title: Host response during Yersinia pestis infection of human bronchial epithelial cells involves negative regulation of autophagy and suggests a modulation of survival-related and cellular growth pathways.
abstract: Yersinia pestis (Yp) causes the re-emerging disease plague, and is classified by the CDC and NIAID as a highest priority (Category A) pathogen. Currently, there is no approved human vaccine available and advances in early diagnostics and effective therapeutics are urgently needed. A deep understanding of the mechanisms of host response to Yp infection can significantly advance these three areas. We employed the Reverse Phase Protein Microarray (RPMA) technology to reveal the dynamic states of either protein level changes or phosphorylation changes associated with kinase-driven signaling pathways during host cell response to Yp infection. RPMA allowed quantitative profiling of changes in the intracellular communication network of human lung epithelial cells at different times post infection and in response to different treatment conditions, which included infection with the virulent Yp strain CO92, infection with a derivative avirulent strain CO92 (Pgm-, Pst-), treatment with heat inactivated CO92, and treatment with LPS. Responses to a total of 111 validated antibodies were profiled, leading to discovery of 12 novel protein hits. The RPMA analysis also identified several protein hits previously reported in the context of Yp infection. Furthermore, the results validated several proteins previously reported in the context of infection with other Yersinia species or implicated for potential relevance through recombinant protein and cell transfection studies. The RPMA results point to strong modulation of survival/apoptosis and cell growth pathways during early host response and also suggest a model of negative regulation of the autophagy pathway. We find significant cytoplasmic localization of p53 and reduced LC3-I to LC3-II conversion in response to Yp infection, consistent with negative regulation of autophagy. These studies allow for a deeper understanding of the pathogenesis mechanisms and the discovery of innovative approaches for prevention, early diagnosis, and treatment of plague.
authors.affiliation: National Center for Biodefense and Infectious Diseases and School of Systems Biology, George Mason University Manassas, VA, USA.
authors.forename: Farhang
authors.initials: F
authors.lastname: Alem
authors.affiliation: National Center for Biodefense and Infectious Diseases and School of Systems Biology, George Mason University Manassas, VA, USA.
authors.forename: Kuan
authors.initials: K
authors.lastname: Yao
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases Frederick, MD, USA.
authors.forename: Douglas
authors.initials: D
authors.lastname: Lane
authors.affiliation: Center for Applied Proteomics and Molecular Medicine, School of Systems Biology, George Mason University Manassas, VA, USA.
authors.forename: Valerie
authors.initials: V
authors.lastname: Calvert
authors.affiliation: Center for Applied Proteomics and Molecular Medicine, School of Systems Biology, George Mason University Manassas, VA, USA.
authors.forename: Emanuel F
authors.initials: EF
authors.lastname: Petricoin
authors.affiliation: National Center for Biodefense and Infectious Diseases and School of Systems Biology, George Mason University Manassas, VA, USA.
authors.forename: Liana
authors.initials: L
authors.lastname: Kramer
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases Frederick, MD, USA.
authors.forename: Martha L
authors.initials: ML
authors.lastname: Hale
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases Frederick, MD, USA.
authors.forename: Sina
authors.initials: S
authors.lastname: Bavari
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases Frederick, MD, USA.
authors.forename: Rekha G
authors.initials: RG
authors.lastname: Panchal
authors.affiliation: National Center for Biodefense and Infectious Diseases and School of Systems Biology, George Mason University Manassas, VA, USA.
authors.forename: Ramin M
authors.initials: RM
authors.lastname: Hakami
keywords: RPMA; Yersinia pestis; apoptosis and autophagy; cell growth; host response; phosphorylation changes; proteomics; signaling pathways