Exosomal RNA from Mycobacterium tuberculosis‐Infected Cells Is Functional in Recipient Macrophages

Exosomes are extracellular vesicles released by cells that carry proteins, lipids and nucleic acids and function in intercellular communication. Previously, we determined that exosomes released from Mycobacterium tuberculosis (M.tb)-infected macrophages carry mycobacterial proteins and lipids. However, the RNA composition within these exosomes has not been defined. In this study, we characterized the exosomes released from M.tb-infected macrophages and identified a cohort of mouse messenger RNA (mRNA) and microRNA (miRNA). Quantitative reverse-transcriptase polymerase chain reaction analysis showed less abundance of miRNAs in exosomes released from infected compared with uninfected macrophages. Moreover, more than 100 transcripts were found to be enriched or unique to exosomes from infected cells including transcripts involved in regulating an immune response. The exosomal RNA could be transferred and expressed in naïve macrophages and was biologically active, stimulating production of inflammatory mediators and inducing apoptosis in recipient cells. Interestingly, we also identified mycobacterial transcripts in exosomes released from infected macrophages. To our knowledge, this is the first study to identify bacterial-derived RNA in exosomes. Our results suggest that exosomal RNA released from M.tb-infected macrophages may have functional and diagnostic potential in the context of a mycobacterial infection.