# US 2014/0194321 A1 · Cell-based method to detect skin sensitizers > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0194321 A1 - Document ID: US20140194321A1 - Status: Published application - Kind: A1 - Application number: 13/734,751 - Filed: 2013-01-04 - Published: 2014-07-10 - Earliest claimed: 2013-01-04 - Assignees: Cellomics, Inc. - Applicants: Cellomics, Inc. - Inventors: Monica Jo Tomaszewski - Web page: https://patentgrep.com/patent/US20140194321A1 ## CPC - G01N 33/5047: {Cells of the immune system} - G01N 33/5029: {on cell motility} - G16B 99/00: Subject matter not provided for in other groups of this subclass - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - G16B: BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY ## Abstract A method of predicting whether a compound is a skin sensitizer using an in vitro approach. The method includes a step of imaging immune effector cells positioned within a plurality of containers to obtain imaged cellular targets, each container being treated with a different concentration of a compound. The method further includes a step of quantitatively measuring the imaged cellular targets over a range of concentrations of the compound to detect changes in multiple cellular targets of the immune effector cells associated with skin sensitivity. The method further includes a step of analyzing measurements obtained from the measured imaged cellular targets over the range of concentrations of the compound to determine whether the compound is a skin sensitizer. ## Claims ### Claim 1 A method of predicting whether a compound is a skin sensitizer, the method comprising: imaging immune effector cells positioned within a plurality of containers to obtain imaged cellular targets, each container being treated with a different concentration of a compound; quantitatively measuring the imaged cellular targets over a range of concentrations of the compound to detect changes in multiple cellular targets of the immune effector cells associated with skin sensitivity; and analyzing measurements obtained from the measured imaged cellular targets over the range of concentrations of the compound to determine whether the compound is a skin sensitizer. ### Claim 2 (depends on 1) The method recited in claim 1, wherein the imaging of the immune effector cells is performed using a quantitative high-content cell imaging system. ### Claim 3 (depends on 1) The method recited in claim 1, wherein imaging immune effector cells comprises: using a phenotypic approach on a first portion of the treated immune effector cells; and using a chemotactic approach on a second portion of the treated immune effector cells. ### Claim 4 (depends on 1) The method recited in claim 1, wherein analyzing measurements obtained from the measured imaged cellular targets over the range of concentrations to determine whether the compound is a skin sensitizer comprises: determining data normalization values for each cellular target; determining a representative value for each cellular target at each concentration of the compound; generating a dose response curve for each cellular target based on the representative values of the cellular target; qualifying a dose response of each cellular target based on the dose response curve of the cellular target to determine a reaction of the cellular target to increasing concentrations of the compound; and applying a decision matrix to the qualified dose responses. ### Claim 5 (depends on 4) The method recited in claim 4, wherein determining data normalization values for each cellular target comprises subtracting a baseline response of vehicle-treated cells from measurements of the measured imaged cellular targets over the range of concentrations of the compound. ### Claim 6 (depends on 4) The method recited in claim 4, wherein determining the representative value for each cellular target at each concentration of the compound comprises determining an average value for each cellular target at each concentration of the compound. ### Claim 7 (depends on 4) The method recited in claim 4, wherein generating a dose response curve for each cellular target comprises: plotting the representative value of the cellular target corresponding to each concentration of the compound; and fitting a curve to the plotted values. ### Claim 8 (depends on 4) The method recited in claim 4, wherein qualifying the dose response of each cellular target comprises characterizing the dose response curve of the cellular target as one of: increasing, decreasing, and flat. ### Claim 9 (depends on 8) The method recited in claim 8, wherein characterizing the dose response curve comprises: characterizing the dose response curve as flat if all of the dose response curve is positioned within a predefined range; otherwise: characterizing the dose response curve as decreasing if a highest value of the dose response curve occurs at a lowest concentration of the compound; or characterizing the dose response curve as increasing if a lowest value of the dose response curve occurs at a lowest concentration of the compound. ### Claim 10 (depends on 4) The method recited in claim 4, wherein applying the decision matrix comprises determining whether the compound is a skin sensitizer based upon expected behavior of the qualified dose responses of two or more cellular targets. ### Claim 11 (depends on 4) The method recited in claim 4, wherein applying the decision matrix comprises predicting that the compound is one of: a skin sensitizer, a skin non-sensitizer, or toxic to the skin. ### Claim 12 (depends on 1) The method recited in claim 1, wherein the imaged cellular targets comprise one or more of: a gross marker, a specific marker, and a functional outcome. ### Claim 13 (depends on 1) The method recited in claim 1, wherein the imaged cellular targets includes at least one of cell trafficking and cell motility imaged using a chemotactic approach. ### Claim 14 (canceled) ### Claim 15 A method of screening a plurality of compounds for skin sensitivity, the method comprising: for each compound: imaging immune effector cells positioned within a plurality of containers to obtain imaged cellular targets, each container being treated with a different concentration of the compound, the imaging being performed using a quantitative high-content cell imaging system; quantitatively measuring the imaged cellular targets over a range of concentrations of the compound to detect changes in multiple cellular targets of the immune effector cells associated with skin sensitivity; analyzing measurements obtained from the measured imaged cellular targets over the range of concentrations of the compound to determine if the compound is predicted to be a non-skin sensitizer; and recommending that the compound be further tested if the compound is not predicted to be a non-skin sensitizer. ### Claim 16 (depends on 15) The method recited in claim 15, further comprising performing further testing for skin sensitivity on the compounds that are not predicted to be non-skin sensitizers. ### Claim at position 17 17-18. (canceled) ### Claim 19 (depends on 1) The method recited in claim 1, wherein the immune effector cells comprise U937 cells. ### Claim 20 (depends on 1) The method recited in claim 1, wherein analyzing measurements obtained from the measured imaged cellular targets over the range of concentrations of the compound is also performed to determine whether the compound is toxic to the skin. ### Claim 21 A method of predicting whether a compound is a skin sensitizer, the method comprising: culturing immune effector cells positioned within a plurality of containers; treating each container with a different concentration of a compound; staining the cells in the containers with fluorescent materials; transferring the stained cells to a chemotaxis plate; applying chemokine to the stained cells in the chemotaxis plate; and acquiring images from the stained cells after the chemokine has been applied thereto; quantitatively measuring the imaged cellular targets over a range of concentrations of the compound to detect changes in multiple cellular targets of the immune effector cells associated with skin sensitivity; and analyzing measurements obtained from the measured imaged cellular targets over the range of concentrations of the compound to determine whether the compound is a skin sensitizer. ### Claim 22 (depends on 21) The method recited in claim 21, wherein analyzing measurements obtained from the measured imaged cellular targets comprises: determining data normalization values for each cellular target; determining a representative value for each cellular target at each concentration of the compound; generating a dose response curve for each cellular target based on the representative values of the cellular target; qualifying a dose response of each cellular target based on the dose response curve of the cellular target to determine a reaction of the cellular target to increasing concentrations of the compound; and applying a decision matrix to the qualified dose responses. ### Claim 23 (depends on 22) The method recited in claim 22, wherein generating the dose response curve for each cellular target comprises: plotting the representative value of the cellular target corresponding to each concentration of the compound; and fitting a curve to the plotted values.