Current vaccines against Haemophilus influenzae type b have reduced the incidence of H. influenzae type b invasive diseases. However, they provide no protection against nontypable strains which remain a common cause of otitis media, the most common infection of childhood, and pneumonia. H. influenzae has an absolute growth requirement for heme and the human body is its sole niche. We have characterized several of the heme and iron acquisition systems of H. influenzae and shown that their expression is regulated by iron/heme levels in the growth media. In addition we have shown that the genes are expressed in vivo in both humans and animal models, indicating that the in vivo environment is similarly heme restricted. However our understanding of the systems involved in the acquisition of this essential nutrient and their regulation remains incomplete. We propose a genomics analysis to define the genes whose expression is regulated by iron and heme levels (the FeHm regulon) and to determine their role in virulence in animal models of disease. The FeHm regulon has been defined in one sequenced H. influenzae strain using microarray technology, and we will use the same approach to define the FeHm regulon in two additional sequenced strains. Following identification of the genes we will construct pools of signature-tagged mutants for analysis in vivo in three animal models (the chinchilla model of otitis media and nasopharyngeal colonization, the rat model of invasive disease and the rat model of persistence in the lung). Mutants will additionally be compared to wildtype strains for in vitro growth with various heme sources, as well as protection against heme toxicity and oxidative damage and for their ability to form biofilms. These studies will provide a framework for advancing our understanding of the iron/heme acquisition systems of H. influenzae with further potential for providing preventive and treatment strategies for H. influenzae disease. PROJECT NARRATIVE Haemophilus influenzae is a major cause of childhood ear infections and pneumonia in adults who have other conditions, such as cystic fibrosis or HIV/AIDS, predisposing them to infections. We are seeking to understand what makes Haemophilus influenzae able to cause these infections and especially how it gets important nutrients, such as iron, when it is causing an infection. Understanding these mechanisms has the potential to lead to novel treatments for and/or vaccines to prevent Haemophilus influenzae infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI029611-20
Application #
8260862
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Taylor, Christopher E,
Project Start
1990-12-01
Project End
2014-04-30
Budget Start
2012-05-01
Budget End
2014-04-30
Support Year
20
Fiscal Year
2012
Total Cost
$323,066
Indirect Cost
$102,543
Name
University of Oklahoma Health Sciences Center
Department
Pediatrics
Type
Schools of Medicine
DUNS #
878648294
City
Oklahoma City
State
OK
Country
United States
Zip Code
73117
Morton, Daniel J; Smith, Ann; VanWagoner, Timothy M et al. (2007) Lipoprotein e (P4) of Haemophilus influenzae: role in heme utilization and pathogenesis. Microbes Infect 9:932-9
Morton, Daniel J; Seale, Thomas W; Madore, Larissa L et al. (2007) The haem-haemopexin utilization gene cluster (hxuCBA) as a virulence factor of Haemophilus influenzae. Microbiology 153:215-24
Morton, Daniel J; VanWagoner, Timothy M; Seale, Thomas W et al. (2006) Differential utilization by Haemophilus influenzae of haemoglobin complexed to the three human haptoglobin phenotypes. FEMS Immunol Med Microbiol 46:426-32
Morton, Daniel J; Van Wagoner, Timothy M; Seale, Thomas W et al. (2006) Utilization of myoglobin as a heme source by Haemophilus influenzae requires binding of myoglobin to haptoglobin. FEMS Microbiol Lett 258:235-40
Whitby, Paul W; Vanwagoner, Timothy M; Seale, Thomas W et al. (2006) Transcriptional profile of Haemophilus influenzae: effects of iron and heme. J Bacteriol 188:5640-5
Seale, Thomas W; Morton, Daniel J; Whitby, Paul W et al. (2006) Complex role of hemoglobin and hemoglobin-haptoglobin binding proteins in Haemophilus influenzae virulence in the infant rat model of invasive infection. Infect Immun 74:6213-25
Morton, Daniel J; Madore, Larissa L; Smith, Ann et al. (2005) The heme-binding lipoprotein (HbpA) of Haemophilus influenzae: role in heme utilization. FEMS Microbiol Lett 253:193-9
Morton, Daniel J; Smith, Ann; Ren, Zhen et al. (2004) Identification of a haem-utilization protein (Hup) in Haemophilus influenzae. Microbiology 150:3923-33
Morton, Daniel J; Bakaletz, Lauren O; Jurcisek, Joseph A et al. (2004) Reduced severity of middle ear infection caused by nontypeable Haemophilus influenzae lacking the hemoglobin/hemoglobin-haptoglobin binding proteins (Hgp) in a chinchilla model of otitis media. Microb Pathog 36:25-33
VanWagoner, Timothy M; Whitby, Paul W; Morton, Daniel J et al. (2004) Characterization of three new competence-regulated operons in Haemophilus influenzae. J Bacteriol 186:6409-21

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