microbiology, Vol 140, 2003-2011, Copyright © 1994 by Society for General Microbiology

ARTICLES

Immunoreactivity of the 60 kDa cysteine-rich proteins of Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae expressed in Escherichia coli

MW Watson, PR Lambden, JS Everson and IN Clarke
Faculty of Medicine, Southampton General Hospital, UK.

The 60 kDa cysteine-rich proteins (CrPs) of Chlamydia are developmentally regulated outer envelope proteins synthesized late in the chlamydial growth cycle. These proteins, found only on the extracellular infectious elementary bodies, elicit major antibody responses in chlamydial infection. We have cloned and expressed in Escherichia coli the complete 60 kDa CrP genes from Chlamydia trachomatis, C. psittaci and C. pneumoniae. The recombinant products were expressed as either 'native' proteins or as fusions with the bacteriophage T7 gene 10 protein. Electron microscopy showed that recombinant proteins were produced as insoluble inclusions within the E. coli host cells. The recombinant 60 kDa CrPs were purified and used to raise high titre polyclonal antisera. In immunoblot analysis these antisera reacted with the 60 kDa CrPs from purified elementary bodies of all three chlamydial species in a genus-specific manner. Further molecular analysis allowed the genus-specific cross-reacting epitopes to be localized by using overlapping synthetic peptides covering the C. trachomatis 60 kDa CrP. Immunogold labelling experiments, using purified infectious elementary bodies from the three chlamydial species indicated that the 60 kDa CrPs are not surface accessible to antibody binding.

This article has been cited by other articles:

				S.-J. Jiang, C.-C. Kuo, M. W. Berry, A. W. Lee, and L. A. Campbell Identification and Characterization of Chlamydia pneumoniae-Specific Proteins That Activate Tumor Necrosis Factor Alpha Production in RAW 264.7 Murine Macrophages Infect. Immun., April 1, 2008; 76(4): 1558 - 1564. [Abstract] [Full Text] [PDF]

				S. Fadel and A. Eley Chlamydia trachomatis OmcB protein is a surface-exposed glycosaminoglycan-dependent adhesin J. Med. Microbiol., January 1, 2007; 56(1): 15 - 22. [Abstract] [Full Text] [PDF]

				P. R. Lambden, M. A. Pickett, and I. N. Clarke The effect of penicillin on Chlamydia trachomatis DNA replication. Microbiology, September 1, 2006; 152(Pt 9): 2573 - 2578. [Abstract] [Full Text] [PDF]

				F. O. Eko, Q. He, T. Brown, L. McMillan, G. O. Ifere, G. A. Ananaba, D. Lyn, W. Lubitz, K. L. Kellar, C. M. Black, et al. A Novel Recombinant Multisubunit Vaccine against Chlamydia J. Immunol., September 1, 2004; 173(5): 3375 - 3382. [Abstract] [Full Text] [PDF]

				U. Airaksinen, T. Penttila, E. Wahlstrom, J. M. Vuola, M. Puolakkainen, and M. Sarvas Production of Chlamydia pneumoniae Proteins in Bacillus subtilis and Their Use in Characterizing Immune Responses in the Experimental Infection Model Clin. Vaccine Immunol., May 1, 2003; 10(3): 367 - 375. [Abstract] [Full Text] [PDF]

				M. Klein, A. Kotz, K. Bernardo, and M. Kronke Detection of Chlamydia pneumoniae-Specific Antibodies Binding to the VD2 and VD3 Regions of the Major Outer Membrane Protein J. Clin. Microbiol., May 1, 2003; 41(5): 1957 - 1962. [Abstract] [Full Text] [PDF]

				I. Portig, J. C. Goodall, R. L. Bailey, and J. S. H. Gaston Characterization of the Humoral Immune Response to Chlamydia Outer Membrane Protein 2 in Chlamydial Infection Clin. Vaccine Immunol., January 1, 2003; 10(1): 103 - 107. [Abstract] [Full Text] [PDF]

				A. Ciervo, P. Visca, A. Petrucca, L. M. Biasucci, A. Maseri, and A. Cassone Antibodies to 60-Kilodalton Heat Shock Protein and Outer Membrane Protein 2 of Chlamydia pneumoniae in Patients with Coronary Heart Disease Clin. Vaccine Immunol., January 1, 2002; 9(1): 66 - 74. [Abstract] [Full Text] [PDF]

				P. MYGIND, G. CHRISTIANSEN, K. PERSSON, and S. BIRKELUND Detection of Chlamydia trachomatis-specific antibodies in human sera by recombinant major outer-membrane protein polyantigens J. Med. Microbiol., May 1, 2000; 49(5): 457 - 465. [Abstract] [Full Text] [PDF]

				A. Essig, U. Simnacher, M. Susa, and R. Marre Analysis of the Humoral Immune Response to Chlamydia pneumoniae by Immunoblotting and Immunoprecipitation Clin. Vaccine Immunol., November 1, 1999; 6(6): 819 - 825. [Abstract] [Full Text] [PDF]

				K. Bachmaier, N. Neu, L. M. de la Maza, S. Pal, A. Hessel, and J. M. Penninger Chlamydia Infections and Heart Disease Linked Through Antigenic Mimicry Science, February 26, 1999; 283(5406): 1335 - 1339. [Abstract] [Full Text]

				P. Mygind, G. Christiansen, K. Persson, and S. Birkelund Clin. Vaccine Immunol., May 1, 1998; 5(3): 313 - 318. [Abstract] [Full Text]