The Research On Polymorphism Of Trim5α And Major Histocompatibility Complex Class Ⅱ Of Macaca Mulatta

The host restriction innate immune molecules TRIM5and MHC is important factorswhich determine disease susceptibility and pathogenesis speed in human genes. One of thefactors is that play a crucial role in inhibition of viral replication in the various levels of theimmune response. Researching TRIM5gene polymorphism with HIV-1infection anddisease progression correlation has important theoretical and practical value.Many studieshave shown that the cynomolgus monkey MHC gene polymorphism significantly affectdrug experimental results, but also affect the SIV virus infection and replication.Cynomolgus monkeys has been widely used in medical and pharmaceutical research,but theresearch on cynomolgus monkey MHC molecule gene polymorphisms is still verylimited,The genetic background on MHC is not clear is one of insurmountable obstacleswhich greatly limit our ability to achieve a breakthrough in the study of the majordiseasesprogress.studying MHC gene polymorphism has a very important significance. Inthis study,utilizing cynomolgus monkeys from Vietnamese origin and Chinese rhesusmacaques, PCR products from all eight exons of TRIM5were directly sequenced from theDNA level,respectively. and further analysed DNA sequence polymorphism including the5’-UTR region, B30.2(SPRY) and coiled-coil functional domain by using biologysoftware.provided important message for expression and regulation of mRNA for the geneand in-depth study the interaction between the coiled-coil and B30.2(SPRY). Inaddition,this study also used the cynomolgus monkey groups, continued to analyse andresearch the polymorphism on MHC-II class (DOA, DPA, DQA and DRA) gene exon2ofcynomolgus monkey on the current basis from the cDNA level. Identified MHC-II classalleles of cynomolgus monkey groups and revealed alleles distribution pattern in the groupto provide important reference to establish a clear genetic background of cynomolgusmonkeys animal models. The results are as follows,Firstly,the polymorphism researchof8exon of TRIM5α gene of Cynomolgusmonkeys and Chinese rhesus macaquesTRIM5is a retroviral restriction factor, in which the B30.2(SPRY) and coiled-coildomains cooperate to determine the specificity of TRIM5-mediated capture of retroviral capsids. Here, all exons of TRIM5were analyzed in39cynomolgus macaques (CE) and29Chinese rhesus macaques (CR). Our results revealed the presence of22alleles using thePHASE2.0software package (PHylogenetics And Sequence Evolution), including2novelspecies-specific alleles with a low frequency in CE in exons4and8, which encoded thecoiled-coil and B30.2(SPRY) domains, respectively. Nine alleles were detected in both CEand CR, while4alleles were likely shared between the species. Of these alleles, the highestfrequencies of38%and26%occurred in CE and CR, respectively. Importantly, we foundthat some alleles encoded the same coiled-coil domain, but not the SPRY domain. Incontrast, other alleles encoded the same SPRY domain, but not the coiled-coil domain. Ourfindings will contribute to the understanding of the interaction between the2domains andthe determination of the specificity of TRIM5-mediated capture of retroviral capsids. Ourresults from the phylogenetic trees constructed for CE and CR suggested that themacaques, ability to inhibit SIV replication became gradually stronger if they carriedcorresponding alleles in4clades (clade4to clade7). More interesting, in clade3, both novelallele pairs (4E100a,10E147a) and allele pairs (7R17b and13R11b), which had the strongability to inhibit SIV replication, originated from the same ancestral allele, suggesting thatthe novel alleles might play a key role to determine an animal,s ability to inhibit SIV/HIVreplication.Secondly,the research of polymorphism of exon2of MHCII genes (DOA,DPA, DQA andDRA) of Cynomolgus monkeyThe research carried out identification and polymorphism analysis of Mafa-DOA,DPAMafa-DOA,DPA,-DQA,-DRA four genes of Vietnamese origin cynomolgus monkey oncDNA level.Firstly,15DOA alleles were identified in30individuals, and all were newalleles in the research,including three high-frequency alleles gene;Secondly,26DPA alleleswere identified in28individuals, sixteen were new alleles in the research,6high-frequencyalleles gene among them.Once more,17DQA alleles were identified in30individuals,14were new alleles in the research,3high-frequency alleles gene among them. Finally,9DRA alleles were identified in30individuals,2were new alleles in the research,2high-frequency alleles gene among them.In the population in the study,the most high-frequency alleles such as Mafa-DOA*010207(33.3%) Mafa-DPA*0209(25%)Mafa-DQA1*260103(36.7%) Mafa-DRA*010201(90%),may represent the target ofimportant immune function of organism.By further analysed,the highest frequency ofco-occurring allelic genes is the combination of Mafa-DQA1*26:01:03andMafa-DRA*01:02:01(36.7%),second high frequency of co-occurring allelic genes is thecombination of Mafa-DRA*01:02:01and Mafa-DOA*01:02:07(33.3%), these instructionsthe combination of co-occurring allelic genes of Mafa-DQA/DRA, Mafa-DRA/DOA is thecharacteristics in cynomolgus macaques of Vietnamese origin in research.This study focused on the Coiled-coil and SPRY/B30.2of TRIM5, two importantfunctional domains, the exon2in DOA, DPA, DQA and DRA locus of chain of MHCII,these functional domains and identification of polymorphic loci on exon, for furtherin-depth study Trim5gene and the polymorphism on MHC and its correlation with HIVsusceptibility, laid a solid foundation for understanding the effect of each allele in AIDS,and will help to understand the pathogenesis of AIDS, is important for vaccine research,evaluation and development of drug at the same time.