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US 12,630,856 B2Granted
Identification of host-targeting modulators of bacterial uptake and assay for quantifying intracellular bacteria
AssigneeUniversity of Florida Research Foundation, Inc.
InventorDaniel Czyz
Life of this patent
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Abstract
CFU-less assays of quantifying intracellular bacteria and methods of using the CFU-less assays to identify host-targeting modulators of pathogen uptake or killing by host cells are described. Also described are host-targeting modulators of pathogen uptake by host cells and methods of using the host-targeting modulators to treat infection
Claims13
A method for determining the effect of a compound on uptake of pathogens by host cells comprising:
- (a) labeling the pathogens with a detectable marker;
- (b) incubating a sample containing the host cells with the compound;
- (c) adding the labeled pathogens to the sample and incubating for a period of time sufficient for the labeled pathogen to enter the host cells;
- (d) lysing the host cells and collecting a lysate;
- and
- (e) determining a level of the pathogens internalized by the host cells by measuring the amount of label in the lysate;
- and
- (f) comparing the level of the pathogens internalized by the host cells with a level of pathogen internalized by the host cells in a control sample that did not contain the compound;
- wherein an increase in the level of pathogen internalized by the host cells in the sample relative to the level of pathogen internalized by the host cells in the control sample indicates the compound enhances uptake of the pathogen by the host cells, and a decrease in the level of pathogen internalized by the host cells in the sample relative to the level of pathogen internalized by the host cells in the control sample indicates the compound inhibits uptake of the pathogen by the host cells.
6 more elements
A method for identifying a host-targeting therapeutic comprising:
- (a) labeling pathogens with a detectable marker;
- (b) incubating a sample containing host cells with a compound suspected of being a host-target therapeutic;
- (c) adding the labeled pathogens to the sample and incubating for a period of time sufficient for the labeled pathogens to enter the host cells;
- (d) lysing the host cells and collecting a lysate;
- and
- (e) determining a level of the pathogens internalized by the host cells by measuring the amount of label in the lysate;
- (f) comparing the level of the pathogens internalized by the host cells with a level of pathogen internalized by the host cells in a control sample that did not contain the compound;
- wherein an increase in the level of pathogen internalized by the host cells in the sample relative to the level of pathogen internalized by the host cells in the control sample indicates the compound enhances uptake of the pathogen by the host cells, and a decrease in the level of pathogen internalized by the host cells in the sample relative to the level of pathogen internalized by the host cells in the control sample indicates the compound inhibits uptake of the pathogen by the host cells.
5 more elements
Dependencies are inferred from the claim text.