Outer membrane protein p5 is required for resistance of nontypeable haemophilus influenzae to both the classical and alternative complement pathways

Infection and Immunity

Volumn 82, Issue 2, 2014, Pages 640-649

  Rosadini, Charles V ^a   Ram, Sanjay ^a   Akerleya, Brian J ^a,b  

^a UNIVERSITY OF MASSACHUSETTS MEDICAL SCHOOL   (United States)

^b UNIVERSITY OF MISSISSIPPI SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

COMPLEMENT COMPONENT C3; COMPLEMENT COMPONENT C3B; COMPLEMENT COMPONENT C4; COMPLEMENT COMPONENT C4B; COMPLEMENT FACTOR H; IMMUNOGLOBULIN M; OUTER MEMBRANE PROTEIN; OUTER MEMBRANE PROTEIN P5; UNCLASSIFIED DRUG;

ARTICLE; BACTERIAL GROWTH; BACTERIAL IMMUNITY; BACTERIAL MUTATION; BACTERIAL STRAIN; BACTERIAL VIRULENCE; BACTERIUM ISOLATE; BACTERIUM MUTANT; CELL SURFACE; COMPLEMENT ACTIVATION; COMPLEMENT ALTERNATIVE PATHWAY; COMPLEMENT CLASSICAL PATHWAY; CONTROLLED STUDY; GENOTYPE; HAEMOPHILUS INFLUENZAE; HOST PATHOGEN INTERACTION; HUMAN; NONHUMAN; PHENOTYPE; PRIORITY JOURNAL; PROTEIN EXPRESSION; PROTEIN FUNCTION; SERUM; SERUM BACTERICIDAL ACTIVITY;

ADOLESCENT; ADULT; AGED; ANTIBODIES, BACTERIAL; BACTERIAL OUTER MEMBRANE PROTEINS; BLOOD BACTERICIDAL ACTIVITY; COMPLEMENT PATHWAY, ALTERNATIVE; COMPLEMENT PATHWAY, CLASSICAL; COMPLEMENT SYSTEM PROTEINS; HAEMOPHILUS INFLUENZAE; HUMANS; IMMUNOGLOBULIN M; MICROBIAL VIABILITY; MIDDLE AGED; PROTEIN BINDING; YOUNG ADULT;

EID: 84892977856     PISSN: 00199567     EISSN: 10985522     Source Type: Journal    
DOI: 10.1128/IAI.01224-13     Document Type: Article

References (77)

