Simultaneous RNA-seq based transcriptional profiling of intracellular Brucella abortus and B. abortus-infected murine macrophages

• Published in: Microbial pathogenesis 2017-12, Vol.113, p.57-67
• Main Authors: Hop, Huynh Tan, Arayan, Lauren Togonon, Reyes, Alisha Wehdnesday Bernardo, Huy, Tran Xuan Ngoc, Min, WonGi, Lee, Hu Jang, Son, Jee Soo, Kim, Suk
• Format: Article
• Language: English
• Subjects: Brucella abortus; Host defenses; Infected BMM; RNA isolation; RNA-seq; Virulence factors
• ISSN: 0882-4010; 1096-1208
• DOI: 10.1016/j.micpath.2017.10.029

Brucella is a zoonotic pathogen that survives within macrophages; however the replicative mechanisms involved are not fully understood. We describe the isolation of sufficient Brucella abortus RNA from primary host cell environment using modified reported methods for RNA-seq analysis, and simultaneously characterize the transcriptional profiles of intracellular B. abortus and bone marrow-derived macrophages (BMM) from BALB/c mice at 24 h (replicative phase) post-infection. Our results revealed that 25.12% (801/3190) and 16.16% (515/3190) of the total B. abortus genes were up-regulated and down-regulated at >2-fold, respectively as compared to the free-living B. abortus. Among >5-fold differentially expressed genes, the up-regulated genes are mostly involved in DNA, RNA manipulations as well as protein biosynthesis and secretion while the down-regulated genes are mainly involved in energy production and metabolism. On the other hand, the host responses during B. abortus infection revealed that 14.01% (6071/43,346) of BMM genes were reproducibly transcribed at >5-fold during infection. Transcription of cytokines, chemokines and transcriptional factors, such as tumor necrosis factor (Tnf), interleukin-1α (Il1α), interleukin-1β (Il1β), interleukin-6 (Il6), interleukin-12 (Il12), chemokine C-X-C motif (CXCL) family, nuclear factor kappa B (Nf-κb), signal transducer and activator of transcription 1 (Stat1), that may contribute to host defense were markedly induced while transcription of various genes involved in cell proliferation and metabolism were suppressed upon B. abortus infection. In conclusion, these data suggest that Brucella modulates gene expression in hostile intracellular environment while simultaneously alters the host pathways that may lead to the pathogen's intracellular survival and infection. •The transcriptional profiles of B. abortus and BMM were analyzed by RNA-seq approach.•The up-regulated B. abortus genes are involved in DNA, RNA manipulations and protein biosynthesis.•The down-regulated B. abortus genes are involved in energy production and metabolism.•The host defense genes are induced but cell proliferation and metabolism genes are suppressed in B. abortus infected BMM.