An Association Study Of TRIM5α Alleles With HIV-2_ROD And SIVmac239 Infection In Cynomolgus And Chinese Rhesus Macaques

TRIM5α is an important factor in the determination of susceptibility and developing speed of disease, and it is one of the key factors in the inhibition of viral replication on the aspect of innate immunity, humoral immunity, and cellular immunity. Previous studies in Indian-origin rhesus macaques indicated that TRIM5α polymorphism affected retrovirus replication. Chinese-origin rhesus macaques(CR) and cynomolgus macaques(CE) have become very important medical experimental animals and increasingly used in HIV/AIDS research because Indian-origin rhesus macaques was limited to export. So far the effects of TRIM5α polymorphism on viral replication in Chinese-origin rhesus macaques are still unclear. Meanwhile, no research was carried out to study the effect in cynomolgus macaques of Vietnamese origin(CM). In addition, we previously identified a 6-bp deletion in the TRIM5α gene in the 39 cynomolgus macaques of Vietnamese origin that results in the loss of two amino acids(Thr339 and Phe340) at the B30.2(SPRY) domains. A high frequency(97.5%) of this mutation was detected in the cynomolgus macaque population, indicating that this deletion has become virtually fixed in the cynomolgus macaque of Vietnamese origin. The next question, then, are how do the cynomolgus macaque population cope with selection pressure from HIV/SIV during evolution since the loss of two critical functionality amino acids in their TRIM5α? And whether there are any other TRIM5α functional sites in response to these retrovirus infections? To answer these questions, on the primary research basis, we recruited 40 unrelated cynomolgus macaques of Vietnamese origin and 68 unrelated Chinese-origin rhesus macaques to identify the polymorphism of TRIM5α and to analysis the TRIM5α genetic background of these two macaques. We also study the effect of TRIM5α polymorphism on HIV-2ROD and SIVmac239 replication among peripheral blood mononuclear rcells(PBMC), to clarify TRIM5 alpha functional domains in the role of viral replication and the possible mechanism in the two macaques. The results of our study are as follows:(1) The identification of the polymorphism of TRIM5α allelesWe identify 20 SNPs in TRIM5α exon 4 and exon 8 from the cynomolgus macaques of Vietnamese origin and Chinese rhesus macaques groups which consists of 40 individuals and 68 individuals, respectively. our results revealed the presence of 16 SNPs which form 10 haplotypes and 14 alleles, including 1 high-frequency haplotype(corresponding the frequency of 61.25%), a species-specific alleles with a low frequency, and a 1 high-frequency alleles(corresponding the frequency of 40%) in cynomolgus macques. We identified a 6-bp deletion in the TRIM5α gene in all of the cynomolgus macaque of Vietnamese origin that results in the loss of two amino acids(Thr339 and Phe340) at the B30.2(SPRY) domains. This is in line with previous research that a high frequency(97.5%) of this mutation was detected in 39 cynomolgus macaque population, indicating that this deletion has become virtually fixed in the cynomolgus macaque of Vietnamese origin.Similarly, we identified 9 SNPs which form 9 haplotypes and 24 alleles, oniy 1 high-frequency haplotype(corresponding the frequency of 28.68%) was identified in Chinese rhesus macaques. We also find that all of the SNPs identified in the B30.2(SPRY) domains formed a haplotype in Chinese rhesus macaques.(2) The relationship analysis on different TRIM5α alleles and HIV-2/SIV infectionAs a result, we found that 3 variants, A333S/TFP339-341ΔΔQ/S422 L, located in B30.2(SPRY) domain of Chinese rhesus macaques, formed a haplotype and significantly affected HIV-2ROD replication. In addition, Y178 H located in Coiled-coil domain, which changed TRIMCyp expression and stability, also affect cynomolgus macaque TRIM5α-mediated HIV-2ROD restriction. Moreover, two variants, E222 K and E236 D, located in Coiled-coil domain of Chinese rhesus macaques, exhibited to be able to alter anti-SIVmac239 activity. However, none of variants was correlated with cynomolgus macaque TRIM5α-mediated SIVmac239 restriction. Based on these results, we concluded that CM TRIM5α polymorphism did not affect SIVmac239 replication, but could alter HIV-2ROD infection as well as different domain of CR TRIM5α was responsible for restricting different virus replication.