Distribution of interleukin 17 receptor mRNA in rhesus macaques / 中国免疫学杂志

Objective:To delineate the distribution and abundance of interleukin 17 receptor mRNA in normal tissues of rhesus macaques.Methods:Tissue samples were collected from different systems including lymphoid system,digestive system and genital system and the mRNA levels of IL-17RA and IL-17RC were examined by real time RT-PCR.Meanwhile the levels of IL-17RA and IL-17RC mRNA in the PBMCs of rhesus macaques,African green monkeys and cynomolgus monkeys were examined.Results:IL-17RA mRNA was expressed in all the tested tissues and the levels were significantly different among these systems (P＜0.05).IL-17RC mRNA level in the large intestine was higher than that in the small intestine.IL-17RA mRNA level in rhesus macaque PBMCs was 3 and 5 times higher than that of cynomolgus monkeys and African green monkeys respectively,while IL-17RC mRNA in the PBMCs was below the limit of detection.Conclusion:IL-17RA is widely distributed in rhesus macaques;the expression level is different among different tissues and species.

Construction and characterization of a new simian/human immunodeficiency viruses clone carrying an env gene derived from a CRF07_BC strain / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque.</p><p><b>METHODS</b>A SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion.</p><p><b>RESULTS</b>One SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection.</p><p><b>CONCLUSIONS</b>We conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.</p>

Disease progression patterns of SHIV-KB9 in rhesus macaques of Chinese origin in comparison with Indian macaques / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To develop a model of SHIV-KB9/Chinese origin rhesus (Ch Rh) macaques for vaccine research and to compare the pathogenesis of SHIV-KB9 in Ch Rh macaques with that reported in Indian rhesus (Ind Rh) macaques.</p><p><b>METHODS</b>Seven mamu-A*01 negative Ch Rh macaques were inoculated intravenously with 1-10000 MID50 of SHIV-KB9. The monkeys were monitored for viral load, CD4, CD8, SHIV-specific antibody and virus genetic variation. The results were compared with those previously observed in Ind Rh macaques.</p><p><b>RESULTS</b>As compared to that observed in Ind Rh macaques, SHIV-KB9 in Ch Rh macaques displayed three identical disease progression patterns. However, the primary pattern was not identical between the two subspecies. The level of plasma viremia differed in SHIV-KB9-infected Ch Rh macaques which exhibited different outcomes from those in Ind Rh macaques. Generally, the values of viral load and the maintenance of CD4+ T cells were associated with humoral responses. Otherwise, the viral genetic distances (divergence, diversity) were larger in animals (M419, M425) with their CD4+ T cells profoundly depleted.</p><p><b>CONCLUSION</b>The model of SHIV-KB9/Ch Rh macaques displays a relatively slow progression to AIDS compared with Ind Rh macaques, which may more accurately reflect the potential of candidate vaccines in humans.</p>

Establishment of a real-time RT-PCR to detect plasma viral load of simian/human immunodeficiency virus CN97001 during its in vivo passage in rhesus monkeys / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a real-time RT-PCR based plasma virus quantification method and monitor the plasma viral load of SHIV-CN97001 during its in vivo passages in rhesus macaques.</p><p><b>METHODS</b>Viral RNA standards were prepared by in vitro transcription and one-tube real-time RT-PCR were established and optimized using TaqMan EZ RT-PCR CORE REAGENTS and TaqMan probes and primers directed to the 91 bases within the conserved gag region of SHIV. Plasma viral RNA of 126 plasma samples from rhesus macaques of different viral passages was quantified.</p><p><b>RESULTS</b>The PCR system was optimized by using serial dilution of standards, and the viral RNA load was detected. The lowest limit of the standard curve reached 2x10(-2) copies/ml. The correlation (r>0.99) and the repetition (CV=4.14 percent) also met the requirement. It was revealed that the viral RNA load of third passage was the highest. Generally, the viral load peaks (10(5)-10(6) copies/ml) appeared at the fourteenth day after the infection or inoculation.</p><p><b>CONCLUSION</b>The method of one-tube real-time RT-PCR was established successfully, which may provide a sensitive way to qualify SHIV viral load. This will contribute to the establishment and application of SHIV/rhesus macaque models. It was also found that the replicative ability of SHIV-CN97001 was enhanced during the first 2 in vivo passages.</p>