Analysis of immunogenicity of tuberculosis fusion protein consisting of Ag85B, Mpt64 and HspX antigens expressed in replication and dormancy bacilli

Qing Li, Wen Wen Jiang, Yu Luo, Hong Juan Yu, Nan Nan Song, Bing Xiang Wang, Xin Liu, Ying Zhang, Bing Dong Zhu

Research output: Contribution to journalArticle

Abstract

Objective: To construct protective immunity to Mycobacterium tuberculosis latent infection, a novel fusion protein consisting of HspX, the 190 to 198 peptide of Mpt64 and Ag85B, which were confirmed to be the effective protective antigens mainly expressed in the dormancy and exponential phase of growth, was constructed and its immunogenicity was investigated. Methods: Ag85B and Mpt64 190-198-HspX sequences were amplified by PGR and cloned into plasmids pET-28a. The fusion protein, Ag85B-Mpt64 190-498-HspX (AMH) was expressed in E. coli BL21 and purified with Ni-NTA resins. C57BL/6 mice were immunized three times at 2-week intervals subcutaneously with AMH formulated with the adjuvant composed of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCG-PSN). Humoral and cell-mediated immunity responses were analyzed at five weeks after the last injection. Results: AMH was expressed stably in E. coli and could be purified well by Ni-NTA affinity chromatography. C57BL/6 mice immunized with AMH subunit vaccine generated specific cellular and humoral immunologic response to the stimulation of Ag85B, Mpt64 190-198 and HspX. Conclusion: It suggested that AMH was a promising candidate antigen of tuberculosis subunit vaccine.

Original languageEnglish (US)
Pages (from-to)103-107
Number of pages5
JournalChinese Journal of Microbiology and Immunology
Volume29
Issue number2
DOIs
StatePublished - 2009
Externally publishedYes

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Subunit Vaccines
Inbred C57BL Mouse
Bacillus
Tuberculosis
Tuberculosis Vaccines
Escherichia coli
Antigens
Mycobacterium bovis
Affinity Chromatography
Mycobacterium tuberculosis
Cellular Immunity
Nucleic Acids
Polysaccharides
Immunity
Proteins
Plasmids
Peptides
Injections
Growth
Infection

Keywords

  • Ag85B
  • Fusion protein
  • HspX
  • Mycobacterium tuberculosis
  • Subunit vaccine

ASJC Scopus subject areas

  • Immunology
  • Microbiology
  • Virology

Cite this

Analysis of immunogenicity of tuberculosis fusion protein consisting of Ag85B, Mpt64 and HspX antigens expressed in replication and dormancy bacilli. / Li, Qing; Jiang, Wen Wen; Luo, Yu; Yu, Hong Juan; Song, Nan Nan; Wang, Bing Xiang; Liu, Xin; Zhang, Ying; Zhu, Bing Dong.

In: Chinese Journal of Microbiology and Immunology, Vol. 29, No. 2, 2009, p. 103-107.

Research output: Contribution to journalArticle

Li, Qing ; Jiang, Wen Wen ; Luo, Yu ; Yu, Hong Juan ; Song, Nan Nan ; Wang, Bing Xiang ; Liu, Xin ; Zhang, Ying ; Zhu, Bing Dong. / Analysis of immunogenicity of tuberculosis fusion protein consisting of Ag85B, Mpt64 and HspX antigens expressed in replication and dormancy bacilli. In: Chinese Journal of Microbiology and Immunology. 2009 ; Vol. 29, No. 2. pp. 103-107.
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abstract = "Objective: To construct protective immunity to Mycobacterium tuberculosis latent infection, a novel fusion protein consisting of HspX, the 190 to 198 peptide of Mpt64 and Ag85B, which were confirmed to be the effective protective antigens mainly expressed in the dormancy and exponential phase of growth, was constructed and its immunogenicity was investigated. Methods: Ag85B and Mpt64 190-198-HspX sequences were amplified by PGR and cloned into plasmids pET-28a. The fusion protein, Ag85B-Mpt64 190-498-HspX (AMH) was expressed in E. coli BL21 and purified with Ni-NTA resins. C57BL/6 mice were immunized three times at 2-week intervals subcutaneously with AMH formulated with the adjuvant composed of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCG-PSN). Humoral and cell-mediated immunity responses were analyzed at five weeks after the last injection. Results: AMH was expressed stably in E. coli and could be purified well by Ni-NTA affinity chromatography. C57BL/6 mice immunized with AMH subunit vaccine generated specific cellular and humoral immunologic response to the stimulation of Ag85B, Mpt64 190-198 and HspX. Conclusion: It suggested that AMH was a promising candidate antigen of tuberculosis subunit vaccine.",
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T1 - Analysis of immunogenicity of tuberculosis fusion protein consisting of Ag85B, Mpt64 and HspX antigens expressed in replication and dormancy bacilli

AU - Li, Qing

AU - Jiang, Wen Wen

AU - Luo, Yu

AU - Yu, Hong Juan

AU - Song, Nan Nan

AU - Wang, Bing Xiang

AU - Liu, Xin

AU - Zhang, Ying

AU - Zhu, Bing Dong

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N2 - Objective: To construct protective immunity to Mycobacterium tuberculosis latent infection, a novel fusion protein consisting of HspX, the 190 to 198 peptide of Mpt64 and Ag85B, which were confirmed to be the effective protective antigens mainly expressed in the dormancy and exponential phase of growth, was constructed and its immunogenicity was investigated. Methods: Ag85B and Mpt64 190-198-HspX sequences were amplified by PGR and cloned into plasmids pET-28a. The fusion protein, Ag85B-Mpt64 190-498-HspX (AMH) was expressed in E. coli BL21 and purified with Ni-NTA resins. C57BL/6 mice were immunized three times at 2-week intervals subcutaneously with AMH formulated with the adjuvant composed of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCG-PSN). Humoral and cell-mediated immunity responses were analyzed at five weeks after the last injection. Results: AMH was expressed stably in E. coli and could be purified well by Ni-NTA affinity chromatography. C57BL/6 mice immunized with AMH subunit vaccine generated specific cellular and humoral immunologic response to the stimulation of Ag85B, Mpt64 190-198 and HspX. Conclusion: It suggested that AMH was a promising candidate antigen of tuberculosis subunit vaccine.

AB - Objective: To construct protective immunity to Mycobacterium tuberculosis latent infection, a novel fusion protein consisting of HspX, the 190 to 198 peptide of Mpt64 and Ag85B, which were confirmed to be the effective protective antigens mainly expressed in the dormancy and exponential phase of growth, was constructed and its immunogenicity was investigated. Methods: Ag85B and Mpt64 190-198-HspX sequences were amplified by PGR and cloned into plasmids pET-28a. The fusion protein, Ag85B-Mpt64 190-498-HspX (AMH) was expressed in E. coli BL21 and purified with Ni-NTA resins. C57BL/6 mice were immunized three times at 2-week intervals subcutaneously with AMH formulated with the adjuvant composed of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCG-PSN). Humoral and cell-mediated immunity responses were analyzed at five weeks after the last injection. Results: AMH was expressed stably in E. coli and could be purified well by Ni-NTA affinity chromatography. C57BL/6 mice immunized with AMH subunit vaccine generated specific cellular and humoral immunologic response to the stimulation of Ag85B, Mpt64 190-198 and HspX. Conclusion: It suggested that AMH was a promising candidate antigen of tuberculosis subunit vaccine.

KW - Ag85B

KW - Fusion protein

KW - HspX

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