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Infect Immun, August 1998, p. 3492-3500, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Two-Dimensional Electrophoresis for Analysis of
Mycobacterium tuberculosis Culture Filtrate and Purification
and Characterization of Six Novel Proteins
Karin
Weldingh,1
Ida
Rosenkrands,1
Susanne
Jacobsen,2
Peter Birk
Rasmussen,1
Martin J.
Elhay,1 and
Peter
Andersen1 *
Department of TB Immunology, Statens Serum
Institut, Copenhagen,1 and
Department of
Biochemistry and Nutrition, Technical University of Denmark,
Lyngby,2 Denmark
Received 26 January 1998/Returned for modification 24 March
1998/Accepted 5 May 1998
Culture filtrate from Mycobacterium tuberculosis
contains molecules which promote high levels of protective immunity in
animal models of subunit vaccination against tuberculosis. We have used two-dimensional electrophoresis for analysis and purification of six
novel M. tuberculosis culture filtrate proteins (CFPs): CFP17, CFP20, CFP21, CFP22, CFP25, and CFP28. The proteins were tested
for recognition by M. tuberculosis-reactive memory cells from different strains of inbred mice and for their capacity to induce
a skin test response in M. tuberculosis-infected guinea pigs. CFP17, CFP20, CFP21 and CFP25 induced both a high gamma interferon release and a strong delayed-type hypersensitivity response,
and CFP21 was broadly recognized by different strains of inbred mice.
N-terminal sequences were obtained for the six proteins, and the
corresponding genes were identified in the Sanger M. tuberculosis genome database. In parallel we established a two-dimensional electrophoresis reference map of short-term culture filtrate components and mapped novel proteins as well as already-known CFP.
*
Corresponding author. Mailing address: Department of TB
Immunology, Statens Serum Institut, Artillerivej 5, 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, August 1998, p. 3492-3500, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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