CD4+ T-cell lines used to evaluate a Mycobacterium avium subsp. paratuberculosis (MAP) peptide vaccine

Kari Lybeck, Siri K. Sjurseth, Zainab Al-Touama, Heidi Mikkelsen Melvang, Claus Aagaard, Claus Lundegaard, Gregers Jungersen, Peter Andersen, Ingrid Olsen, Stig Tollefsen

    Research output: Contribution to conferenceConference abstract for conferenceResearchpeer-review

    210 Downloads (Pure)


    The aim of the study was to establish a protocol for generation of MAP-specific T-cell lines and to use these lines for evaluation of a peptide vaccine.
    A protocol for culturing T-cell lines from peripheral blood of goats naturally infected with MAP was established. CD4+ T cells were positively selected using an anti CD4 mAb and Dynabeads. Sorted CD4+ cells were cultivated with purified protein derivative from MAP (PPDj) or E. coli sonicate, IL-2, and IL-15. After two cultivation cycles, T cells were tested for recall responses in a proliferative T-cell assay. T-cell line responses were in average 92 % for PPDj, and -3 % for E. coli sonicate. CD4+ T-cell lines stimulated with PPDj showed a 6 fold increase in IFN- γ production compared to controls. These results indicated that the T-cell lines were MAP-specific.
    The protocol was subsequently used to evaluate MAP-specific peptides as vaccine antigens. T-cell lines were now generated by cultivating CD4+ cells with peptides instead of PPDj. Initially, both healthy and MAP-infected goats were vaccinated with 119 peptides defined by in silico analysis. Cellular responses to the peptides were not detected using standard IFN- γ plasma ELISA. However, testing of T-cell lines from the MAP-infected goats identified peptides that induced strong proliferative responses. The 23 peptides inducing the strongest responses were used in a second vaccination trail with healthy goat kids. Vaccinated kids developed strong IFN-γ and antibody responses, and these MAP-specific peptides show great potential for use in a subunit vaccine.
    Generation of T-cell lines was a valuable tool for selecting MAP vaccine antigens, and the protocol can also be applied for identifying vaccine candidates for other diseases.
    Original languageEnglish
    Publication date2015
    Number of pages1
    Publication statusPublished - 2015
    Event5th European Veterinary Immunology Workshop - Vienna, Austria
    Duration: 2 Sept 20154 Sept 2015
    Conference number: 5


    Conference5th European Veterinary Immunology Workshop
    Internet address


    Dive into the research topics of 'CD4+ T-cell lines used to evaluate a Mycobacterium avium subsp. paratuberculosis (MAP) peptide vaccine'. Together they form a unique fingerprint.

    Cite this