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Haemophilus haemolyticus and nontypeableHaemophilus influenzae (NTHi) are closely related upper airway commensal bacteria that are difficult to distinguish phenotypically. NTHi causes upper and lower airway tract infections in individuals with compromised airways, whileH. haemolyticus rarely causes such infections. The lipooligosaccharide (LOS) is an outer membrane component of both species and plays a role in NTHi pathogenesis. In this study, comparative analyses of the LOS structures and corresponding biosynthesis genes were performed. Mass spectrometric and immunochemical analyses showed that NTHi LOS contained terminal sialic acid more frequently and to a higher extent thanH. haemolyticus LOS did. Genomic analyses of 10 strains demonstrated thatH. haemolyticus lacked the sialyltransferase geneslic3A andlic3B (9/10) andsiaA (10/10), but all strains contained the sialic acid uptake genessiaP andsiaT (10/10). However, isothermal titration calorimetry analyses of SiaP from twoH. haemolyticus strains showed a 3.4- to 7.3-fold lower affinity for sialic acid compared to that of NTHi SiaP. Additionally, mass spectrometric and immunochemical analyses showed that the LOS fromH. haemolyticus contained phosphorylcholine (ChoP) less frequently than the LOS from NTHi strains. These differences observed in the levels of sialic acid and ChoP incorporation in the LOS structures fromH. haemolyticus and NTHi may explain some of the differences in their propensities to cause disease.

infection and immunity

Comparative Analyses of the Lipooligosaccharides from Nontypeable Haemophilus influenzae and Haemophilus haemolyticus Show Differences in Sialic Acid and Phosphorylcholine Modifications