1. CDC. 2002. Progress toward elimination of Haemophilus influenzae type b invasive disease among infants and children-United States, 1998-2000. JAMA 287:2206-2207. http://dx.doi.org/10.1001/jama.1994 .03510400017006.
2. Poolman J, Bakaletz L, Cripps A, Denoel P, Forsgren A, Kyd J. 2000. Developing a nontypeable Haemophilus influenzae (NTHi) vaccine. Vaccine 19:S108-S115. http://dx.doi.org/10.1016/S0264-410X(00)00288-7.
3. Murphy TF, Faden H, Bakaletz LO, Kyd JM, Forsgren A, Campos J, Virji M, Pelton SI. 2009. Nontypeable Haemophilus influenzae as a pathogen in children. Pediatr. Infect. Dis. J. 28:43-48. http://dx.doi.org/10.1097 /INF.0b013e318184dba2.
4. Murphy TF. 2003. Respiratory infections caused by non-typeable Haemophilus influenzae. Curr. Opin. Infect. Dis. 16:129-134. http://dx.doi .org/10.1097/00001432-200304000-00009.
5. Mannino DM, Kiriz VA. 2006. Changing the burden of COPD mortality. Int. J. Chron. Obstruct. Pulmon. Dis. 1:219-233. http://www.ncbi.nlm .nih.gov/pmc/articles/PMC2707151/.
6. Murphy TF, Brauer AL, Schiffmacher AT, Sethi S. 2004. Persistent colonization by Haemophilus influenzae in chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 170:266-272. http://dx.doi.org/10 .1164/rccm.200403-354OC.
7. Murphy TF, Sethi S. 2002. Chronic obstructive pulmonary disease: role of bacteriaandguide to antibacterial selection in the older patient. Drugs Aging 19:761- 775. http://dx.doi.org/10.2165/00002512-200219100-00005.
8. Sethi S, Murphy TF. 2001. Bacterial infection in chronic obstructive pulmonary disease in 2000: a state-of-the-art review. Clin. Microbiol. Rev. 14:336-363. http://dx.doi.org/10.1128/CMR.14.2.336-363.2001.
9. Saiman L. 2004. Microbiology of early CF lung disease. Paediatr. Respir. Rev. 5(Suppl A):S367-S369. http://dx.doi.org/10.1016/S1526-0542(04) 90065-6.
10. Moller LV, Regelink AG, Grasselier H, Dankert-Roelse JE, Dankert J, van Alphen L. 1995. Multiple Haemophilus influenzae strains and strain variants coexist in the respiratory tract of patients with cystic fibrosis. J. Infect. Dis. 172:1388-1392. http://dx.doi.org/10.1093/infdis/172.5.1388.
11. Gilligan PH. 1991. Microbiology of airway disease in patients with cystic fibrosis. Clin. Microbiol. Rev. 4:35-51.
12. O'Neill JM, St Geme JW, III, Cutter D, Adderson EE, Anyanwu J, Jacobs RF, Schutze GE. 2003. Invasive disease due to nontypeable Haemophilus influenzae among children in Arkansas. J. Clin. Microbiol. 41: 3064-3069. http://dx.doi.org/10.1128/JCM.41.7.3064-3069.2003.
13. Ito T, Shibata H, Nakazawa M, Myokai M, Ikegaya K, Tsuchiya K, Kamimaki T. 2011. Meningitis and septicemia caused by nontypeable Haemophilus influenzae in a previously healthy 2-year-old girl. J. Infect. Chemother. 17:559-562. http://dx.doi.org/10.1007/s10156-011-0213-6.
14. Cuthill SL, Farley MM, Donowitz LG. 1999. Nontypable Haemophilus influenzae meningitis. Pediatr. Infect. Dis. J. 18:660-662. http://dx.doi .org/10.1097/00006454-199907000-00024.
15. Nizet V, Colina KF, Almquist JR, Rubens CE, Smith AL. 1996. A virulent nonencapsulated Haemophilus influenzae. J. Infect. Dis. 173:180- 186. http://dx.doi.org/10.1093/infdis/173.1.180.
16. Campos J, Hernando M, Roman F, Perez-Vazquez M, Aracil B, Oteo J, Lazaro E, de Abajo F. 2004. Analysis of invasive Haemophilus influenzae infections after extensive vaccination against H. influenzae type b. J. Clin. Microbiol. 42:524-529. http://dx.doi.org/10.1128/JCM.42.2.524-529.2004.
17. Cerquetti M, Ciofi degli Atti ML, Renna G, Tozzi AE, Garlaschi ML, Mastrantonio P. 2000. Characterization of non-type B Haemophilus influenzae strains isolated from patients with invasive disease. The HI Study Group. J. Clin. Microbiol. 38:4649-4652. http://jcm.asm.org/content/38 /12/4649.long.
