Nontypeable Haemophilus influenzae (NTHi) commonly colonize the mucosal surfaces of the human upper respiratory tract. For the most part, this species resides in that environment in a benign (symbiotic?) relationship with the human host. With pathophysiologic changes (obstruction of outflow ducts, loss of the mucociliary elevator, viral infection, etc.) in the airway anatomy and mucosal surface, NTHi can cause human upper and lower tract respiratory infections. The serum resistant phenotype appears to be a prerequisite for NTHi to initiate systemic infection (acute bronchitis, otitis media, etc). Studies from our laboratory and others have shown that sialic acid is important in the biology of many NTHi strains. Sialylation of LOS increases resistance to the bactericidal effect of normal human serum and is an important constituent of the NTHi biofilm. NTHi cannot synthesize sialic acid and must acquire it from its environment. Free sialic acid is quite limited in the human respiratory tract while sialoglycoproteins, such as mucin, are abundant. In our progress report, we have demonstrated that NTHi can remove sialic acid from mucin and incorporate this sugar into its LOS. The mechanism by which this is accomplished is unknown. Analysis of the 13 annotated NTHi genomes indicate that there are no genes homologous to any known eukaryotic or prokaryotic neuraminidase. We have also found that about 40% of NTHi strains do not incorporate sialic acid into their LOS. The factors determining NTHi LOS-based serum resistance and sensitivity are poorly defined. Better definition may lead to novel vaccines approaches. We have recently performed an NTHi human colonization experiment with NTHi 2019strR. The NTHi 2019strR strains recovered from 4 patients six days after infection demonstrated genomic changes after 454 pyrosequencing of these strains and NTHI 2019strR. The hypotheses underlying the studies in this proposal are: A) NTHi can scavenge sialic acid from human mucin by a unique glycosidase system, B) the LOS is a major determinant of serum resistance in NTHi including strains which do not sialylate their LOS, C) NTHi residence in the human nasopharynx selects for specific LOS phase variants and D) the transformability of NTHi allows introduction of foreign DNA into the NTHi genome. The following specific aims are proposed to resolve these hypotheses: 1) Identification of the mechanism by which sialic acid is removed from human mucin, 2) Comparative analysis of the LOS of selected serum resistant NTHi and serum sensitive NTHi strains which cannot sialylate their LOS to determine if specific LOS glycoforms are the basis of serum sensitivity and 3) Analysis of the phase-variation state of genes involved in LPS biosynthesis and the uptake and expression of foreign DNA in NTHi strain 2019strR recovered from patients during human colonization experiments.
This proposal describes studies which will elucidate the mechanism(s) by which nontypeable Haemophilus influenzae (NTHi) scavenges sialic acid from airway glycoproteins. Studies are also proposed which will elucidate the role of the non-sialylated lipooligosaccaride in resistance to serum bactericidal activity. Finally, studies of isolates from human subjects colonized with NTHi 2019strR will examine the role of phase variation on LOS structure and determine if foreign DNA is acquired by NTHi during colonization.
|Post, Deborah M B; Yu, Liping; Krasity, Benjamin C et al. (2012) O-antigen and core carbohydrate of Vibrio fischeri lipopolysaccharide: composition and analysis of their role in Euprymna scolopes light organ colonization. J Biol Chem 287:8515-30|
|Sandal, Indra; Inzana, Thomas J; Molinaro, Antonio et al. (2011) Identification, structure, and characterization of an exopolysaccharide produced by Histophilus somni during biofilm formation. BMC Microbiol 11:186|
|Phillips, Nancy J; Adin, Dawn M; Stabb, Eric V et al. (2011) The lipid A from Vibrio fischeri lipopolysaccharide: a unique structure bearing a phosphoglycerol moiety. J Biol Chem 286:21203-19|
|Johnston, Jason W; Shamsulddin, Haider; Miller, Anne-Frances et al. (2010) Sialic acid transport and catabolism are cooperatively regulated by SiaR and CRP in nontypeable Haemophilus influenzae. BMC Microbiol 10:240|
|Johansen, Eric B; Szoka, Francis C; Zaleski, Anthony et al. (2010) Utilizing the O-antigen lipopolysaccharide biosynthesis pathway in Escherichia coli to interrogate the substrate specificities of exogenous glycosyltransferase genes in a combinatorial approach. Glycobiology 20:763-74|
|Sandal, Indra; Shao, Jian Q; Annadata, Satish et al. (2009) Histophilus somni biofilm formation in cardiopulmonary tissue of the bovine host following respiratory challenge. Microbes Infect 11:254-63|
|Johnston, Jason W; Coussens, Nathan P; Allen, Simon et al. (2008) Characterization of the N-acetyl-5-neuraminic acid-binding site of the extracytoplasmic solute receptor (SiaP) of nontypeable Haemophilus influenzae strain 2019. J Biol Chem 283:855-65|
|Johnston, Jason W; Zaleski, Anthony; Allen, Simon et al. (2007) Regulation of sialic acid transport and catabolism in Haemophilus influenzae. Mol Microbiol 66:26-39|
|Starner, Timothy D; Zhang, Niu; Kim, Gunhee et al. (2006) Haemophilus influenzae forms biofilms on airway epithelia: implications in cystic fibrosis. Am J Respir Crit Care Med 174:213-20|
|Erwin, Alice L; Allen, Simon; Ho, Derek K et al. (2006) Role of lgtC in resistance of nontypeable Haemophilus influenzae strain R2866 to human serum. Infect Immun 74:6226-35|
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