Safety implication of <i>Salmonella</i> based <i>Brucella</i> vaccine candidate in mice and in vitro human cell culture

다국어 초록 (Multilingual Abstract)

<P><B>Abstract</B></P> <P>An anti-<I>Brucella</I> vaccine candidate comprising rough <I>Salmonella</I> vector delivering <I>Brucella</I> antigens was developed. This system provides a platform for live <I>Brucella</I>-free vaccine development as it can mimic active-intracellular infection of <I>Brucella</I> organism. Exploiting this phenomenon thus provides significant protection at a single dose and also re-assured the safety. To date, no human anti-<I>Brucella</I> vaccines are available, owing to the lack of safe and effective formulation. This study investigated the safety of the vaccine formulation in mice model and in vitro human cell cultures. The experiment was designed to determine the LD50 of the vaccine formulation. The vaccine formulation did not induce any mortality even when mice were administered at 8 × 10<SUP>9</SUP> CFU per oral or per subcutaneous (SC), which was 100-times more than the actual vaccine dose intended for mice model. In contrast, wild-type (WT) <I>Salmonella</I> positive control strain induced 100% mortality at 8 × 10<SUP>7</SUP> CFU per mice via oral or SC routes. Interaction of the vaccine with phagocytic (THP-1 derived macrophage) and non-phagocytic (Caco-2) human cell lines as well as human PBMC was investigated. In in vitro experiments, inflammatory and pyretic cytokines TNF-α, and IL-1β inductions were significantly lower in vaccine group as compared to WT group. Further, apoptosis, nitric oxide synthase and cytotoxicity inductions were comparable and not exacerbated, given that the strain is based on a rough bacterial vector that may have endotoxic lipid-A more readily exposed. These findings corroborated that the vaccine formulation is highly safe in mice model and is relatively mild in the induction of inflammatory cytokines and cellular changes in human cell lines.</P>