Micro RNA in Exosomes from HIV-Infected Macrophages.
Micro RNA in Exosomes from HIV-Infected Macrophages.
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来自HIV感染的巨噬细胞的外泌体中的微RNA。
DOI：
10.3390/ijerph13010032
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发表时间：
2015-12-22
影响因子：
--
通讯作者：
Bond VC
中科院分区：
文献类型：
--
作者：
Roth WW;Huang MB;Addae Konadu K;Powell MD;Bond VC
外切体是由细胞分泌的小的膜结合的囊泡，其功能是在细胞之间运送RNA和蛋白质。为了研究外体微小RNA(MiRNA)在HIV-1感染早期阶段的作用，我们比较了HIV感染的巨噬细胞和未感染的巨噬细胞外体中的miRNA。来自未感染供者的原代人类单核细胞被分化为巨噬细胞(MDM)，后者要么被模拟感染，要么被嗜巨噬细胞的HIV-1bal株感染。从培养上清液中回收外切体，用碘二醇密度梯度离心法从病毒颗粒中分离出外切体。从纯化的外切体中制备低相对分子质量的RNA组分。预扩增后，将RNA与含有1200种已知和未知功能miRNA的探针的微阵列杂交。我们在感染和未感染的MDM外体中观察到48种miRNA。此外，38个miRNAs存在于感染细胞外体中，但不存在于未感染细胞外体中。其中，13个miRNAs在HIV感染细胞的外切体中上调，其中4个miRNA物种的表达增加了10倍以上。虽然已在HIV感染细胞中鉴定出许多miRNA物种，但对这些细胞外体中miRNA的含量知之甚少。在未来，我们计划调查我们确定的上调的miRNA物种是否在HIV-1阳性患者的外切体中增加。
Exosomes are small membrane-bound vesicles secreted by cells that function to shuttle RNA and proteins between cells. To examine the role of exosomal micro RNA (miRNA) during the early stage of HIV-1 infection we characterized miRNA in exosomes from HIV-infected macrophages, compared with exosomes from non-infected macrophages. Primary human monocytes from uninfected donors were differentiated to macrophages (MDM) which were either mock-infected or infected with the macrophage-tropic HIV-1 BaL strain. Exosomes were recovered from culture media and separated from virus particles by centrifugation on iodixanol density gradients. The low molecular weight RNA fraction was prepared from purified exosomes. After pre-amplification, RNA was hybridized to microarrays containing probes for 1200 miRNA species of known and unknown function. We observed 48 miRNA species in both infected and uninfected MDM exosomes. Additionally, 38 miRNAs were present in infected-cell exosomes but not uninfected-cell exosomes. Of these, 13 miRNAs were upregulated in exosomes from HIV-infected cells, including 4 miRNA species that were increased by more than 10-fold. Though numerous miRNA species have been identified in HIV-infected cells, relatively little is known about miRNA content in exosomes from these cells. In the future, we plan to investigate whether the upregulated miRNA species we identified are increased in exosomes from HIV-1-positive patients.