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. 1987 Nov;25(11):2080–2084. doi: 10.1128/jcm.25.11.2080-2084.1987

Purified 60-kilodalton Legionella protein antigen with Legionella-specific and nonspecific epitopes.

B B Plikaytis 1, G M Carlone 1, C P Pau 1, H W Wilkinson 1
PMCID: PMC269415  PMID: 2447117

Abstract

In a previous study, all convalescent-phase sera from patients with culture-confirmed legionellosis reacted on immunoblots with a Legionella genus-wide 58-kilodalton (kDa) protein antigen (J.S. Sampson, B.B. Plikaytis, and H.W. Wilkinson, J. Clin. Microbiol. 23:92-99, 1986). The present study was done to immunologically characterize and determine the diagnostic relevance of this purified antigen. The antigen was precipitated from enriched cell extracts with ammonium sulfate and purified by high-pressure liquid chromatography. High-titered rabbit antiserum produced to the purified protein was used to show its presence on immunoblots in the 60-kDa range in 38 Legionella serogroups, representing 23 species, and in 39 non-Legionella bacteria. The antiserum was made specific for Legionella strains by sequential absorptions with Bordetella pertussis, Pseudomonas aeruginosa, and Pseudomonas fluorescens whole cells. Serum from legionellosis patients reacted with both specific and nonspecific epitopes. Results of indirect immunofluorescence experiments showed that neither specific nor nonspecific epitopes of the 60-kDa protein were surface exposed on Legionella cells and that cross-reactive epitopes were variably exposed on non-Legionella bacteria. The 60-kDa protein antigen should be useful in diagnostic tests for legionellosis if care is taken to expose cryptic epitopes and if the tests use or measure only the Legionella-specific epitopes.

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Selected References

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

  1. Carlone G. M., Plikaytis B. B., Arko R. J. Immune serum to protein molecular weight standards for calibrating Western blots. Anal Biochem. 1986 May 15;155(1):89–91. doi: 10.1016/0003-2697(86)90229-0. [DOI] [PubMed] [Google Scholar]
  2. Collins M. T., Espersen F., Høiby N., Cho S. N., Friis-Møller A., Reif J. S. Cross-reactions between Legionella pneumophila (serogroup 1) and twenty-eight other bacterial species, including other members of the family Legionellaceae. Infect Immun. 1983 Mar;39(3):1441–1456. doi: 10.1128/iai.39.3.1441-1456.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hindersson P., Knudsen J. D., Axelsen N. H. Cloning and expression of treponema pallidum common antigen (Tp-4) in Escherichia coli K12. J Gen Microbiol. 1987 Mar;133(3):587–596. doi: 10.1099/00221287-133-3-587. [DOI] [PubMed] [Google Scholar]
  4. Hindersson P., Petersen C. S., Pedersen N. S., Høiby N., Axelsen N. H. Immunological cross-reaction between antigen Tp-4 of Treponema pallidum and an antigen common to a wide range of bacteria. Acta Pathol Microbiol Immunol Scand B. 1984 Aug;92(4):183–188. doi: 10.1111/j.1699-0463.1984.tb02818.x. [DOI] [PubMed] [Google Scholar]
  5. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  6. Plikaytis B. D., Carlone G. M., Edmonds P., Mayer L. W. Robust estimation of standard curves for protein molecular weight and linear-duplex DNA base-pair number after gel electrophoresis. Anal Biochem. 1986 Feb 1;152(2):346–364. doi: 10.1016/0003-2697(86)90420-3. [DOI] [PubMed] [Google Scholar]
  7. Plikaytis B. D., Carlone G. M., Plikaytis B. B. Numerical analysis of normalized whole-cell protein profiles after sodium dodecyl sulphate-polyacrylamide gel electrophoresis. J Gen Microbiol. 1986 Sep;132(9):2653–2660. doi: 10.1099/00221287-132-9-2653. [DOI] [PubMed] [Google Scholar]
  8. Sampson J. S., Plikaytis B. B., Wilkinson H. W. Immunologic response of patients with legionellosis against major protein-containing antigens of Legionella pneumophila serogroup 1 as shown by immunoblot analysis. J Clin Microbiol. 1986 Jan;23(1):92–99. doi: 10.1128/jcm.23.1.92-99.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Sompolinsky D., Hertz J. B., Høiby N., Jensen K., Mansa B., Pedersen V. B., Samra Z. An antigen common to a wide range of bacteria. 2. A biochemical study of a "common antigen" from Pseudomonas aeruginosa. Acta Pathol Microbiol Scand B. 1980 Oct;88(5):253–260. doi: 10.1111/j.1699-0463.1980.tb02637.x. [DOI] [PubMed] [Google Scholar]
  10. Sompolinsky D., Hertz J. B., Høiby N., Jensen K., Mansa B., Samra Z. An antigen common to a wide range of bacteria. I. The isolation of a 'common antigen' from Pseudomonas aeruginosa. Acta Pathol Microbiol Scand B. 1980 Jun;88(3):143–149. doi: 10.1111/j.1699-0463.1980.tb02620.x. [DOI] [PubMed] [Google Scholar]
  11. Tsang V. C., Hancock K., Simons A. R. Calibration of prestained protein molecular weight standards for use in the "Western" or enzyme-linked immunoelectrotransfer blot techniques. Anal Biochem. 1984 Dec;143(2):304–307. doi: 10.1016/0003-2697(84)90667-5. [DOI] [PubMed] [Google Scholar]

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