title: Identification of the new T-cell-stimulating antigens from Mycobacterium tuberculosis culture filtrate.
abstract: 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-gamma) production in peripheral blood mononuclear cells isolated from healthy tuberculin reactors. The protein bands that strongly induced IFN-gamma 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-gamma 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.
authors.affiliation: Department of Microbiology, College of Medicine, Chungnam National University, 6 Munhwa-dong, Jung-ku, Daejeon 301-747, South Korea.
authors.forename: Jae-Hyun
authors.initials: JH
authors.lastname: Lim
authors.forename: Hwa-Jung
authors.initials: HJ
authors.lastname: Kim
authors.forename: Kil-Soo
authors.initials: KS
authors.lastname: Lee
authors.forename: Eun-Kyeong
authors.initials: EK
authors.lastname: Jo
authors.forename: Chang-Hwa
authors.initials: CH
authors.lastname: Song
authors.forename: Saet-Byel
authors.initials: SB
authors.lastname: Jung
authors.forename: Su-Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Ji-Sook
authors.initials: JS
authors.lastname: Lee
authors.forename: Tae-Hyun
authors.initials: TH
authors.lastname: Paik
authors.forename: Jeong-Kyu
authors.initials: JK
authors.lastname: Park
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D018664:Interleukin-12 / Q000096:biosynthesis; D053762:Interleukin-12 Subunit p40; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology*; D021122:Protein Subunits / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014374:Tuberculin Test
chemical_list: D000942:Antigens, Bacterial; D053762:Interleukin-12 Subunit p40; D021122:Protein Subunits; D011994:Recombinant Proteins; D018664:Interleukin-12; D007371:Interferon-gamma