Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Oct;57(10):3045–3052. doi: 10.1128/iai.57.10.3045-3052.1989

Identification of a chromosomal locus for expression of lipopolysaccharide epitopes in Haemophilus influenzae.

J N Weiser 1, A A Lindberg 1, E J Manning 1, E J Hansen 1, E R Moxon 1
PMCID: PMC260768  PMID: 2476397

Abstract

Lipopolysaccharide (LPS) is a major virulence determinant of Haemophilus influenzae. The organism is able to display an extensive repertoire of different LPS structures through the loss and acquisition of multiple oligosaccharide epitopes in various combinations. This marked heterogeneity of LPS molecules has complicated the analysis of the structure of LPS and its role in pathogenesis. A genomic library was screened for the ability to transform H. influenzae to express novel LPS epitopes defined by reactivity with oligosaccharide specific monoclonal antibodies. A chromosomal locus, lic-1, involved in expression of at least three different epitopes (recognized by monoclonal antibodies 4C4, 12D9, and 6A2), was identified on a 5.6-kilobase restriction endonuclease fragment. Transformation of H. influenzae with subclones from within lic-1 was used to generate a series of isogenic and phenotypic variants. All transformants displayed phase variation for their newly acquired epitopes. Altered binding specificities of LPS with monoclonal antibodies correlated with changes in sugar compositional analysis. The expression of two epitopes was eliminated by introduction of site-specific mutations in lic-1, confirming the role of lic-1 in oligosaccharide biosynthesis.

