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Cytokine-producing CD4+ T-cells in the spleens of vaccinated mice after intravenous challenge.

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posted on 2009-05-13, 02:19 authored by Shanmugalakshmi Sadagopal, Miriam Braunstein, Cynthia C. Hager, Jie Wei, Alexandria K. Daniel, Markian R. Bochan, Ian Crozier, Nathaniel E. Smith, Hiriam O. Gates, Louise Barnett, Luc Van Kaer, James O. Price, Timothy S. Blackwell, Spyros A. Kalams, Douglas S. Kernodle

(A) Representative plots show IFN-γ+, TNF-α+ and IFN-γ+TNF-α double-positive cytokine production by splenocytes of mice vaccinated subcutaneously with BCG or 3dBCG at 28 days after intravenous challenge with 2×107 CFU of BCG. Splenocytes were re-stimulated on IFN-γ-treated uninfected bone-marrow derived macrophages and BCG-infected macrophages. Percent values represent frequency of cytokine+ CD4+ T cells for each stimulation condition. (B) Number of IFN-γ+, TNF-α+, IL-2+ and dual cytokine+ CD4+ T cells in spleens 26 to 30 days after intravenous challenge with BCG. Each data point represents the number of BCG-specific cytokine+ T cells in one mouse spleen, calculated from percent values of cytokine+ CD4+ T cells, as in (a), and total number of splenocytes isolated from each mouse. The bars represent median values from 7 to 9 mice in each vaccinated group. Media/- mice are age-matched controls that were vaccinated with media but not challenged, C = challenged. The variances of the median values of the three challenged groups were compared by the Kruskal-Wallis non-parametric test: IFN-γ, P = 0.0031; TNF-α, P = 0.0006; IL-2, P = 0.0165; IFN-γ+TNF-α, P = 0.0007; and IFN-γ+IL-2, P = .0131. The Mann-Whitney P values for two group comparisons are displayed in the figure.

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