Relationship Analysis On Different Alleles Of Major Histocompatibility Complex Class Ⅱ And SIV/HIV-2Virus Load Of Cynomolgus Macaques

MHC is an important factor in the determination of susceptibility and developing speed ofdisease, and it is one of the key factors in the inhibition of viral replication on the aspect ofhumoral immunity, cellular immunity and innate immunity. The unclear geneticbackground of animals plays a great limitation role in the breakthrough of importantdiseases. Following the completion of whole genome sequencing, crab-eating macaque hasbeen an very important experimental animal. At present, an more importantapplication direction of cynomolgus monkey is as an animal model in HIV-1pathological study. Researchers didn’t understand the immune reaction mechanism whenhuman are infected with HIV, but experimental data provided by non-human primate is theonly usefully exploratory data to verify the immune protection function of human. TheMHC gene polymorphism of cynomolgus monkey can significantly effect the results ofdrug experiment, and was connected with viral disease. The chanllege of SIV/HIV-2ofdifferent macaque species and macaques from different regions is various. On theprimary research basis, my artical also make use of the cynomolgus monkey groups toidentify the polymorphism of three MHC II gene (Mafa-DPA, DQA, DRA) and takeadvantage of SIV/HIV-2infection experiments to make a comprehensive analyze on MHCgenetic background and co-occurring mode analysis in groups, and anylyze the relationshipof MHC alleles and SIV/HIV-2virus load to understand the role of different alleles inSIV/HIV-2infection. The results of our study are as follows:(1) The identification of the polymorphism of MHC class II alleles:We adopt the method of TA clone sequencing at transcriptional level to identify18DPAalleles from the cynomolgus monkey group which consists of23individuals, including1high-frequency alleles (DPA-C7-2, corresponding the frequency of23.9%). Similarly, weidentified22DQA alleles from the same group. We also identified8DRA alleles from thesame group, including1high-frequency alleles (DRA-C7-6, corresponding the frequencyof41.3%). By the co-occurring analysis of Mafa-DPA, DQA, DRA alleles, in the groupwhich consists of23individuals, we found the alleles combination ofDPA-C7-2-DQA-C7-8-1, DRA-C30-12-DQA-C7-8-1, DRA-C7-3-DRA-C7-5, DRA-C7-5-DPA-C7-2, DRA-C30-12-DPA-C7-2were identified in19.2%,15.4%,11.5%,11.5%,11.5%individuals. This suggests that MHC II alleles sharing is a major characteristic ofcrab-eating macaques. (2) The relationship analysis on different MHC II alleles and SIV/HIV-2viral load:As for the two alleles of DPA, the results of independent sample t test are as follows: whenPBMC were infected with SIV for5days, t=1.005, P=0.419>0.05, it suggests that twoalleles made no significant difference in the SIVmac239viral load; when infected with SIVfor7days, t=0.033, P=0.975>0.05, it suggests that two alleles made no significantdifference in the SIVmac239viral load; when infected with HIV-2for5days, t=-0.616, P=0.549>0.05, it suggests that two alleles made no significant difference in the HIV-2RODviral load; when infected with SIV for7days, t=-0.789, P=0.446>0.05, it suggests thattwo alleles made no significant difference in the HIV-2ROD viral load. As for the twodifferent alleles DQA-C6-1-1and DQA-C7-4-1of DQA, the results of independent samplet test are as follows: when PBMC were infected with SIV for5days, t=-0.516, P=0.618>0.05, it suggests that two alleles made no significant difference in the SIVmac239viralload; when infected with SIV for7days, t=-0.888, P=0.400>0.05, it suggests that twoalleles made no significant difference in the SIVmac239viral load; when infected withHIV-2for5days, t=1.531, P=0.160>0.05, it suggests that two alleles made nosignificant difference in the HIV-2ROD viral load; when infected with SIV for7days, t=0.557, P=0.593>0.05, it suggests that two alleles made no significant difference in theHIV-2ROD viral load. As for the allele DRA-C7-6and two allele combinationDRA-C30-12-DRA-C7-6and DRA-C7-5-DRA-C7-6of DQA, the results of one-wayanalysis of variance are as follows: when PBMC were infected with SIV for5days, F=0.129, P=0.880>0.05, it suggests that allele DRA-C7-6and their two combinations madeno significant difference in the SIVmac239viral load; when infected with SIV for7days,F=0.777, P=0.480>0.05, it suggests that allele DRA-C7-6and their two combinationsmade no significant difference in the SIVmac239viral load; when infected with HIV-2for5days, F=1.196, P=0.332>0.05, it suggests that allele DRA-C7-6and their twocombinations made no significant difference in the SIVmac239viral load; when infectedwith SIV for7days, F=0.925, P=0.419>0.05, it suggests that allele DRA-C7-6andtheir two combinations made no significant difference in the SIVmac239viral load. LSDand Duncan were used to do the multiple comparision of DRA-C30-12-DRA-C7-6,DRA-C7-5-DRA-C7-6and DRA-C7-6, the results shown that DRA-C7-6and DRA-C30-12-DRA-C7-6, DRA-C7-6and DRA-C7-5-DRA-C7-6, DRA-C30-12-DRA-C7-6andDRA-C7-5-DRA-C7-6, each pair of them made no significant difference in the SIVmac239/HIV-2ROD viral load.