frequency analysis to research the association of toll like receptor 7 gene polymorphism in sheep and sheep pulmonary adenomatosis. In order to study the relationship between TLR7 gene and disease resistance, and to provide a genetic basis for the selection of excellent gene.