Skip to main content
PMC home page
Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1995 Mar;2(2):160–165. doi: 10.1128/cdli.2.2.160-165.1995

Monoclonal antibodies to Legionella Mip proteins recognize genus- and species-specific epitopes.

J H Helbig 1, B Ludwig 1, P C Lück 1, A Groh 1, W Witzleb 1, J Hacker 1
PMCID: PMC170120  PMID: 7535177

Abstract

Monoclonal antibodies (MAbs) against the virulence-associated Mip protein of Legionella spp. were raised by immunizing BALB/c mice with (i) Legionella pneumophila, (ii) Legionella micdadei, and (iii) purified recombinant native Mip protein cloned from L. pneumophila Philadelphia 1. Following screening of seeded wells by immunoblot analysis with homologous antigens, eight Mip-specific MAbs were found. These MAbs were chosen to investigate the antigenic diversity of Mip proteins in the genus Legionella. Mip was detected in 82 Legionella strains representing all 34 species tested. One of these MAbs, obtained from immunization with L. micdadei, recognized an epitope common to all Legionella species tested by immunoblot analysis. Another MAb was discovered to be specific for the Mip protein of L. pneumophila. The remaining six MAbs recognized 18 to 79% of Legionella species included in this study. By making use of the MAbs introduced in this study, it could be shown that, based on Mip protein epitope expression, Legionella species can be divided into at least six antigenetically distinct groups. As demonstrated by 43 L. pneumophila strains representing all serogroups, no antigenic diversity of Mip proteins was found for this species. In addition, 18 non-Legionella species, including Chlamydia trachomatis, Neisseria meningitidis, Pseudomonas aeruginosa, and Saccharomyces cerevisiae, all of which are known to carry genes homologous to the Legionella mip genes, were reacted against all eight MAbs. No cross-reactivity was detectable in any of those strains.

Full Text

The Full Text of this article is available as a PDF (413.6 KB).

