Figure 5.

C57BL/6 mice were vaccinated with either live BCG or γ-irradiated BCG (irradiated BCG) subcutaneously or intravenously. γ-irradiated BCG was unable to stimulate a reduction in mycobacterial burden compared to live BCG which was able to reduce mycobacterial burden by 1 Log₁₀ (A). N = 5–6 mice per group, *p ≤ 0.05. The reporter RAW-Blue® cell line to assess NF-κB and AP-1 activation was used to assess the ability of either live BCG or γ-irradiated BCG to stimulate NF-kB and AP-1 signaling pathways. The activation of NF-kB and AP-1 signaling pathways was similar, regardless of whether the BCG was viable or non-replicating (B). Cytokine mRNA expression was examined in C57BL/6-derived BMDM, stimulated with BCG for 24 h, using the RT2 profiler™ PCR array for mouse cytokines and chemokines (C). Supernatants were also collected from these cultures and assessed for the production of protein cytokines TNF-α, IL-1β, IL-6, and IL-10 (D). The in vivo data is a representative of multiple iterations with N = 5 mice per group. Data are representative of two in vitro studies.