Abstract
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.
Highlights
Micro RNAs are short (20–22 nt) non-coding RNA molecules which are generated from larger precursors, and typically act by binding to 3’ untranslated regions of mRNA transcripts [1].they are able to regulate gene expression at the post-transcriptional level
Exosomes Were Purified from HIV-Infected Monocyte-Derived Macrophages (MDM)
We were able to detect more than 80 different Micro RNAs (miRNA) in exosomes from primary donor macrophages infected by the macrophage-tropic HIV-1 BaL strain
Summary
Micro RNAs (miRNA) are short (20–22 nt) non-coding RNA molecules which are generated from larger precursors, and typically act by binding to 3’ untranslated regions of mRNA transcripts [1]. They are able to regulate gene expression at the post-transcriptional level. A number of studies have sought to clarify the role of miRNA in HIV disease [2,3,4]. The levels of a number of different host cell miRNA species have been reported to increase or decrease after HIV-1 infection [4,5]. Some miRNAs are thought to be involved in mediating immune suppression or in establishment of viral latency [6]
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