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. Author manuscript; available in PMC: 2017 Sep 1.
Published in final edited form as: Infect Genet Evol. 2016 Jun 4;43:289–296. doi: 10.1016/j.meegid.2016.06.008

Figure 1. YopE-OVA fusion protein is secreted by Yersinia, and is processed and presented in vivo to stimulate OVA-specific CD8+ T cell responses.

Figure 1

(A) The indicated Y. pseudotuberculosis strains were grown in Yop secretion conditions (Materials and Methods) and processed for western blotting of whole cell lysates and supernatant proteins, with anti-ovalbumin polyclonal antisera. (B) ksgA− strains expressing YopE18::OVA or YopE138::OVA were delivered to animals via intravenous inoculation and 10 days later, spleens removed, single cell suspensions generated and cells incubated with MHC Class I H2-Kb pentamers specific to OVA residues 257–264 and antibodies against CD8 and CD19. Shown are representative histograms showing pentamer and CD8 labeling of CD8+ CD19− cells. (C) Immunization with L. monocytogeneses ΔactA ΔplcB ActA100::OVA generates OVA-specific CD8+ T cells. 3 × 107 CFU of noted L. monocytogeneses strains were delivered to animals via intravenous inoculation. 7 days later spleens were removed, single cell suspensions generated and cells were incubated with H2-Kb OVA257-264 pentamers and antibodies against CD8 and CD19. Shown are representative scatter plots of 2 separated experiments showing pentamer and CD8 labeling of CD8+ CD19− cells (n=3 mice per experiment).