The innate immune system that defends mammals from many Gram-negative bacteria is triggered when host cells sense the lipid A moiety of the bacterial cell wall lipopolysaccharide (LPS). The resulting inflammatory response then usually kills the bacteria. LPS-induced inflammation can also be harmful, however, and animals have numerous ways to limit its magnitude and duration. One important control mechanism is a highly conserved lipase, acyloxyacyl hydrolase (AOAH) that selectively removes from LPS the fatty acyl chains that are required for sensing by MD-2--TLR4, the mammalian LPS receptor. Mice that have a disabling mutation in the AOAH gene respond to low doses of LPS by producing large quantities of antibodies and experiencing prolonged immunosuppression. If the LPS are given intravenously, they also develop striking enlargement of the liver. LPS that is not deacylated by AOAH remains stimulatory for weeks in vivo. We now request continuing support to test the hypothesis that providing AOAH will prevent prolonged reactions to LPS and Gram-negative bacteria in vivo, thereby benefiting animals with serious Gram-negative bacterial infections.
In Specific Aim 1, we will find out how LPS deacylation by phagocytes in a subcutaneous tissue bed regulates its ability to traffic to lymph nodes and interact with a second LPS-responsive target cell, the B lymphocyte. These studies should provide the first quantitative information about how LPS processing by phagocytes prevents activation of other cells in extravascular tissue spaces.
In Specific Aim 2 we will define AOAH's role in the liver, the organ that normally detoxifies most bloodborne endotoxin, and determine the cellular basis for the persistent hepatomegaly that LPS induces in Aoah-/- mice. These experiments should indicate how AOAH prevents prolonged LPS- induced reactions in the intravascular (systemic) compartment.
In Specific Aim 3, we will use three complementary strategies to find out whether increasing LPS deacylation in vivo can protect animals with serious Gram-negative bacterial infections from dying and/or prevent prolonged post-infection immunosuppression. In addition to defining the enzyme's potential for controlling responses to LPS in vivo, the proposed studies should break new ground in understanding the """"""""recovery"""""""" phase of Gram- negative bacterial infections, an important yet understudied aspect of bacterial pathogenesis and host defense.

Public Health Relevance

- relevance for public health: Gram-negative bacteria cause a large fraction of the serious infections that occur in the U.S. today. In patients who are infected with these bacteria, a bacterial molecule called endotoxin is the most important trigger for the lethal condition known as septic shock. Over 100,000 Americans die each year from septic shock due to these bacteria. The subject of our research is a human enzyme that can detoxify bacterial endotoxin. Having discovered the enzyme, acyloxyacyl hydrolase (AOAH), and studied it intensively, we want now to find out if it can prevent or treat this lethal reaction to Gram-negative bacterial infection. Since the currently available drugs for septic shock are not very effective, finding that AOAH can prevent or reverse the septic reaction could have an enormous impact on public health. No other laboratory has worked as intensively on AOAH or has the ability to do these tests.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI018188-29
Application #
7753844
Study Section
Host Interactions with Bacterial Pathogens Study Section (HIBP)
Program Officer
Korpela, Jukka K
Project Start
1981-08-01
Project End
2011-12-31
Budget Start
2010-01-01
Budget End
2010-12-31
Support Year
29
Fiscal Year
2010
Total Cost
$468,011
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
Lu, Mingfang; Varley, Alan W; Munford, Robert S (2013) Persistently active microbial molecules prolong innate immune tolerance in vivo. PLoS Pathog 9:e1003339
Shao, Baomei; Munford, Robert S; Kitchens, Richard et al. (2012) Hepatic uptake and deacylation of the LPS in bloodborne LPS-lipoprotein complexes. Innate Immun 18:825-33
Shao, Baomei; Kitchens, Richard L; Munford, Robert S et al. (2011) Prolonged hepatomegaly in mice that cannot inactivate bacterial endotoxin. Hepatology 54:1051-62
Lu, Mingfang; Munford, Robert S (2011) The transport and inactivation kinetics of bacterial lipopolysaccharide influence its immunological potency in vivo. J Immunol 187:3314-20
Thompson, Patricia A; Gauthier, Karine C; Varley, Alan W et al. (2010) ABCA1 promotes the efflux of bacterial LPS from macrophages and accelerates recovery from LPS-induced tolerance. J Lipid Res 51:2672-85
Ojogun, Noredia; Kuang, Tang-Yong; Shao, Baomei et al. (2009) Overproduction of acyloxyacyl hydrolase by macrophages and dendritic cells prevents prolonged reactions to bacterial lipopolysaccharide in vivo. J Infect Dis 200:1685-93
Munford, Robert; Lu, Mingfang; Varley, Alan (2009) Chapter 2: Kill the bacteria...and also their messengers? Adv Immunol 103:29-48
Lu, Mingfang; Varley, Alan W; Ohta, Shoichiro et al. (2008) Host inactivation of bacterial lipopolysaccharide prevents prolonged tolerance following gram-negative bacterial infection. Cell Host Microbe 4:293-302
Gioannini, Theresa L; Teghanemt, Athmane; Zhang, DeSheng et al. (2007) Endotoxin-binding proteins modulate the susceptibility of bacterial endotoxin to deacylation by acyloxyacyl hydrolase. J Biol Chem 282:7877-84
Shao, Baomei; Lu, Mingfang; Katz, Steven C et al. (2007) A host lipase detoxifies bacterial lipopolysaccharides in the liver and spleen. J Biol Chem 282:13726-35

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