Detection of an immune response

• US 9,733,247 B2
• Filed: 01/07/2016
• Issued: 08/15/2017
• Est. Priority Date: 02/25/2011
• Status: Active Grant

• Current Assignee: Benaroya Research Institute
• Original Assignee: Benaroya Research Institute
• Inventors: Kwok, William W., Wambre, Erik
• Primary Examiner(s): Rooney, Nora
• Application Number:
  US14/990,126
• Publication Number:
  US 20160116470A1
• Time in Patent Office: 586 Days
• Field of Search: None
• CPC Class Codes: A61K 39/35 Allergens
• CPC Class Codes: G01N 2333/70514 CD4
• CPC Class Codes: G01N 2333/70596 Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2800/24 Immunology or allergic disorders
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: G01N 33/56977 HLA or MHC typing
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Provided herein are methods of detecting and/or monitoring the presence or severity of an immune disorder in a subject, including detecting a frequency of a Th2a subset of CD4+ T cells in a biological sample of the subject. In some embodiments, the detecting includes: (a) detecting a frequency of CD4+ T cells in a biological sample of said subject; (b) detecting a frequency of a Th2a subset of the CD4+ T cells in the biological sample; and (c) comparing the frequency of the Th2a subset with the frequency of the CD4+ T cells.

Claims

1. A method for detecting the presence of CD4+ T cells that are CRTH2 positive and CD161 positive in a biological sample, comprising:

a) providing the biological sample;

b) staining the biological sample with antibodies, said antibodies comprising anti-CD4, anti-CRTH2 and anti-CD161, to thereby provide a stained sample; and

c) performing flow cytometry on the stained sample,

to thereby detect the presence of CD4+ T cells that are CRTH2 positive and CD161 positive in the biological sample.

2. The method of claim 1, wherein said flow cytometry is fluorescent activated cell sorting (FACS).

3. The method of claim 2, wherein said stained sample comprises at least three fluorochromes.

4. The method of claim 2, wherein said FACS is carried out with at least a 3-color combination.

5. The method of claim 1, wherein said antibodies further comprise anti-CD49d, to detect the presence of said CD4+ T cells that are also CD49d positive in the biological sample.

6. The method of claim 5, wherein said antibodies further comprise anti-CD45RB, to detect the presence of said CD4+ T cells that are also CD45RB negative in the biological sample.

7. The method of claim 6, wherein said antibodies further comprise anti-CD27, to detect the presence of said CD4+ T cells that are also CD27 negative in the sample.

8. The method of claim 1, wherein said antibodies further comprise anti-CD45RO, to detect the presence of said CD4+ T cells that are also CD45RO positive in the sample.

9. The method of claim 1, wherein said biological sample is a human tissue sample.

10. The method of claim 9, wherein said biological sample is a whole blood sample or a peripheral blood mononuclear cell (PBMC) sample.

11. The method of claim 10, wherein said whole blood sample or PBMC sample has a volume of from 0.05 to 10 milliliters.

12. The method of claim 10, wherein said whole blood sample or PBMC sample has a volume of from 0.1 to 5 milliliters.

13. The method of claim 1, wherein said biological sample is collected from a subject afflicted with or at risk of developing an allergic disorder.

14. The method of claim 13, wherein said allergic disorder is seasonal rhinoconjunctivitis, an allergy to pet dander, a food allergy, venom anaphylaxis, or asthma.

15. The method of claim 13, wherein said subject is a human subject.

16. The method of claim 15, wherein said subject is between 0 and 5 years of age, and wherein said allergic disorder is a food allergy.