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. 1995 Nov;2(6):747–752. doi: 10.1128/cdli.2.6.747-752.1995

Identification of outer membrane proteins as target antigens of Pseudomonas aeruginosa Homma serotype M.

S Yokota 1
PMCID: PMC170232  PMID: 8574841

Abstract

Pseudomonas aeruginosa is routinely serotyped in Japan by using the Homma scheme. The serotypes (O serotypes) are based on the chemical structure of the O-polysaccharide portion of the lipopolysaccharide (LPS). However, the nature of the Homma serotype M antigen has remained obscure because strains classified as serotype M usually have the rough phenotype. I characterized the target antigen of serotype M. The results of Western blotting (immunoblotting) showed that commercially available typing monoclonal antibody (MAb) against serotype M specifically bound to outer membrane protein (Opr) G and that typing rabbit antiserum specific for serotype M mainly contained antibodies against Oprs F and H2. These Oprs were distributed among all P. aeruginosa strains tested, including the serotype standard, serotype M and nontypeable strains, and a series of LPS-core-defective mutants derived from strain PAC1. However, the rough mutants derived from strain PAC1 agglutinated with the anti-serotype M antibodies, whereas the smooth strains did not. LPS preparations from serotype M strains possessed few or no polysaccharide chains. These strains had higher levels of binding activity with anti-serotype M MAb, as well as with anti-lipid A MAb, which specifically bound to the cell surface of the rough-natured gram-negative bacterial strains with high activity. The anti-serotype M antiserum also contained rough-LPS-specific antibodies, but the epitope was distributed among only a few strains. The results suggested that the Oprs acted as the serotype M antigen and that LPS did not. In conclusion, the rough strains agglutinated with anti-Opr antibodies and were distinguished as serotype M from the smooth strains of other serotypes, because the antibodies were accessible to the cell surface lacking O polysaccharides. I supposed that Homma serotype M is an index of the rough nature of P. aeruginosa strains rather than one of the O serotypes.

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Selected References

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  1. Fomsgaard A., Conrad R. S., Galanos C., Shand G. H., Høiby N. Comparative immunochemistry of lipopolysaccharides from typable and polyagglutinable Pseudomonas aeruginosa strains isolated from patients with cystic fibrosis. J Clin Microbiol. 1988 May;26(5):821–826. doi: 10.1128/jcm.26.5.821-826.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Garin-Bastuji B., Bowden R. A., Dubray G., Limet J. N. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis of smooth-lipopolysaccharide heterogeneity among Brucella biovars related to A and M specificities. J Clin Microbiol. 1990 Oct;28(10):2169–2174. doi: 10.1128/jcm.28.10.2169-2174.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hancock R. E., Carey A. M. Outer membrane of Pseudomonas aeruginosa: heat- 2-mercaptoethanol-modifiable proteins. J Bacteriol. 1979 Dec;140(3):902–910. doi: 10.1128/jb.140.3.902-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hancock R. E., Mouat E. C., Speert D. P. Quantitation and identification of antibodies to outer-membrane proteins of Pseudomonas aeruginosa in sera of patients with cystic fibrosis. J Infect Dis. 1984 Feb;149(2):220–226. doi: 10.1093/infdis/149.2.220. [DOI] [PubMed] [Google Scholar]
  5. Hancock R. E., Mutharia L. M., Chan L., Darveau R. P., Speert D. P., Pier G. B. Pseudomonas aeruginosa isolates from patients with cystic fibrosis: a class of serum-sensitive, nontypable strains deficient in lipopolysaccharide O side chains. Infect Immun. 1983 Oct;42(1):170–177. doi: 10.1128/iai.42.1.170-177.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hofstra H., Dankert J. Major outer membrane proteins: common antigens in enterobacteriaceae species. J Gen Microbiol. 1980 Jul;119(1):123–131. doi: 10.1099/00221287-119-1-123. [DOI] [PubMed] [Google Scholar]
  7. Homma J. Y. Designation of the thirteen O-group antigens of Pseudomonas aeruginosa; an amendment for the tentative proposal in 1976. Jpn J Exp Med. 1982 Dec;52(6):317–320. [PubMed] [Google Scholar]
  8. Knirel Y. A. Polysaccharide antigens of Pseudomonas aeruginosa. Crit Rev Microbiol. 1990;17(4):273–304. doi: 10.3109/10408419009105729. [DOI] [PubMed] [Google Scholar]
  9. Kobayashi I., Hasegawa M., Miyazaki S., Nishida M., Goto S. In vitro and in vivo changes of serotype in Pseudomonas aeruginosa isolates by anti-pseudomonal drugs. J Antibiot (Tokyo) 1994 Jan;47(1):72–79. doi: 10.7164/antibiotics.47.72. [DOI] [PubMed] [Google Scholar]
  10. Kobayashi I., Hasegawa M., Nishida M. [Formation of drug-induced serotype variants of Pseudomonas aeruginosa: changes in biochemical properties, drug susceptibility and outer membrane proteins]. Kansenshogaku Zasshi. 1994 Feb;68(2):183–190. doi: 10.11150/kansenshogakuzasshi1970.68.183. [DOI] [PubMed] [Google Scholar]
  11. Lam M. Y., McGroarty E. J., Kropinski A. M., MacDonald L. A., Pedersen S. S., Høiby N., Lam J. S. Occurrence of a common lipopolysaccharide antigen in standard and clinical strains of Pseudomonas aeruginosa. J Clin Microbiol. 1989 May;27(5):962–967. doi: 10.1128/jcm.27.5.962-967.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meadow P. M., Rowe P. S., Wells P. L. Characterization of polyagglutinating and surface antigens in Pseudomonas aeruginosa. J Gen Microbiol. 1984 Mar;130(3):631–644. doi: 10.1099/00221287-130-3-631. [DOI] [PubMed] [Google Scholar]
  13. Mutharia L. M., Nicas T. I., Hancock R. E. Outer membrane proteins of Pseudomonas aeruginosa serotype strains. J Infect Dis. 1982 Dec;146(6):770–779. doi: 10.1093/infdis/146.6.770. [DOI] [PubMed] [Google Scholar]
  14. Ojeniyi B., Baek L., Høiby N. Polyagglutinability due to loss of O-antigenic determinants in Pseudomonas aeruginosa strains isolated from cystic fibrosis patients. Acta Pathol Microbiol Immunol Scand B. 1985 Feb;93(1):7–13. doi: 10.1111/j.1699-0463.1985.tb02844.x. [DOI] [PubMed] [Google Scholar]
  15. Rowe P. S., Meadow P. M. Structure of the Core oligosaccharide from the lipopolysaccharide of Pseudomonas aeruginosa PAC1R and its defective mutants. Eur J Biochem. 1983 May 2;132(2):329–337. doi: 10.1111/j.1432-1033.1983.tb07366.x. [DOI] [PubMed] [Google Scholar]
  16. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  17. Strickland M. A., Gaston M. A., Pitt T. L. Comparison of polyclonal rabbit antisera with monoclonal antibodies for serological typing of Pseudomonas aeruginosa. J Clin Microbiol. 1988 Apr;26(4):768–769. doi: 10.1128/jcm.26.4.768-769.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  19. Yokota S., Hayashi T., Matsumoto H. Identification of the lipopolysaccharide core region as the receptor site for a cytotoxin-converting phage, phi CTX, of Pseudomonas aeruginosa. J Bacteriol. 1994 Sep;176(17):5262–5269. doi: 10.1128/jb.176.17.5262-5269.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yokota S., Kaya S., Araki Y., Ito E., Kawamura T., Sawada S. Occurrence of D-rhamnan as the common antigen reactive against monoclonal antibody E87 in Pseudomonas aeruginosa IFO 3080 and other strains. J Bacteriol. 1990 Oct;172(10):6162–6164. doi: 10.1128/jb.172.10.6162-6164.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yokota S., Ochi H., Ohtsuka H., Kato M., Noguchi H. Heterogeneity of the L-rhamnose residue in the outer core of Pseudomonas aeruginosa lipopolysaccharide, characterized by using human monoclonal antibodies. Infect Immun. 1989 Jun;57(6):1691–1696. doi: 10.1128/iai.57.6.1691-1696.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yokota S., Terashima M., Chiba J., Noguchi H. Variable cross-reactivity of Pseudomonas aeruginosa lipopolysaccharide-code-specific monoclonal antibodies and its possible relationship with serotype. J Gen Microbiol. 1992 Feb;138(2):289–296. doi: 10.1099/00221287-138-2-289. [DOI] [PubMed] [Google Scholar]
  23. Yoshida K., Kimura F., Uchida K., Kawaharajo K., Koizumi N., Nakajima S., Nagaoka K. Production and some properties of monoclonal antibodies against serotype strains of Pseudomonas aeruginosa. J Pharmacobiodyn. 1989 Jul;12(7):398–404. doi: 10.1248/bpb1978.12.398. [DOI] [PubMed] [Google Scholar]

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