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    Vaccinia expression of Mycobacterium tuberculosis-secreted proteins: tissue plasminogen activator signal sequence enhances expression and immunogenicity of M. tuberculosis Ag85.

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    Authors
    Malin, Adam S
    Huygen, Kris
    Content, Jean
    Mackett, Mike
    Brandt, Lisa
    Andersen, Peter
    Smith, Steven M
    Dockrell, Hazel M
    Affiliation
    Immunology Unit, Department of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, WC1E 7HT, London, UK. adam.malin@bigfoot.com
    Issue Date
    2000-11
    
    Metadata
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    Abstract
    There is increasing evidence to implicate a role for CD8(+) T cells in protective immunity against tuberculosis. Recombinant vaccinia (rVV) expressing Mycobacterium tuberculosis (MTB) proteins can be used both as tools to dissect CD8(+) T-cell responses and, in attenuated form, as candidate vaccines capable of inducing a balanced CD4(+)/CD8(+) T-cell response. A panel of rVV was constructed to express four immunodominant secreted proteins of MTB: 85A, 85B and 85C and ESAT-6. A parallel group of rVV was constructed to include the heterologous eukaryotic tissue plasminogen activator (tPA) signal sequence to assess if this would enhance expression and immunogenicity. Clear expression was obtained for 85A, 85B and ESAT-6 and the addition of tPA resulted in N-glycosylation and a 4-10-fold increase in expression. Female C57BL/6 mice were immunised using the rVV-Ag85 constructs, and interleukin-2 and gamma-interferon were assayed using a co-culture of immune splenocytes and recall antigen. There was a marked increase in cytokine production in mice immunised with the tPA-containing constructs. We report the first data demonstrating enhanced immunogenicity of rVV using a tPA signal sequence, which has significant implications for future vaccine design.
    Citation
    Vaccinia expression of Mycobacterium tuberculosis-secreted proteins: tissue plasminogen activator signal sequence enhances expression and immunogenicity of M. tuberculosis Ag85. 2000, 2 (14):1677-85 Microbes Infect.
    Journal
    Microbes and Infection
    URI
    http://hdl.handle.net/10541/90043
    DOI
    10.1016/S1286-4579(00)01323-X
    PubMed ID
    11137041
    Type
    Article
    Language
    en
    ISSN
    1286-4579
    ae974a485f413a2113503eed53cd6c53
    10.1016/S1286-4579(00)01323-X
    Scopus Count
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    All Paterson Institute for Cancer Research

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