Method of assaying an individual for immune impairment

Patent metadata

Field	Value
Publication number	US-10119959-B2
Application number	US-201113168974-A
Country	US
Kind code	B2
Filing date	Jun 25, 2011
Priority date	Jun 25, 2010
Publication date	Nov 6, 2018
Grant date	Nov 6, 2018

Abstract

Embodiments of the present invention provide diagnostic markers of immunosenescence and methods of identifying individuals with impaired immune function based on a combination of such markers obtained from various analyzes, primarily from blood, testing immune function including the analysis of immune cell subset frequencies, gene expression, cytokine and chemokine levels, and signaling responses to stimulation with cytokines (‘cytokine response’). Particular combinations of markers can predict with high accuracy whether an individual will respond to active vaccination and become protected against recurring diseases.

First claim

What is claimed is:

1. A method of assaying an individual for immune impairment based on an assessment of cytokine responses, the method comprising:
receiving an individual blood sample from an individual of a given age;
separating a baseline sample and a response sample from the individual blood sample;
permeabilizing the baseline sample and the response sample;
stimulating the response sample using at least one cytokine;
staining the baseline sample and the response sample using at least one STAT protein;
measuring the phosphorylation of the at least one STAT protein within the baseline sample and the phosphorylation of the at least one STAT protein within the response sample based on phosphoprotein abundance;
measuring the cytokine stimulation response of the individual by comparing the determined phosphorylation levels of STAT proteins of the baseline sample with the phosphorylation levels of STAT proteins of the response sample;
generating a cytokine response score for the individual based on a comparison of the cytokine stimulation response of the individual to cytokine response profiles from a database of cytokine response profiles;
phenotyping the individual for cytokine response based on the cytokine response score,
wherein an individual is defined as a cytokine responder phenotype if the cytokine response score is within two standard deviations from the average cytokine response score of the cytokine response profiles from the database of cytokine response profiles; and
wherein an individual is defined as a cytokine non-responder phenotype if the cytokine response score is more than two standard deviations from the average cytokine response score of the cytokine response profiles from the database of cytokine response profiles, and wherein the cytokine non-responder phenotype is associated with immune impairment; and
treating the individual based on the phenotype of the individual, wherein the treatment is vaccination for a recurring disease that is appropriate for the cytokine responder phenotype.

2. The method of claim 1 wherein the at least one STAT protein comprises at least one of STAT1 protein in B-cells, STAT1 protein in T-cells, STAT3 protein in B-cells, STAT3 protein in T-cells, STAT5 protein in B-cells, STAT5 protein in T-cells, and STAT1 in monocytes.

3. The method of claim 1 wherein the at least one cytokine comprises at least one of IFN-α, IFN-γ, IL6, IL7, IL10 or IL21.

4. The method of claim 1 , wherein the phenotyping of the peripheral blood mononuclear cell sample is performed using flow cytometry centrifugation.

5. The method of claim 1 wherein the cytokine response score is generated using a Z-score transformation.

6. The method of claim 1 wherein the individual is over 60 years of age.

7. The method of claim 1 wherein the permeabilizing of the baseline sample and the response sample is performed using methanol.

8. The method of claim 1 , wherein comparing the determined phosphorylation levels of STAT proteins of the baseline sample with the phosphorylation levels of STAT proteins of the response sample further comprises calculating a ratio of fold changes between the baseline sample and the response sample.

9. The method of claim 1 , wherein the cytokine response profiles have been previously determined using a regression analysis on measurements of age-associated traits that show a statistically significant change in expression with age.

10. The method of claim 1 , wherein the given age of the individual is at least 60 years old.

Assignees

• Shen Orr Shai S
• Butte Atul J
• Davis Mark M
• Furman David
• Kidd Brian A
• Univ Leland Stanford Junior

Inventors

• Shen-Orr Shai S
• Butte Atul J
• Davis Mark M
• Furman David
• Kidd Brian A

Classifications

• G01N2333/70517

  CD8 · CPC title

• G01N2333/52

  Assays involving cytokines · CPC title

• G01N2333/70514

  CD4 · CPC title

• G01N2800/60

  Complex ways of combining multiple protein biomarkers for diagnosis · CPC title

• G01N33/5091 Primary

  for testing the pathological state of an organism · CPC title