18. Walport MJ. 2001. Complement. First of two parts. N. Engl. J. Med. 344:1058-1066. http://dx.doi.org/10.1056/NEJM200104053441406.
19. Narkio-Makela M, Hellwage J, Tahkokallio O, Meri S. 2001. Complement- regulator factorHand related proteins in otitis media with effusion. Clin. Immunol. 100:118-126. http://dx.doi.org/10.1006/clim.2001.5043.
20. Narkio-Makela M, Teppo AM, Meri S. 2000. Complement C3 cleavage and cytokines interleukin-1beta and tumor necrosis factor-alpha in otitis media with effusion. Laryngoscope 110:1745-1749. http://dx.doi.org/10 .1097/00005537-200010000-00035.
21. Van Zele T, Coppieters F, Gevaert P, Holtappels G, Van Cauwenberge P, Bachert C. 2009. Local complement activation in nasal polyposis. Laryngoscope 119:1753-1758. http://dx.doi.org/10.1002/lary.20484.
22. Andersson M, Michel L, Llull JB, Pipkorn U. 1994. Complement activation on the nasal mucosal surface-a feature of the immediate allergic reaction in the nose. Allergy 49:242-245. http://dx.doi.org/10.1111/j.1398 -9995.1994.tb02656.x.
23. Marc MM, Korosec P, Kosnik M, Kern I, Flezar M, Suskovic S, Sorli J. 2004. Complement factors c3a, c4a, and c5a in chronic obstructive pulmonary disease and asthma. Am. J. Respir. Cell Mol. Biol. 31:216-219. http://dx.doi.org/10.1165/rcmb.2003-0394OC.
24. Hallstrom T, Resman F, Ristovski M, Riesbeck K. 2010. Binding of complement regulators to invasive nontypeable Haemophilus influenzae isolates is not increased compared to nasopharyngeal isolates, but serum resistance is linked to disease severity. J. Clin. Microbiol. 48:921-927. http: //dx.doi.org/10.1128/JCM.01654-09.
25. Du Clos TW. 2000. Function of C-reactive protein. Ann. Med. 32:274- 278. http://dx.doi.org/10.3109/07853890009011772.
26. Pangburn MK, Muller-Eberhard HJ. 1983. Initiation of the alternative complement pathway due to spontaneous hydrolysis of the thioester of C3. Ann. N. Y. Acad. Sci. 421:291-298. http://dx.doi.org/10.1111/j.1749 -6632.1983.tb18116.x.
27. Wong SM, St Michael F, Cox A, Ram S, Akerley BJ. 2011. ArcAregulated glycosyltransferase Lic2B promotes complement evasion and pathogenesis of nontypeable Haemophilus influenzae. Infect. Immun. 79: 1971-1983. http://dx.doi.org/10.1128/IAI.01269-10.
28. Ho DK, Ram S, Nelson KL, Bonthuis PJ, Smith AL. 2007. lgtC expression modulates resistance to C4b deposition on an invasive nontypeable Haemophilus influenzae. J. Immunol. 178:1002-1012. http://www .jimmunol.org/content/178/2/1002.long.
29. Hood DW, Makepeace K, Deadman ME, Rest RF, Thibault P, Martin A, Richards JC, Moxon ER. 1999. Sialic acid in the lipopolysaccharide of Haemophilus influenzae: strain distribution, influence on serum resistance and structural characterization. Mol. Microbiol. 33:679-692. http://dx .doi.org/10.1046/j.1365-2958.1999.01509.x.
30. Figueira MA, Ram S, Goldstein R, Hood DW, Moxon ER, Pelton SI. 2007. Role of complement in defense of the middle ear revealed by restoring the virulence of nontypeable Haemophilus influenzae siaB mutants. Infect. Immun. 75:325-333. http://dx.doi.org/10.1128/IAI.01054-06.
31. Hallstrom T, Riesbeck K. 2010. Haemophilus influenzae and the complement system. Trends Microbiol. 18:258-265. http://dx.doi.org/10.1016/j .tim.2010.03.007.
32. Su YC, Jalalvand F, Morgelin M, Blom AM, Singh B, Riesbeck K. 2013. Haemophilus influenzae acquires vitronectin via the ubiquitous protein F to subvert host innate immunity. Mol. Microbiol. 87:1245-1266. http://dx .doi.org/10.1111/mmi.12164.
33. Bardwell JC, McGovern K, Beckwith J. 1991. Identification of a protein required for disulfide bond formation in vivo. Cell 67:581-589. http://dx .doi.org/10.1016/0092-8674(91)90532-4.
