This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Rosenkrands, I.
Right arrow Articles by Andersen, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rosenkrands, I.
Right arrow Articles by Andersen, P.

 Previous Article  |  Next Article 

Infect Immun, June 1998, p. 2728-2735, Vol. 66, No. 6
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification and Characterization of a 29-Kilodalton Protein from Mycobacterium tuberculosis Culture Filtrate Recognized by Mouse Memory Effector Cells

Ida Rosenkrands,1 Peter Birk Rasmussen,1 Markus Carnio,2 Susanne Jacobsen,3 Michael Theisen,4 and Peter Andersen1,*

Department of TB Immunology1 and Department of Clinical Biochemistry,4 Statens Serum Institut, Copenhagen, and Department of Biochemistry and Nutrition, Technical University of Denmark, Lyngby,3 Denmark, and Institute of Microbiology, FML Weihenstephan, Technical University Munich, Munich, Germany2

Received 14 August 1997/Returned for modification 20 October 1997/Accepted 9 February 1998

Culture filtrate proteins from Mycobacterium tuberculosis induce protective immunity in various animal models of tuberculosis. Two molecular mass regions (6 to 10 kDa and 24 to 36 kDa) of short-term culture filtrate are preferentially recognized by Th1 cells in animal models as well as by patients with minimal disease. In the present study, the 24- to 36-kDa region has been studied, and the T-cell reactivity has been mapped in detail. Monoclonal antibodies were generated, and one monoclonal antibody, HYB 71-2, with reactivity against a 29-kDa antigen located in the highly reactive region below the antigen 85 complex was selected. The 29-kDa antigen (CFP29) was purified from M. tuberculosis short-term culture filtrate by thiophilic adsorption chromatography, anion-exchange chromatography, and gel filtration. In its native form, CFP29 forms a polymer with a high molecular mass. CFP29 was mapped in two-dimensional electrophoresis gels as three distinct spots just below the antigen 85 complex component MPT59. CFP29 is present in both culture filtrate and the membrane fraction from M. tuberculosis, suggesting that this antigen is released from the envelope to culture filtrate during growth. Determination of the N-terminal amino acid sequence allowed cloning and sequencing of the cfp29 gene. The nucleotide sequence showed 62% identity to the bacteriocin Linocin from Brevibacterium linens. Purified recombinant histidine-tagged CFP29 and native CFP29 had similar T-cell stimulatory properties, and they both elicited the release of high levels of gamma interferon from mouse memory effector cells isolated during the recall of protective immunity to tuberculosis. Interspecies analysis by immunoblotting and PCR demonstrated that CFP29 is widely distributed in mycobacterial species.

* Corresponding author. Mailing address: Department of TB Immunology, Statens Serum Institut, 5 Artillerivej, DK-2300 Copenhagen S, Denmark. Phone: 45 32 68 34 62. Fax: 45 32 68 30 35. E-mail: tbimm{at}ssi.dk.

Infect Immun, June 1998, p. 2728-2735, Vol. 66, No. 6
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Weldingh, K., Rosenkrands, I., Okkels, L. M., Doherty, T. M., Andersen, P. (2005). Assessing the Serodiagnostic Potential of 35 Mycobacterium tuberculosis Proteins and Identification of Four Novel Serological Antigens. J. Clin. Microbiol. 43: 57-65 [Abstract] [Full Text]  
  • Huard, R. C., Chitale, S., Leung, M., Lazzarini, L. C. O., Zhu, H., Shashkina, E., Laal, S., Conde, M. B., Kritski, A. L., Belisle, J. T., Kreiswirth, B. N., Lapa e Silva, J. R., Ho, J. L. (2003). The Mycobacterium tuberculosis Complex-Restricted Gene cfp32 Encodes an Expressed Protein That Is Detectable in Tuberculosis Patients and Is Positively Correlated with Pulmonary Interleukin-10. Infect. Immun. 71: 6871-6883 [Abstract] [Full Text]  
  • Okkels, L. M., Brock, I., Follmann, F., Agger, E. M., Arend, S. M., Ottenhoff, T. H. M., Oftung, F., Rosenkrands, I., Andersen, P. (2003). PPE Protein (Rv3873) from DNA Segment RD1 of Mycobacterium tuberculosis: Strong Recognition of Both Specific T-Cell Epitopes and Epitopes Conserved within the PPE Family. Infect. Immun. 71: 6116-6123 [Abstract] [Full Text]  
  • Brodin, P., Eiglmeier, K., Marmiesse, M., Billault, A., Garnier, T., Niemann, S., Cole, S. T., Brosch, R. (2002). Bacterial Artificial Chromosome-Based Comparative Genomic Analysis Identifies Mycobacterium microti as a Natural ESAT-6 Deletion Mutant. Infect. Immun. 70: 5568-5578 [Abstract] [Full Text]  
  • Rosenkrands, I., Slayden, R. A., Crawford, J., Aagaard, C., Barry, C. E., I., Andersen, P. (2002). Hypoxic Response of Mycobacterium tuberculosis Studied by Metabolic Labeling and Proteome Analysis of Cellular and Extracellular Proteins. J. Bacteriol. 184: 3485-3491 [Abstract] [Full Text]  
  • Musser, J. M., Amin, A., Ramaswamy, S. (2000). Negligible Genetic Diversity of Mycobacterium tuberculosis Host Immune System Protein Targets: Evidence of Limited Selective Pressure. Genetics 155: 7-16 [Abstract] [Full Text]  
  • Skjot, R. L. V., Oettinger, T., Rosenkrands, I., Ravn, P., Brock, I., Jacobsen, S., Andersen, P. (2000). Comparative Evaluation of Low-Molecular-Mass Proteins from Mycobacterium tuberculosis Identifies Members of the ESAT-6 Family as Immunodominant T-Cell Antigens. Infect. Immun. 68: 214-220 [Abstract] [Full Text]  
  • Rosenkrands, I., Weldingh, K., Ravn, P., Brandt, L., Hojrup, P., Rasmussen, P. B., Coates, A. R., Singh, M., Mascagni, P., Andersen, P. (1999). Differential T-Cell Recognition of Native and Recombinant Mycobacterium tuberculosis GroES. Infect. Immun. 67: 5552-5558 [Abstract] [Full Text]  
  • Weldingh, K., Rosenkrands, I., Jacobsen, S., Rasmussen, P. B., Elhay, M. J., Andersen, P. (1998). Two-Dimensional Electrophoresis for Analysis of Mycobacterium tuberculosis Culture Filtrate and Purification and Characterization of Six Novel Proteins. Infect. Immun. 66: 3492-3500 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»