title: Simultaneous transcriptional profiling of bacteria and their host cells.
abstract: We developed an RNA-Seq-based method to simultaneously capture prokaryotic and eukaryotic expression profiles of cells infected with intracellular bacteria. As proof of principle, this method was applied to Chlamydia trachomatis-infected epithelial cell monolayers in vitro, successfully obtaining transcriptomes of both C. trachomatis and the host cells at 1 and 24 hours post-infection. Chlamydiae are obligate intracellular bacterial pathogens that cause a range of mammalian diseases. In humans chlamydiae are responsible for the most common sexually transmitted bacterial infections and trachoma (infectious blindness). Disease arises by adverse host inflammatory reactions that induce tissue damage & scarring. However, little is known about the mechanisms underlying these outcomes. Chlamydia are genetically intractable as replication outside of the host cell is not yet possible and there are no practical tools for routine genetic manipulation, making genome-scale approaches critical. The early timeframe of infection is poorly understood and the host transcriptional response to chlamydial infection is not well defined. Our simultaneous RNA-Seq method was applied to a simplified in vitro model of chlamydial infection. We discovered a possible chlamydial strategy for early iron acquisition, putative immune dampening effects of chlamydial infection on the host cell, and present a hypothesis for Chlamydia-induced fibrotic scarring through runaway positive feedback loops. In general, simultaneous RNA-Seq helps to reveal the complex interplay between invading bacterial pathogens and their host mammalian cells and is immediately applicable to any bacteria/host cell interaction.
authors: Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.
authors: Michael S
authors: MS
authors: Humphrys
authors: Todd
authors: T
authors: Creasy
authors: Yezhou
authors: Y
authors: Sun
authors: Amol C
authors: AC
authors: Shetty
authors: Marcus C
authors: MC
authors: Chibucos
authors: Elliott F
authors: EF
authors: Drabek
authors: Claire M
authors: CM
authors: Fraser
authors: Umar
authors: U
authors: Farooq
authors: Naomi
authors: N
authors: Sengamalay
authors: Sandy
authors: S
authors: Ott
authors: Huizhong
authors: H
authors: Shou
authors: Patrik M
authors: PM
authors: Bavoil
authors: Anup
authors: A
authors: Mahurkar
authors: Garry S A
authors: GS
authors: Myers
mesh_terms: D045744:Cell Line, Tumor; D002692:Chlamydia trachomatis / Q000235:genetics* / Q000378:metabolism; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism* / Q000382:microbiology; D025461:Feedback, Physiological; D020869:Gene Expression Profiling; D054884:Host-Pathogen Interactions; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D054892:Metagenome; D017423:Sequence Analysis, RNA; D019063:Tenascin / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome
chemical_list: C486399:GREM1 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D019063:Tenascin; D003094:Collagen