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. 2015 Jan 14;35(2):583–598. doi: 10.1523/JNEUROSCI.2439-14.2015

Figure 3.

Figure 3.

HMGB1 signaling induces early innate and adaptive immune activation. A, Cytokine mRNA expression was analyzed in spleens at the indicated time points after MCAO (n = 7 per group and time point). B, Expression levels of IL-1β and TNF-α were verified on protein level in the serum of control mice (isotype IgG) and anti-HMGB1-treated mice without brain lesion (naive) and 24 h after F-90 min MCAO by ELISA (n = 6 per group). A, B, Data are representative of 2 individual experiments per time point. C, The cytokine-inducing function of HMGB1 was tested in vitro. Interferon-γ (left) and IL-12 (right) concentration was analyzed in supernatants of mixed splenocyte cultures stimulated with disulphide-HMGB1 at the indicated concentration and compared with unstimulated samples (control). Bars represent p < 0.05 between indicated groups. Each data point represents one individual experiment. D, Naive CD3+ T cells were cocultured with untreated (vital) or irradiated APCs and stimulated with HMGB1 at the indicated concentration for 24 h. Interferon-γ concentrations were measured in the supernatant by ELISA (data representative of 5 individual experiments). Vital APCs are required for stimulation of IFN-γ production by T cells.