34. Rosadini CV, Wong SM, Akerley BJ. 2008. The periplasmic disulfide oxidoreductase DsbA contributes to Haemophilus influenzae pathogenesis. Infect. Immun. 76:1498-1508. http://dx.doi.org/10.1128/IAI.01378-07.
35. Rosadini CV, Gawronski JD, Raimunda D, Arguello JM, Akerley BJ. 2011. A novel zinc binding system, ZevAB, is critical for survival of nontypeable Haemophilus influenzae in a murine lung infection model. Infect. Immun. 79:3366-3376. http://dx.doi.org/10.1128/IAI.05135-11.
36. Munson RS, Jr, Grass S, West R. 1993. Molecular cloning and sequence of the gene for outer membrane protein P5 of Haemophilus influenzae. Infect. Immun. 61:4017-4020.
37. Weiser JN, Gotschlich EC. 1991. Outer membrane protein A (OmpA) contributes to serum resistance and pathogenicity of Escherichia coli K-1. Infect. Immun. 59:2252-2258.
38. Prasadarao NV, Blom AM, Villoutreix BO, Linsangan LC. 2002. A novel interaction of outer membrane protein A with C4b binding protein mediates serum resistance of Escherichia coli K1. J. Immunol. 169:6352-6360. http://www.jimmunol.org/content/169/11/6352.long.
39. Wooster DG, Maruvada R, Blom AM, Prasadarao NV. 2006. Logarith- mic phase Escherichia coli K1 efficiently avoids serum killing by promoting C4bp-mediated C3b and C4b degradation. Immunology 117:482-493. http://dx.doi.org/10.1111/j.1365-2567.2006.02323.x.
40. Webb DC, Cripps A. 1998. Secondary structure and molecular analysis of interstrain variability in the P5 outer-membrane protein of non-typable Haemophilus influenzae isolated from diverse anatomical sites. J. Med. Microbiol. 47:1059-1067. http://dx.doi.org/10.1099/00222615-47 -12-1059.
41. Sirakova T, Kolattukudy PE, Murwin D, Billy J, Leake E, Lim D, DeMaria T, Bakaletz L. 1994. Role of fimbriae expressed by nontypeable Haemophilus influenzae in pathogenesis of and protection against otitis media and relatedness of the fimbrin subunit to outer membrane protein A. Infect. Immun. 62:2002-2020.
42. Reddy MS, Bernstein JM, Murphy TF, Faden HS. 1996. Binding between outer membrane proteins of nontypeable Haemophilus influenzae and human nasopharyngeal mucin. Infect. Immun. 64:1477-1479.
43. Jiang Z, Nagata N, Molina E, Bakaletz LO, Hawkins H, Patel JA. 1999. Fimbria-mediated enhanced attachment of nontypeable Haemophilus influenzae to respiratory syncytial virus-infected respiratory epithelial cells. Infect. Immun. 67:187-192.
44. Miyamoto N, Bakaletz LO. 1996. Selective adherence of non-typeable Haemophilus influenzae (NTHi) to mucus or epithelial cells in the chinchilla eustachian tube and middle ear. Microb. Pathog. 21:343-356. http: //dx.doi.org/10.1006/mpat.1996.0067.
45. Hill DJ, Toleman MA, Evans DJ, Villullas S, Van Alphen L, Virji M. 2001. The variable P5 proteins of typeable and non-typeable Haemophilus influenzae target human CEACAM1. Mol. Microbiol. 39:850-862. http: //dx.doi.org/10.1046/j.1365-2958.2001.02233.x.
46. Bookwalter JE, Jurcisek JA, Gray-Owen SD, Fernandez S, McGillivary G, Bakaletz LO. 2008. A carcinoembryonic antigen-related cell adhesion molecule 1 homologue plays a pivotal role in nontypeable Haemophilus influenzae colonization of the chinchilla nasopharynx via the outer membrane protein P5-homologous adhesin. Infect. Immun. 76:48-55. http: //dx.doi.org/10.1128/IAI.00980-07.
47. Wong SM, Akerley BJ. 2003. Inducible expression system and marker-linked mutagenesis approach for functional genomics of Haemophilus influenzae. Gene 316:177-186. http://dx.doi.org/10.1016/S0378-1119(03)00762-5.
48. Harrington JC, Wong SM, Rosadini CV, Garifulin O, Boyartchuk V, Akerley BJ. 2009. Resistance of Haemophilus influenzae to reactive nitrogen donors and gamma interferon-stimulated macrophages requires the formate-dependent nitrite reductase regulator-activated ytfE gene. Infect. Immun. 77:1945-1958. http://dx.doi.org/10.1128/IAI.01365-08.
