@article{mbs:/content/journal/jmm/10.1099/jmm.0.46785-0, author = "Rigden, Rachael C. and Jandhyala, Dakshina M. and Dupont, Chris and Crosbie-Caird, Dianna and Lopez-Villalobos, Nicolas and Maeda, Norihiro and Gicquel, Brigitte and Murray, Alan", title = "Humoral and cellular immune responses in sheep immunized with a 22 kilodalton exported protein of Mycobacterium avium subspecies paratuberculosis", journal= "Journal of Medical Microbiology", year = "2006", volume = "55", number = "12", pages = "1735-1740", doi = "https://doi.org/10.1099/jmm.0.46785-0", url = "https://www.microbiologyresearch.org/content/journal/jmm/10.1099/jmm.0.46785-0", publisher = "Microbiology Society", issn = "1473-5644", type = "Journal Article", keywords = "RT, room temperature", keywords = "HRP, horseradish peroxidase", keywords = "P22, 22 kDa exported MAP protein", keywords = "IFN-γ, interferon-gamma", keywords = "dH2O, deionized water", keywords = "MAP, Mycobacterium avium subspecies paratuberculosis", keywords = "p.i., post-immunization", keywords = "Ab, antibody", keywords = "PPDA, avian purified protein derivative", keywords = "Ag, antigen", keywords = "JD, Johne's disease", keywords = "MTB, Mycobacterium tuberculosis", abstract = "An immunogenic 22 kilodalton exported Mycobacterium avium subspecies paratuberculosis (MAP) lipoprotein (P22) was previously identified, and found to belong to the LppX/LprAFG family of mycobacterial lipoproteins. N-terminal polyhistidine-tagged P22 was produced and purified from Escherichia coli. Antibody recognition of P22, and interferon-gamma (IFN-γ) responses in vitro using blood from a sheep vaccinated with Neoparasec, confirmed its immunogenicity. To evaluate the immunogenicity of P22 in vivo, five sheep were immunized with a single dose containing 0.8 mg recombinant P22 protein in adjuvant. Blood was collected at 4, 13 and 29 weeks post-immunization (p.i.) and tested for anti-P22 antibodies and P22-specific IFN-γ production. P22-specific antibodies were detected by Western blot analysis in all five Neoparasec-immunized sheep at the three time points. Three out of five P22-immunized sheep produced P22-specific antibodies for up to 13 weeks p.i., and two gave a response at 29 weeks p.i. Recombinant P22 was able to stimulate significant IFN-γ production in blood of P22-immunized sheep at 13 and 29 weeks p.i. Recombinant P22 also elicited an IFN-γ response in blood of sheep immunized with Neoparasec.", }