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. 2020 Jan 23;51:6. doi: 10.1186/s13567-020-0735-y

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Cell mediated immune responses. A Stimulation index. Mice were immunized with PBS, vector-only strain, and Salmonella secreting Brucella L7/L12. Two weeks post-immunization, 5 mice from each group were sacrificed, and splenocytes were aseptically collected and restimulated with immunizing antigen. Cell proliferative response was measured by MTT based assay after 3 days of incubation. ***Indicates significant difference compared to PBS control; ns: non-significant difference. Significance was determined at P < 0.05. Exact probability is demonstrated on graph. B Stimulation index at 3 weeks. The stimulation index was determined at three weeks of immunization against purified L7/L12 antigen and whole Brucella cell lysate. ***Indicates the significant difference against the PBS control, ns: non-significant difference. C Cytokine response. Mice were immunized with PBS, vector-only strain, and Salmonella secreting Brucella L7/L12. Two weeks post-immunization, 5 mice from each group were sacrificed, and splenocytes were aseptically collected and restimulated with immunizing antigen. After 1 day of incubation, interferon-gamma and IL-4 response were evaluated at mRNA level by quantitative RT-PCR. Fold change of expression is demonstrated. ***Indicates significant difference against PBS control group. Level of significance was determined at P < 0.05. Exact probability value is demonstrated on the graph. D Flow cytometry. Mice were immunized with PBS, vector-only strain, and Salmonella secreting Brucella L7/L12. Two weeks post-immunization, 5 mice from each group were sacrificed, and splenocytes were aseptically collected and restimulated with immunizing antigen. After 1 day of incubation, cells were labeled with CD3+. CD4+ and CD8+ specific anti mouse antibodies CD3+: PE, CD4+: PerCP Vio700, CD8+: FITC (Miltenyi Biotec, Gladbach, Germany) and analyzed in Macsquant flow cytometer instrument (Miltenyi Biotec, Gladbach, Germany). Each population was gated and quantified. ***Indicates significant difference against PBS control group.