The Study Of TRIM5Polymorphism Of Cynomolgus Macaque (Macaca Fascicularis) In Captivity In China

Acquired Immune Deficiency Syndrome (AIDS) has become one of the most seriousproblems to our human in the21stcentury, which is a serious threat to human health, causinghuge economic losses and serious social problems. Non-human primate AIDS disease animalmodels are essential for better understanding of the pathogenesis of AIDS and develop drugs&vaccines for its treatment. Researches show that homology of the genomes between humanand Cynomolgus matched highly, and the Cynomolgus macaque (Macaca fascicularis)disease model suitably simulates the onset and progression of human disease. What is more,Cynomolgus monkey resources are extremely rich, and a number of Cynomolgus monkey arefarmed in China. Therefore, the Cynomolgus monkey AIDS model is set up to study AIDSbetter. The TRIM5α restriction factor can protect some species of monkeys from HIVinfection, but the retroviral restriction factor TRIMCyp derived from the TRIM5gene in oldworld monkeys is generally considered unable to block HIV-1replication. TRIMCyp fusiongene is a novel hot spot of anti-HIV-1factor. Locations of the different species of CypAinserted into TRIM5genes are diverse, and in the old world monkey the TRIMCyp fusion geneformed by the LINE-1-mediated retro-transposition of a CypA pseudogene cDNA into3’terminal or3’-UTR of TRIM5gene in these monkeys. The regional differences, genefrequencies, genotypes, and retrovirus restrictive activities of TRIMCyp vary among differentprimate species. It was reported that TRIMCyp-major(DK) can block the activities of HIV-1,but not HIV-2; and TRIMCyp-minor(NE) can block HIV-2, but not HIV-1. Therefore, aCynomolgus monkey with TRIMCyp NE haplotype may be constructed into HIV-1animalmodel. Our research provides interesting findings that contribute towards a more firm basis offurther studies of HIV-1animal models and relevant pathogenesis.The polymorphism and characterization of Cynomolgus TRIMCyp had been studied inseveral areas of Southeast Asia, and the frequency and prevalence of Cynomolgus TRIMCypin China were studied partly. But the global existence situation of Cynomolgus in captivity inChina has not been clearly elucidated. Cynomolgus monkeys are the most nonhuman primatesbred in China, and those with TRIMCyp might be used as non-human primate model of HIV.On the basis of abundant resources of Cynomolgus in China, Our study investigated the polymorphisms and frequencies of TRIMCyp of total1594Cynomolgus macaques sampledfrom11feedlots in5provinces in this study. The results are as follows:(1) The location of CypA inserted into TRIM5gene by the LINE-1-mediated retrotrans-position of a CypA pseudogene cDNA into3’ terminal or3’-UTR of TRIM5gene in the oldworld monkey is evaluated from previous reports. Then, the screened primers of TRIMCypwere designed. The TRIMCyp fusion gene was screened using the PCR. The results showedthat the average allele frequencies of TRIMCyp and TRIM5were13.36%and86.64%respectively. The most is the form of TRIM5/TRIMCyp heterozygous (87.60%), and TRIMCypfusion gene of Cynomolgus monkey from each field is almost the same condition, only a littledifference (7.65%~19.79%). But the result of TRIM5/TRIMCyp heterozygote of differentcountries or areas in Southeast Asia is53.16%and the TRIMCyp frequencies as high as100%(Philippines), and it is far higher than the average of13.36%in China. We speculate thatpotentially the latter were isolated groups established since1978. In addition, by thecomparison and analysis of CypA sequencing results, we evaluate the distribution ofTRIMCyp-major(DK) and TRIMCyp-minor(NE). The results showed that the TRIMCyp-major(DK) accounted for95.07%; Whereas the TRIMCyp-minor(NE) of Cynomolgusmonkey was very few, and the TRIMCyp-minor(NE) frequency is only4.93%, and only onefor the TRIMCyp homozygous(NE), significantly lower than the TRIMCyp-minor (NE)frequency of Cynomolgus monkey of three countries in southeast Asia(11.1%~14.3%).TRIMCyp-minor(NE) can not block HIV-1, and human AIDS disease is usually caused by theretrovirus HIV-1, thus a Cynomolgus monkey with TRIMCyp NE haplotype may beconstructed into HIV-1animal model. It is likely to build more Cynomolgus monkey withTRIMCyp homozygous(NE) by establishing a closed group of monkey, and it can promotefurther studies of HIV-1animal models and relevant pathogenesis.(2) We screened the polymorphism of TRIMCyp in a small group of TRIMCypCynomolgus monkey, and the results showed that SNPs which were complete linkage withTRIM5. CypA insertion were identified only in TRIM5exon7and8of Cynomolgus macaques.Then they were identified in individuals of Cynomolgus macaques, Chinese rhesus macaques,pig-tailed macaques and Macaca thibetana. Remarkably, eight novel SNPs in exon7andexon8which were complete linkage with TRIM5. CypA insertion were identified, two of them in exon8trended to form a predicted ESS (Exonic Splicing Silencers) which mightexplain why exons8was100%absent in all TRIMCyp transcripts since there was no anymutation at the5’ splice donor (GT) and3’ splice acceptor (AG) sites of intron7. Overall, theresults of the present study will help understand the genetic background of TRIMCyp ofCynomolgus macaque in captivity in China and the possible reason of exon8absent in allTRIMCyp transcripts.