Ex facilitate maximal bacterial killing, we will assess

Ex vivo infection assay.  Maintaining the
hypothesis that a combination of antibodies with diverse neutralizing
mechanisms and Fc-mediated functions will cooperate to facilitate maximal
bacterial killing, we will assess mAb combinations in the presence of
neutrophils plus complement. Clinical isolates of S. aureus obtained from the Thomsen Lab cohort of invasively
infected children will be incubated with neutrophils at a multiplicity of
infection (MOI) of 10, diluted guinea pig complement, and mAb combinations at
selected doses. CFU and neutrophil viability (via LDH release assay) will be
assessed after 90 minutes of infection. The use of active and heat-inactivated
complement will allow the determination of the complement-mediated contribution
to phagocytosis. The most potent mAb combinations for CFU reduction and
neutrophil protection will provide insight into the critical components of the
host-leukocidin interaction.

 

Selected
combinations from the above experiments will then be assessed in future studies
in the Thomsen Lab, including a whole blood killing assay to further evaluate
the hypothesis that a combination of antibodies with distinct neutralizing
mechanisms and Fc-mediated, immune enhancing functions will cooperate to
facilitate maximal bacterial killing within human blood.  Together, these experiments will test an
important hypothesis and provide insights into host-pathogen interactions.  The most potent mAb combinations identified
in these studies will represent a potential modality for intervention against S. aureus infection in humans, while
also (by assessment of the distinct properties of that combination) providing
novel insights into the critical components of the host-toxin interaction.

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