49. Gawronski JD, Wong SM, Giannoukos G, Ward DV, Akerley BJ. 2009. Tracking insertion mutants within libraries by deep sequencing and a genome-wide screen for Haemophilus genes required in the lung. Proc. Natl. Acad. Sci. U. S. A. 106:16422-16427. http://dx.doi.org/10.1073/pnas .0906627106.
50. Barcak GJ, Chandler MS, Redfield RJ, Tomb JF. 1991. Genetic systems in Haemophilus influenzae. Methods Enzymol. 204:321-342.
51. Ausubel FM, Brent R, Kingston RE, Moore DE, Seidman JG, Smith JA, Struhl K (ed). 1995. Current protocols in molecular biology. John Wiley & Sons, Inc. New York, NY.
52. Ram S, Ngampasutadol J, Cox AD, Blom AM, Lewis LA, St Michael F, Stupak J, Gulati S, Rice PA. 2007. Heptose I glycan substitutions on Neisseria gonorrhoeae lipooligosaccharide influence C4b-binding protein binding and serum resistance. Infect. Immun. 75:4071-4081. http://dx .doi.org/10.1128/IAI.01109-06.
53. Ram S, Cox AD, Wright JC, Vogel U, Getzlaff S, Boden R, Li J, Plested JS, Meri S, Gulati S, Stein DC, Richards JC, Moxon ER, Rice PA. 2003. Neisserial lipooligosaccharide is a target for complement component C4b. Inner core phosphoethanolamine residues define C4b linkage specificity. J. Biol. Chem. 278:50853-50862. http://dx.doi.org/10.1074/jbc.M308364200.
54. Iida K, Mitomo K, Fujita T, Tamura N. 1987. Characterization of three monoclonal antibodies against C3 with selective specificities. Immunology 62:413-417.
55. Lewis LA, Ram S, Prasad A, Gulati S, Getzlaff S, Blom AM, Vogel U, Rice PA. 2008. Defining targets for complement components C4b and C3b on the pathogenic neisseriae. Infect. Immun. 76:339-350. http://dx .doi.org/10.1128/IAI.00613-07.
56. Ram S, Cullinane M, Blom AM, Gulati S, McQuillen DP, Monks BG, O'Connell C, Boden R, Elkins C, Pangburn MK. 2001. Binding of C4b-binding protein to porin. J. Exp. Med. 193:281-295. http://dx.doi.org /10.1084/jem.193.3.281.
57. Erwin AL, Bonthuis PJ, Geelhood JL, Nelson KL, McCrea KW, Gilsdorf JR, Smith AL. 2006. Heterogeneity in tandem octanucleotides within Haemophilus influenzae lipopolysaccharide biosynthetic gene losA affects serum resistance. Infect. Immun. 74:3408-3414. http://dx.doi.org/10 .1128/IAI.01540-05.
58. Griffin R, Bayliss CD, Herbert MA, Cox AD, Makepeace K, Richards JC, Hood DW, Moxon ER. 2005. Digalactoside expression in the lipopolysaccharide of Haemophilus influenzae and its role in intravascular survival. Infect. Immun. 73:7022-7026. http://dx.doi.org/10.1128/IAI.73.10 .7022-7026.2005.
59. Wong SM, Alugupalli KR, Ram S, Akerley BJ. 2007. The ArcA regulon and oxidative stress resistance in Haemophilus influenzae. Mol. Microbiol. 64:1375-1390. http://dx.doi.org/10.1111/j.1365-2958.2007.05747.x.
60. Priebe GP, Dean CR, Zaidi T, Meluleni GJ, Coleman FT, Coutinho YS, Noto MJ, Urban TA, Pier GB, Goldberg JB. 2004. The galU gene of Pseudomonas aeruginosa is required for corneal infection and efficient systemic spread following pneumonia but not for infection confined to the lung. Infect. Immun. 72:4224-4232. http://dx.doi.org/10.1128/IAI.72.7 .4224-4232.2004.
61. Williams BJ, Morlin G, Valentine N, Smith AL. 2001. Serum resistance in an invasive, nontypeable Haemophilus influenzae strain. Infect. Immun. 69:695-705. http://dx.doi.org/10.1128/IAI.69.2.695-705.2001.
62. Ram S, Lewis LA, Rice PA. 2010. Infections of people with complement deficiencies and patients who have undergone splenectomy. Clin. Microbiol. Rev. 23:740-780. http://dx.doi.org/10.1128/CMR.00048-09.
63. Winkelstein JA, Moxon ER. 1992. The role of complement in the host's defense against Haemophilus influenzae. J. Infect. Dis. 165(Suppl 1):S62- S65.