Selected References

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

  1. Bangsborg J. M., Cianciotto N. P., Hindersson P. Nucleotide sequence analysis of the Legionella micdadei mip gene, encoding a 30-kilodalton analog of the Legionella pneumophila Mip protein. Infect Immun. 1991 Oct;59(10):3836–3840. doi: 10.1128/iai.59.10.3836-3840.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bender L., Ott M., Marre R., Hacker J. Genome analysis of Legionella ssp. by orthogonal field alternation gel electrophoresis (OFAGE). FEMS Microbiol Lett. 1990 Nov;60(3):253–257. doi: 10.1016/0378-1097(90)90313-f. [DOI] [PubMed] [Google Scholar]
  3. Cianciotto N. P., Bangsborg J. M., Eisenstein B. I., Engleberg N. C. Identification of mip-like genes in the genus Legionella. Infect Immun. 1990 Sep;58(9):2912–2918. doi: 10.1128/iai.58.9.2912-2918.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cianciotto N. P., Eisenstein B. I., Mody C. H., Engleberg N. C. A mutation in the mip gene results in an attenuation of Legionella pneumophila virulence. J Infect Dis. 1990 Jul;162(1):121–126. doi: 10.1093/infdis/162.1.121. [DOI] [PubMed] [Google Scholar]
  5. Cianciotto N. P., Eisenstein B. I., Mody C. H., Toews G. B., Engleberg N. C. A Legionella pneumophila gene encoding a species-specific surface protein potentiates initiation of intracellular infection. Infect Immun. 1989 Apr;57(4):1255–1262. doi: 10.1128/iai.57.4.1255-1262.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cianciotto N. P., Fields B. S. Legionella pneumophila mip gene potentiates intracellular infection of protozoa and human macrophages. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5188–5191. doi: 10.1073/pnas.89.11.5188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cianfriglia M., Armellini D., Massone A., Mariani M. Simple immunization protocol for high frequency production of soluble antigen-specific hybridomas. Hybridoma. 1983;2(4):451–457. doi: 10.1089/hyb.1983.2.451. [DOI] [PubMed] [Google Scholar]
  8. Dennis P. J., Brenner D. J., Thacker W. L., Wait R., Vesey G., Steigerwalt A. G., Benson R. F. Five new Legionella species isolated from water. Int J Syst Bacteriol. 1993 Apr;43(2):329–337. doi: 10.1099/00207713-43-2-329. [DOI] [PubMed] [Google Scholar]
  9. Dowling J. N., Saha A. K., Glew R. H. Virulence factors of the family Legionellaceae. Microbiol Rev. 1992 Mar;56(1):32–60. doi: 10.1128/mr.56.1.32-60.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Edelstein P. H. Comparative study of selective media for isolation of Legionella pneumophila from potable water. J Clin Microbiol. 1982 Oct;16(4):697–699. doi: 10.1128/jcm.16.4.697-699.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fischer G., Bang H., Ludwig B., Mann K., Hacker J. Mip protein of Legionella pneumophila exhibits peptidyl-prolyl-cis/trans isomerase (PPlase) activity. Mol Microbiol. 1992 May;6(10):1375–1383. doi: 10.1111/j.1365-2958.1992.tb00858.x. [DOI] [PubMed] [Google Scholar]
  12. Hacker J., Fischer G. Immunophilins: structure-function relationship and possible role in microbial pathogenicity. Mol Microbiol. 1993 Nov;10(3):445–456. doi: 10.1111/j.1365-2958.1993.tb00917.x. [DOI] [PubMed] [Google Scholar]
  13. Jepras R. I., Fitzgeorge R. B., Baskerville A. A comparison of virulence of two strains of Legionella pneumophila based on experimental aerosol infection of guinea-pigs. J Hyg (Lond) 1985 Aug;95(1):29–38. doi: 10.1017/s0022172400062252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lever M. S. Production and characterisation of a Legionella pneumophila specific monoclonal antibody. FEMS Microbiol Lett. 1993 Feb 15;107(1):5–9. doi: 10.1016/0378-1097(93)90345-3. [DOI] [PubMed] [Google Scholar]
  16. Ludwig B., Rahfeld J., Schmidt B., Mann K., Wintermeyer E., Fischer G., Hacker J. Characterization of Mip proteins of Legionella pneumophila. FEMS Microbiol Lett. 1994 May 1;118(1-2):23–30. doi: 10.1111/j.1574-6968.1994.tb06798.x. [DOI] [PubMed] [Google Scholar]
  17. Lundemose A. G., Birkelund S., Fey S. J., Larsen P. M., Christiansen G. Chlamydia trachomatis contains a protein similar to the Legionella pneumophila mip gene product. Mol Microbiol. 1991 Jan;5(1):109–115. doi: 10.1111/j.1365-2958.1991.tb01831.x. [DOI] [PubMed] [Google Scholar]
  18. Nolte F. S., Conlin C. A. Major outer membrane protein of Legionella pneumophila carries a species-specific epitope. J Clin Microbiol. 1986 Mar;23(3):643–646. doi: 10.1128/jcm.23.3.643-646.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Peters H., Jürs M., Jann B., Jann K., Timmis K. N., Bitter-Suermann D. Monoclonal antibodies to enterobacterial common antigen and to Escherichia coli lipopolysaccharide outer core: demonstration of an antigenic determinant shared by enterobacterial common antigen and E. coli K5 capsular polysaccharide. Infect Immun. 1985 Nov;50(2):459–466. doi: 10.1128/iai.50.2.459-466.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rowbotham T. J. Current views on the relationships between amoebae, legionellae and man. Isr J Med Sci. 1986 Sep;22(9):678–689. [PubMed] [Google Scholar]
  21. Steinmetz I., Rheinheimer C., Hübner I., Bitter-Suermann D. Genus-specific epitope on the 60-kilodalton Legionella heat shock protein recognized by a monoclonal antibody. J Clin Microbiol. 1991 Feb;29(2):346–354. doi: 10.1128/jcm.29.2.346-354.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Clinical and Diagnostic Laboratory Immunology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES