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构建并鉴定一株携带 CRF07_BC 株来源的 env 基因的新型猴/人免疫缺陷病毒克隆。

Construction and characterization of a new simian/human immunodeficiency viruses clone carrying an env gene derived from a CRF07_BC strain.

机构信息

College of Lifesciences, Nankai University, Tianjin, China.

出版信息

Chin Med J (Engl). 2009 Dec 5;122(23):2874-9.

Abstract

BACKGROUND

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.

METHODS

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.

RESULTS

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.

CONCLUSIONS

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.

摘要

背景

CRF07_BC 重组毒株一直是中国最主要流行的 HIV-1 毒株之一,因此有必要且迫切需要开发针对 HIV-1 CRF07_BC 的相关动物模型来评估候选疫苗。含有能够感染中国恒河猴的中国 CRF07_BC HIV-1 env 基因的高复制能力的猴免疫缺陷病毒(SHIV)构建体将成为 HIV 疫苗开发的重要工具。本研究的目的是研究是否可以使用含有中国 HIV-1 分离株病毒 env 片段的 SHIV XJDC6431 感染人外周血单核细胞(PBMC),并在中国恒河猴中建立感染。

方法

通过用来自 HIV-1 CRF07_BC 株的相应片段替换 SHIV KB9 的 rev/env 基因来构建 SHIV 株。在来自非人类灵长类动物和人类的 PBMC 中体外测定 SHIV 克隆的感染活性。最后,通过静脉输注将 1 只中国恒河猴(Macaca mulatta)感染 1 种 SHIV。

结果

生成了一种命名为 SHIV XJDC6431 的 SHIV 克隆,它可以感染猕猴和人 PBMC。该克隆产生的病毒也可以有效地感染 CCR5 表达的 GHOST 细胞系,表明它使用 CCR5 作为其核心受体。最后,将病毒静脉接种到 1 只中国恒河猴体内。最终,动物感染,感染后 3 天内出现病毒血症。感染急性期病毒载量达到每毫升血浆 105 拷贝的病毒 RNA,感染后持续 10 周。

结论

我们得出结论,SHIV XJDC6431 是一种 R5 嗜性嵌合病毒,不仅可以在体外,而且可以在体内感染中国恒河猴。虽然用 SHIV XJDC6431 接种的动物没有出现病理表型,但病毒在血浆中仍以持续的病毒载量有效复制。这表明,该 SHIV 可用于测试针对中国 HIV-1 CRF_07BC 重组株的候选 AIDS 疫苗。

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