Biomarkers of neutrophil deregulation as diagnostic for gingivitis

Patent metadata

Field	Value
Publication number	US-11573228-B2
Application number	US-201917309892-A
Country	US
Kind code	B2
Filing date	Dec 17, 2019
Priority date	Dec 26, 2018
Publication date	Feb 7, 2023
Grant date	Feb 7, 2023

Abstract

Methods of determining one or more ratios of chemokines in gingival crevicular fluid of an individual selected from the group consisting of: MIF:MIP1a, MIF:CXCL1; MIF:CXCL5; M1F:CXCL8; MIF:CXCL2; and MIF:CXCL6 and methods of identifying an individual as having gingivitis comprising determining one or more ratios of the chemokines in gingival crevicular fluid are disclosed. Methods of treating individuals who are identified as having gingivitis by determining one or more ratios of the chemokines in gingival crevicular fluid are disclosed. Also disclosed are methods of monitoring the treatment individuals who have gingivitis.

First claim

The invention claimed is:

1. A method of determining one or more ratios of chemokines in gingival crevicular fluid of an individual selected from the group consisting of: MIF:MIP1a, MIF:CXCL1; MIF:CXCL5; MIF:CXCL8; MIF:CXCL2; and MIF:CXCL6; the method comprising the steps of:
obtaining a sample of gingival crevicular fluid from the individual;
quantifying Macrophage migration Inhibitory Factor (MIF) in the sample;
quantifying one or more additional chemokines in the sample and calculating one or more MIF to additional chemokine ratios, wherein the one or more chemokines is selected from the group consisting of: Macrophage Inflammatory Protein 1a (MIP1a), C-X-C motif chemokine ligand 1 (CXCL1), C-X-C motif chemokine ligand 5 (CXCL5), Interleukin 8 (CXCL8) C-X-C motif chemokine ligand 2 (CXCL2), and C-X-C motif chemokine ligand 6 (CXCL6), and the one or more ratios of MIF to additional chemokines is selected from the group consisting of: MIF:MIP1a, MIF:CXCL1; MIF:CXCL5; MIF:CXCL8; MIF:CXCL2; and MIF:CXCL6.

2. The method of claim 1 wherein MIP1a is quantified and the ratio MIF:MIP1a is calculated.

3. The method of claim 1 wherein two, three, four, five or six additional chemokines in the sample are quantified and two, three, four, five or six MIF to additional chemokine ratios are calculated.

4. The method of claim 1 wherein the MIF and additional chemokines are quantified using a cytometric bead array, flow cytometry or ELISA spot assay.

5. A method of treating an individual who has gingivitis comprising the steps of:
a) identifying the individual as having gingivitis by
i) obtaining a sample of gingival crevicular fluid from the individual;
ii) quantifying Macrophage migration Inhibitory Factor (MIF) in the sample;
iii) quantifying one or more additional chemokines in the sample, wherein the one or more additional chemokines is selected from the group consisting of: Macrophage Inflammatory Protein 1a (MIP1a), C-X-C motif chemokine ligand 1 (CXCL1), C-X-C motif chemokine ligand 5 (CXCL5), Interleukin 8 (CXCL8), C-X-C motif chemokine ligand 2 (CXCL2) and C-X-C motif chemokine ligand 6 (CXCL6);
iv) calculating one or more calculated ratios selected from the group consisting of: MIF:MIP1a, MIF:CXCL1; MIF:CXCL5; MIF:CXCL8; MIF:CXCL2; and MIF:CXCL6; and
v) comparing the one or more calculated ratios to healthy reference ratios of MIF to the same additional chemokines, wherein said healthy reference ratios exist in an individual not having gingivitis and a calculated ratio greater than the healthy reference ratio indicates the individual has gingivitis;

and
b) applying to the individual's oral cavity an oral care composition comprising one or more ingredients selected from the group consisting of: arginine, zinc phosphate, zinc oxide, zinc citrate, triclosan, chlorhexidine digluconate, thymol, menthol, eucalyptol, methyl salicylate, saline, antibiotics and fluoride.

6. A method of monitoring the treatment an individual who has gingivitis comprising the steps of:
a) determining one or more ratios of chemokines in gingival crevicular fluid of an individual selected from the group consisting of: MIF:MIP1a, MIF:CXCL1; MIF:CXCL5; MIF:CXCL8; MIF:CXCL2; and MIF:CXCL6 by
i) obtaining a sample of gingival crevicular fluid from the individual;
ii) quantifying Macrophage migration Inhibitory Factor (MIF) in the sample;
iii) quantifying one or more additional chemokines in the sample, wherein the one or more additional chemokines is selected from the group consisting of: Macrophage Inflammatory Protein 1a (MIP1a), C-X-C motif chemokine ligand 1 (CXCL1), C-X-C motif chemokine ligand 5 (CXCL5), Interleukin 8 (CXCL8), C-X-C motif chemokine ligand 2 (CXCL2) and C-X-C motif chemokine ligand 6 (CXCL6);
iv) calculating one or more ratios selected from the group consisting of: MIF:MIP1a, MIF:CXCL1; MIF:CXCL5; MIF:CXCL8; MIF:CXCL2; and MIF:CXCL6;

b) treating the individual by applying to the individual's oral cavity an oral care composition comprising one or more ingredients selected from the group consisting of: arginine, zinc phosphate, zinc oxide, zinc citrate, triclosan, chlorhexidine digluconate, thymol, menthol, eucalyptol, methyl salicylate, saline, antibiotics and fluoride; and
c) repeating step a), wherein a decrease in the same chemokine ratios indicates effective treatment.

7. The method of claim 6 wherein the MIF and additional chemokines are quantified using a cytometric bead array, flow cytometry or ELISA spot assay.

8. The method of claim 6 wherein MIP1a is quantified and the ratio MIF:MIPa is calculated.

9. The method of claim 8 wherein the MIF and MIP1a are quantified using a cytometric bead array, flow cytometry or ELISA spot assay.

10. The method of claim 5 wherein the MIF and additional chemokines are quantified using a cytometric bead array, flow cytometry or ELISA spot assay.

11. The method of claim 5 wherein MIP1a is quantified and the ratio MIF:MIPa is calculated.

12. The method of claim 11 wherein the MIF and MIP1a are quantified using a cytometric bead array, flow cytometry or ELISA spot assay.

Assignees

• Colgate Palmolive Co

Inventors

• Zenobia Camille
• Trivedi Harsh Mahendra
• Yang Ying

Classifications

• G01N2333/522

  Alpha-chemokines, e.g. NAP-2, ENA-78, GRO-alpha/MGSA/NAP-3, GRO-beta/MIP-2alpha, GRO-gamma/MIP-2beta, IP-10, GCP-2, MIG, PBSF, PF-4 or KC · CPC title

• G01N33/56955 Primary

  involved in periodontal diseases · CPC title

• G01N33/6863

  Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors · CPC title

• G01N33/5008

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics · CPC title

• G01N2333/523

  Beta-chemokines, e.g. RANTES, I-309/TCA-3, MIP-1alpha, MIP-1beta/ACT-2/LD78/SCIF, MCP-1/MCAF, MCP-2, MCP-3, LDCF-1or LDCF-2 · CPC title