Full text

PDF
3046

Images in this article

3048Image
on p.3048
3049Image
on p.3049
3050Image
on p.3050
3050Image
on p.3050

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cryz S. J., Jr, Pitt T. L., Fürer E., Germanier R. Role of lipopolysaccharide in virulence of Pseudomonas aeruginosa. Infect Immun. 1984 May;44(2):508–513. doi: 10.1128/iai.44.2.508-513.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  3. Griffiss J. M., Schneider H., Mandrell R. E., Yamasaki R., Jarvis G. A., Kim J. J., Gibson B. W., Hamadeh R., Apicella M. A. Lipooligosaccharides: the principal glycolipids of the neisserial outer membrane. Rev Infect Dis. 1988 Jul-Aug;10 (Suppl 2):S287–S295. doi: 10.1093/cid/10.supplement_2.s287. [DOI] [PubMed] [Google Scholar]
  4. Gulig P. A., Frisch C. F., Hansen E. J. A set of two monoclonal antibodies specific for the cell surface-exposed 39K major outer membrane protein of Haemophilus influenzae type b defines all strains of this pathogen. Infect Immun. 1983 Nov;42(2):516–524. doi: 10.1128/iai.42.2.516-524.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gulig P. A., Patrick C. C., Hermanstorfer L., McCracken G. H., Jr, Hansen E. J. Conservation of epitopes in the oligosaccharide portion of the lipooligosaccharide of Haemophilus influenzae type b. Infect Immun. 1987 Mar;55(3):513–520. doi: 10.1128/iai.55.3.513-520.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Herriott R. M., Meyer E. M., Vogt M. Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol. 1970 Feb;101(2):517–524. doi: 10.1128/jb.101.2.517-524.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hoiseth S. K., Connelly C. J., Moxon E. R. Genetics of spontaneous, high-frequency loss of b capsule expression in Haemophilus influenzae. Infect Immun. 1985 Aug;49(2):389–395. doi: 10.1128/iai.49.2.389-395.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Inzana T. J. Electrophoretic heterogeneity and interstrain variation of the lipopolysaccharide of Haemophilus influenzae. J Infect Dis. 1983 Sep;148(3):492–499. doi: 10.1093/infdis/148.3.492. [DOI] [PubMed] [Google Scholar]
  9. Inzana T. J., Seifert W. E., Jr, Williams R. P. Composition and antigenic activity of the oligosaccharide moiety of Haemophilus influenzae type b lipooligosaccharide. Infect Immun. 1985 May;48(2):324–330. doi: 10.1128/iai.48.2.324-330.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kadam S. K., Rehemtulla A., Sanderson K. E. Cloning of rfaG, B, I, and J genes for glycosyltransferase enzymes for synthesis of the lipopolysaccharide core of Salmonella typhimurium. J Bacteriol. 1985 Jan;161(1):277–284. doi: 10.1128/jb.161.1.277-284.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kimura A., Hansen E. J. Antigenic and phenotypic variations of Haemophilus influenzae type b lipopolysaccharide and their relationship to virulence. Infect Immun. 1986 Jan;51(1):69–79. doi: 10.1128/iai.51.1.69-79.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kimura A., Patrick C. C., Miller E. E., Cope L. D., McCracken G. H., Jr, Hansen E. J. Haemophilus influenzae type b lipooligosaccharide: stability of expression and association with virulence. Infect Immun. 1987 Sep;55(9):1979–1986. doi: 10.1128/iai.55.9.1979-1986.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lee J. J., Smith H. O., Redfield R. J. Organization of the Haemophilus influenzae Rd genome. J Bacteriol. 1989 Jun;171(6):3016–3024. doi: 10.1128/jb.171.6.3016-3024.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pluschke G., Mercer A., Kusećek B., Pohl A., Achtman M. Induction of bacteremia in newborn rats by Escherichia coli K1 is correlated with only certain O (lipopolysaccharide) antigen types. Infect Immun. 1983 Feb;39(2):599–608. doi: 10.1128/iai.39.2.599-608.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Smith H. O., Danner D. B., Deich R. A. Genetic transformation. Annu Rev Biochem. 1981;50:41–68. doi: 10.1146/annurev.bi.50.070181.000353. [DOI] [PubMed] [Google Scholar]
  16. Tolan R. W., Jr, Munson R. S., Jr, Granoff D. M. Lipopolysaccharide gel profiles of Haemophilus influenzae type b are not stable epidemiologic markers. J Clin Microbiol. 1986 Aug;24(2):223–227. doi: 10.1128/jcm.24.2.223-227.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Zamze S. E., Ferguson M. A., Moxon E. R., Dwek R. A., Rademacher T. W. Identification of phosphorylated 3-deoxy-manno-octulosonic acid as a component of Haemophilus influenzae lipopolysaccharide. Biochem J. 1987 Jul 15;245(2):583–587. doi: 10.1042/bj2450583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Zamze S. E., Moxon E. R. Composition of the lipopolysaccharide from different capsular serotype strains of Haemophilus influenzae. J Gen Microbiol. 1987 Jun;133(6):1443–1451. doi: 10.1099/00221287-133-6-1443. [DOI] [PubMed] [Google Scholar]
  19. Zwahlen A., Rubin L. G., Connelly C. J., Inzana T. J., Moxon E. R. Alteration of the cell wall of Haemophilus influenzae type b by transformation with cloned DNA: association with attenuated virulence. J Infect Dis. 1985 Sep;152(3):485–492. doi: 10.1093/infdis/152.3.485. [DOI] [PubMed] [Google Scholar]
  20. Zwahlen A., Rubin L. G., Moxon E. R. Contribution of lipopolysaccharide to pathogenicity of Haemophilus influenzae: comparative virulence of genetically-related strains in rats. Microb Pathog. 1986 Oct;1(5):465–473. doi: 10.1016/0882-4010(86)90008-2. [DOI] [PubMed] [Google Scholar]
  21. van Alphen L., Riemens T., Poolman J., Hopman C., Zanen H. C. Homogeneity of cell envelope protein subtypes, lipopolysaccharide serotypes, and biotypes among Haemophilus influenzae type b from patients with meningitis in The Netherlands. J Infect Dis. 1983 Jul;148(1):75–81. doi: 10.1093/infdis/148.1.75. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES