Abstract
As a step toward developing recombinant oral vaccines, we have explored the feasibility of expression of O polysaccharide antigens from Pseudomonas aeruginosa by Escherichia coli. We cloned in E. coli HB101 a 26.2-kilobase DNA fragment from P. aeruginosa strain PA103 that specifies the production of the O polysaccharide of Fisher immunotype 2 (IT-2) strains. The recombinant organism incorporated the P. aeruginosa IT-2 O polysaccharide onto the core of the E. coli lipopolysaccharide (LPS). Transfer of the recombinant plasmid to three LPS-rough strains of P. aeruginosa resulted in synthesis of IT-2 O antigen, and two of these transconjugant strains also synthesized a second O polysaccharide, presumably representing expression of a repressed, or an incomplete, set of genes for an endogenous O polysaccharide. Rabbits injected with the purified recombinant LPS made antibody specific for P. aeruginosa IT-2 O side chains, as did mice fed the recombinant E. coli strain. Expression of P. aeruginosa O antigens by enteric bacteria makes it possible to study these recombinant strains as oral vaccines to prevent P. aeruginosa infections.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abraham E., Robinson A. Oral immunization with bacterial polysaccharide and adjuvant enhances antigen-specific pulmonary secretory antibody response and resistance to pneumonia. Vaccine. 1991 Oct;9(10):757–764. doi: 10.1016/0264-410x(91)90293-f. [DOI] [PubMed] [Google Scholar]
- Bachmann B. J. Linkage map of Escherichia coli K-12, edition 8. Microbiol Rev. 1990 Jun;54(2):130–197. doi: 10.1128/mr.54.2.130-197.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baron L. S., Kopecko D. J., Formal S. B., Seid R., Guerry P., Powell C. Introduction of Shigella flexneri 2a type and group antigen genes into oral typhoid vaccine strain Salmonella typhi Ty21a. Infect Immun. 1987 Nov;55(11):2797–2801. doi: 10.1128/iai.55.11.2797-2801.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bhat R., Marx A., Galanos C., Conrad R. S. Structural studies of lipid A from Pseudomonas aeruginosa PAO1: occurrence of 4-amino-4-deoxyarabinose. J Bacteriol. 1990 Dec;172(12):6631–6636. doi: 10.1128/jb.172.12.6631-6636.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
- Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Curtiss R., 3rd, Kelly S. M. Salmonella typhimurium deletion mutants lacking adenylate cyclase and cyclic AMP receptor protein are avirulent and immunogenic. Infect Immun. 1987 Dec;55(12):3035–3043. doi: 10.1128/iai.55.12.3035-3043.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Engels W., Endert J., Kamps M. A., van Boven C. P. Role of lipopolysaccharide in opsonization and phagocytosis of Pseudomonas aeruginosa. Infect Immun. 1985 Jul;49(1):182–189. doi: 10.1128/iai.49.1.182-189.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fegan M., Francis P., Hayward A. C., Davis G. H., Fuerst J. A. Phenotypic conversion of Pseudomonas aeruginosa in cystic fibrosis. J Clin Microbiol. 1990 Jun;28(6):1143–1146. doi: 10.1128/jcm.28.6.1143-1146.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fisher M. W., Devlin H. B., Gnabasik F. J. New immunotype schema for Pseudomonas aeruginosa based on protective antigens. J Bacteriol. 1969 May;98(2):835–836. doi: 10.1128/jb.98.2.835-836.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forrest B. D., LaBrooy J. T. In vivo evidence of immunological masking of the Vibrio cholerae O antigen of a hybrid Salmonella typhi Ty21a-Vibrio cholerae oral vaccine in humans. Vaccine. 1991 Jul;9(7):515–520. doi: 10.1016/0264-410x(91)90039-9. [DOI] [PubMed] [Google Scholar]
- Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
- Goldberg J. B., Ohman D. E. Cloning and expression in Pseudomonas aeruginosa of a gene involved in the production of alginate. J Bacteriol. 1984 Jun;158(3):1115–1121. doi: 10.1128/jb.158.3.1115-1121.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hooke A. M., Cerquetti M. C., Wan K. S., Wang Z., Sordelli D. O., Bellanti J. A. Oral immunization of mice with temperature-sensitive Pseudomonas aeruginosa enhances pulmonary clearance of the wild-type. Vaccine. 1991 May;9(5):294–296. doi: 10.1016/0264-410x(91)90052-8. [DOI] [PubMed] [Google Scholar]
- Knirel YuA, Vinogradov E. V., Kocharova N. A., Paramonov N. A., Kochetkov N. K., Dmitriev B. A., Stanislavsky E. S., Lányi B. The structure of O-specific polysaccharides and serological classification of Pseudomonas aeruginosa (a review). Acta Microbiol Hung. 1988;35(1):3–24. [PubMed] [Google Scholar]
- Knirel Y. A. Polysaccharide antigens of Pseudomonas aeruginosa. Crit Rev Microbiol. 1990;17(4):273–304. doi: 10.3109/10408419009105729. [DOI] [PubMed] [Google Scholar]
- Kocharova N. A., Knirel Y. A., Shashkov A. S., Kochetkov N. K., Pier G. B. Structure of an extracellular cross-reactive polysaccharide from Pseudomonas aeruginosa immunotype 4. J Biol Chem. 1988 Aug 15;263(23):11291–11295. [PubMed] [Google Scholar]
- Liu P. V. Exotoxins of Pseudomonas aeruginosa. I. Factors that influence the production of exotoxin A. J Infect Dis. 1973 Oct;128(4):506–513. doi: 10.1093/infdis/128.4.506. [DOI] [PubMed] [Google Scholar]
- Penketh A., Pitt T., Roberts D., Hodson M. E., Batten J. C. The relationship of phenotype changes in Pseudomonas aeruginosa to the clinical condition of patients with cystic fibrosis. Am Rev Respir Dis. 1983 May;127(5):605–608. doi: 10.1164/arrd.1983.127.5.605. [DOI] [PubMed] [Google Scholar]
- Pier G. B., Ames P. Mediation of the killing of rough, mucoid isolates of Pseudomonas aeruginosa from patients with cystic fibrosis by the alternative pathway of complement. J Infect Dis. 1984 Aug;150(2):223–228. doi: 10.1093/infdis/150.2.223. [DOI] [PubMed] [Google Scholar]
- Pier G. B. Polysaccharide antigens of Pseudomonas aeruginosa. Rev Infect Dis. 1988 Jul-Aug;10 (Suppl 2):S337–S340. doi: 10.1093/cid/10.supplement_2.s337. [DOI] [PubMed] [Google Scholar]
- Pier G. B., Sidberry H. F., Zolyomi S., Sadoff J. C. Isolation and characterization of a high-molecular-weight polysaccharide from the slime of Pseudomonas aeruginosa. Infect Immun. 1978 Dec;22(3):908–918. doi: 10.1128/iai.22.3.908-918.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pier G. B., Thomas D. M. Lipopolysaccharide and high-molecular-weight polysaccharide serotypes of Pseudomonas aeruginosa. J Infect Dis. 1982 Feb;145(2):217–223. doi: 10.1093/infdis/145.2.217. [DOI] [PubMed] [Google Scholar]
- Pitt T. L., MacDougall J., Penketh A. R., Cooke E. M. Polyagglutinating and non-typable strains of Pseudomonas aeruginosa in cystic fibrosis. J Med Microbiol. 1986 Mar;21(2):179–186. doi: 10.1099/00222615-21-2-179. [DOI] [PubMed] [Google Scholar]
- Pollack M., Young L. S. Protective activity of antibodies to exotoxin A and lipopolysaccharide at the onset of Pseudomonas aeruginosa septicemia in man. J Clin Invest. 1979 Feb;63(2):276–286. doi: 10.1172/JCI109300. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Sanderson K. E., Roth J. R. Linkage map of Salmonella typhimurium, edition VII. Microbiol Rev. 1988 Dec;52(4):485–532. doi: 10.1128/mr.52.4.485-532.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seid R. C., Jr, Kopecko D. J., Sadoff J. C., Schneider H., Baron L. S., Formal S. B. Unusual lipopolysaccharide antigens of a Salmonella typhi oral vaccine strain expressing the Shigella sonnei form I antigen. J Biol Chem. 1984 Jul 25;259(14):9028–9034. [PubMed] [Google Scholar]
- Tacket C. O., Hone D. M., Curtiss R., 3rd, Kelly S. M., Losonsky G., Guers L., Harris A. M., Edelman R., Levine M. M. Comparison of the safety and immunogenicity of delta aroC delta aroD and delta cya delta crp Salmonella typhi strains in adult volunteers. Infect Immun. 1992 Feb;60(2):536–541. doi: 10.1128/iai.60.2.536-541.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young L. S. Human immunity to Pseudomonas aeruginosa. II. Relationship between heat-stable opsonins and type-specific lipopolysaccharides. J Infect Dis. 1972 Sep;126(3):277–287. doi: 10.1093/infdis/126.3.277. [DOI] [PubMed] [Google Scholar]
- Young L. S. Role of antibody in infections due to Pseudomonas aeruginosa. J Infect Dis. 1974 Nov;130 (Suppl)(0):S111–S118. doi: 10.1093/infdis/130.supplement.s111. [DOI] [PubMed] [Google Scholar]
- Young L. S., Yu B. H., Armstrong D. Agar-Gel Precipitating Antibody in Pseudomonas aeruginosa Infections. Infect Immun. 1970 Oct;2(4):495–503. doi: 10.1128/iai.2.4.495-503.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]