Identification of the new T-cell-stimulating antigens from Mycobacterium tuberculosis culture filtrate

The proteins secreted by Mycobacterium tuberculosis are an important target for vaccine development. To identify the antigens from M. tuberculosis culture filtrate (CF) that strongly stimulate T-cells, the CF was fractionated by ion-exchange chromatography and then non-reducing sodium dodecyl sulfat...

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Bibliographic Details
Published in:FEMS microbiology letters 2004-03, Vol.232 (1), p.51-59
Main Authors: Jae-Hyun, Lim, Hwa-Jung, Kim, Lee, Kil-Soo, Eun-Kyeong Jo, Chang-Hwa, Song, Jung, Saet-Byel, Su-Young, Kim, Lee, Ji-Sook, Tae-Hyun Paik, Jeong-Kyu Park
Format: Article
Language:English
Subjects:
Antigens
Blood
Cell culture
Electrophoresis
Elution
Filtrate
Gel electrophoresis
Interferon
Interleukin 12
Ion exchange
Ion-exchange chromatography
Leukocytes (mononuclear)
Lymphocytes T
Microbiology
Mycobacterium tuberculosis
Peripheral blood mononuclear cells
Polyacrylamide
Proteins
Reactors
Sodium dodecyl sulfate
Sodium lauryl sulfate
Tuberculin
Tuberculosis
Vaccine development
Vaccines
γ-Interferon
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Summary:The proteins secreted by Mycobacterium tuberculosis are an important target for vaccine development. To identify the antigens from M. tuberculosis culture filtrate (CF) that strongly stimulate T-cells, the CF was fractionated by ion-exchange chromatography and then non-reducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis with mini-whole gel elution. Each fraction was screened for its ability to induce interferon-gamma (IFN-γ) production in peripheral blood mononuclear cells isolated from healthy tuberculin reactors. The protein bands that strongly induced IFN-γ production were subjected to N-terminal sequencing. Two new proteins, a 17-kDa protein (Rv0164, MTSP17) and an 11-kDa (Rv3204, MTSP11) protein, were identified. The recombinant MTSP17 (rMTSP17) and rMTSP11 induced significant production of IFN-γ and interleukin (IL)-12p40 in peripheral blood mononuclear cells from healthy tuberculin reactors. Interestingly, IL-12p40 production in response to rMTSP11 was significantly higher than that in response to rMTSP17 or the three components of the antigen 85 complex. These results suggest that MTSP11 antigen should be further evaluated as a component of a subunit vaccine.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-10970400018-7