The SecA2 auxiliary protein secretion system is required for secretion of virulence promoting proteins from a number of Gram-positive pathogens, including Bacillus anthracis (category A) and Listeria monocytogenes (category B). We identified two SecA2-dependent autolytic proteins that promote virulence of L. monocytogenes in infected animals, yet do not affect the growth of this bacterium in tissue culture cells. The virulence of engineered bacterial mutants lacking p60 was restored by expression of full-length p60, but not by expression of a truncated, catalytically inactive protein. The catalytic specificity of p60 predicts that it digests peptidoglycan (PGN) to generate or destroy immune modulating PGN fragments (muropeptides), including respectively muramyl di- and tri-peptides (MDP and MTP). MDP and MTP influence mammalian cell cytokine responses by acting on cytosolic proteins of the Nod family. We have found that p60- expressing bacteria and small molecules released from these bacteria enhance the induction of specific immune-regulatory cytokines by macrophages. In this grant proposal we investigate how p60 promotes virulence and affects host innate immune responses to infection.
Our first Aim will identify features of p60 that are required for PGN digestion and for its effects on bacterial virulence and cytokine gene expression.
Our second Aim i nvestigates the structure and phylogenetic distribution of a p60-dependent biologically active muropeptide or small molecule and tests whether responses to this molecule require known muropeptide-responsive Nod family proteins. For our third Aim, we investigate a potential mechanism for p60's effects on bacterial virulence by determining how expression of p60 and cytokines induced by p60 affect macrophage responses to activating stimuli. Our studies will define the mechanisms by which this bacterial autolysin contributes to the virulence of a clinically important bacterial pathogen and begin to explore whether similar mechanisms promote virulence of other Gram-positive pathogens, including potential agents of bioterrorism. The mechanisms used by pathogenic bacteria to cause disease include strategies to subvert host immune responses. A subversive strategy that may be common to a number of deadly bacteria is studied in this grant. Our studies will define the molecular basis for this strategy of immune subversion and may thus reveal novel therapeutic avenues to modulate inflammation during bacterial infection, vaccination, and chronic inflammatory diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI065638-05
Application #
7795786
Study Section
Host Interactions with Bacterial Pathogens Study Section (HIBP)
Program Officer
Mills, Melody
Project Start
2006-04-01
Project End
2011-07-31
Budget Start
2010-04-01
Budget End
2011-07-31
Support Year
5
Fiscal Year
2010
Total Cost
$367,781
Indirect Cost
Name
National Jewish Health
Department
Type
DUNS #
076443019
City
Denver
State
CO
Country
United States
Zip Code
80206
Mills, Charles D; Lenz, Laurel L; Harris, Robert A (2016) A Breakthrough: Macrophage-Directed Cancer Immunotherapy. Cancer Res 76:513-6
Mills, Charles D; Lenz, Laurel L; Ley, Klaus (2015) Macrophages at the fork in the road to health or disease. Front Immunol 6:59
Kedl, Ross M; Wysocki, Lawrence J; Janssen, William J et al. (2014) General parity between trio and pairwise breeding of laboratory mice in static caging. J Immunol 193:4757-60
Mills, Charles D; Thomas, Anita C; Lenz, Laurel L et al. (2014) Macrophage: SHIP of Immunity. Front Immunol 5:620
Czaja, Christopher A; Merkel, Patricia A; Chan, Edward D et al. (2014) Rituximab as successful adjunct treatment in a patient with disseminated nontuberculous mycobacterial infection due to acquired anti-interferon-? autoantibody. Clin Infect Dis 58:e115-8
Kearney, Staci J; Delgado, Christine; Eshleman, Emily M et al. (2013) Type I IFNs downregulate myeloid cell IFN-? receptor by inducing recruitment of an early growth response 3/NGFI-A binding protein 1 complex that silences ifngr1 transcription. J Immunol 191:3384-92
Kearney, Staci; Delgado, Christine; Lenz, Laurel L (2013) Differential effects of type I and II interferons on myeloid cells and resistance to intracellular bacterial infections. Immunol Res 55:187-200
Williams, Matthew A; Schmidt, Rebecca L; Lenz, Laurel L (2012) Early events regulating immunity and pathogenesis during Listeria monocytogenes infection. Trends Immunol 33:488-95
Schmidt, Rebecca L; Lenz, Laurel L (2012) Distinct licensing of IL-18 and IL-1? secretion in response to NLRP3 inflammasome activation. PLoS One 7:e45186
Cole, Caroline; Thomas, Stacey; Filak, Holly et al. (2012) Nitric oxide increases susceptibility of Toll-like receptor-activated macrophages to spreading Listeria monocytogenes. Immunity 36:807-20

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