64. Nakamura S, Shchepetov M, Dalia AB, Clark SE, Murphy TF, Sethi S, Gilsdorf JR, Smith AL, Weiser JN. 2011. Molecular basis of increased serum resistance among pulmonary isolates of non-typeable Haemophilus influenzae. PLoS Pathog. 7:e1001247. http://dx.doi.org/10.1371/journal .ppat.1001247.
65. Langereis JD, Stol K, Schweda EK, Twelkmeyer B, Bootsma HJ, de Vries SP, Burghout P, Diavatopoulos DA, Hermans PW. 2012. Modified lipooligosaccharide structure protects nontypeable Haemophilus influenzae from IgM-mediated complement killing in experimental otitis media. mBio 3(4):e00079-12. http://dx.doi.org/10.1128/mBio.00079-12.
66. Berenson CS, Kruzel RL, Eberhardt E, Sethi S. 2013. Phagocytic dysfunction of human alveolar macrophages and severity of chronic obstructive pulmonary disease. J. Infect. Dis. 208:2036-2045. http://dx.doi.org /10.1093/infdis/jit400.
67. Janeway CA, Travers P, Walport M, Shlomchik MJ. 2001. Immunobiology: the immune system in health and disease, 5th ed. Garland Science, New York, NY.
68. Gnehm HE, Pelton SI, Gulati S, Rice PA. 1985. Characterization of antigens from nontypable Haemophilus influenzae recognized by human bactericidal antibodies. Role of Haemophilus outer membrane proteins. J. Clin. Invest. 75:1645-1658.
69. Abdullah M, Nepluev I, Afonina G, Ram S, Rice P, Cade W, Elkins C. 2005. Killing of dsrA mutants of Haemophilus ducreyi by normal human serum occurs via the classical complement pathway and is initiated by immunoglobulinMbinding. Infect. Immun. 73:3431-3439. http://dx.doi .org/10.1128/IAI.73.6.3431-3439.2005.
70. Hallstrom T, Zipfel PF, Blom AM, Lauer N, Forsgren A, Riesbeck K. 2008. Haemophilus influenzae interacts with the human complement inhibitor factor H. J. Immunol. 181:537-545. http://www.jimmunol.org /content/181/1/537.long.
71. Marti-Lliteras P, Lopez-Gomez A, Mauro S, Hood DW, Viadas C, Calatayud L, Morey P, Servin A, Linares J, Oliver A, Bengoechea JA, Garmendia J. 2011. Nontypable Haemophilus influenzae displays a prevalent surface structure molecular pattern in clinical isolates. PLoS One 6:e21133. http://dx.doi.org/10.1371/journal.pone.0021133.
72. Madico G, Welsch JA, Lewis LA, McNaughton A, Perlman DH, Costello CE, Ngampasutadol J, Vogel U, Granoff DM, Ram S. 2006. The meningococcal vaccine candidate GNA1870 binds the complement regulatory protein factor H and enhances serum resistance. J. Immunol. 177:501- 510. http://www.jimmunol.org/content/177/1/501.long.
73. Lewis LA, Ngampasutadol J, Wallace R, Reid JE, Vogel U, Ram S. 2010. The meningococcal vaccine candidate neisserial surface protein A (NspA) binds to factor H and enhances meningococcal resistance to complement. PLoS Pathog. 6:e1001027. http://dx.doi.org/10.1371/journal.ppat .1001027.
74. Zipfel PF, Hallstrom T, Hammerschmidt S, Skerka C. 2008. The complement fitness factor H: role in human diseases and for immune escape of pathogens, like pneumococci. Vaccine 26(Suppl 8):I67-I74. http://dx.doi .org/10.1016/j.vaccine.2008.11.015.
75. Kraiczy P, Stevenson B. 2013. Complement regulator-acquiring surface proteins of Borrelia burgdorferi: structure, function and regulation of gene expression. Ticks Tick Borne Dis. 4:26-34. http://dx.doi.org/10.1016/j .ttbdis.2012.10.039.
76. Luo S, Hoffmann R, Skerka C, Zipfel PF. 2013. Glycerol-3-phosphate dehydrogenase 2 is a novel factor H-, factor H-like protein 1-, and plasminogen- binding surface protein of Candida albicans. J. Infect. Dis. 207: 594-603. http://dx.doi.org/10.1093/infdis/jis718.
77. Wong SM, Akerley BJ. 2005. Environmental and genetic regulation of the phosphorylcholine epitope of Haemophilus influenzae lipooligosaccharide. Mol. Microbiol. 55:724-738. http://dx.doi.org/10.1111/j.1365-2958 .2004.04439.x.