IP Library Granted Patent US 9,746,459
Granted Patent B2
US 9,746,459 · App. 14/052,391 · Granted Aug 29, 2017

Antigen presenting cell assay

Inventors: Rakesh Sindhi (Pittsburgh, PA); Chethan AshokKumar (Pittsburgh, PA)
Assignee: University of Pittsburgh—Of the Commonwealth System of Higher Education
G01N33/5091G01N2800/245
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Quick Facts
Patent No.
US 9,746,459
App. No.
14/052,391
Granted
Aug 29, 2017
Kind
B2
Abstract

Disclosed herein are methods for diagnosing or predicting acute cellular and/or humoral rejection in a subject. In one example, a method of assessing organ rejection includes contacting a first sample comprising antigen presenting cells (APCs) obtained from a subject in need of or having received an organ transplant with a donor antigen from a donor under conditions sufficient to induce uptake of the donor antigen; contacting a second sample comprising APCs obtained from the subject in need of or having received an organ transplant with a third-party antigen under conditions sufficient to induce uptake of the third-party antigen; and determining an antigen presenting index by determining a ratio of uptake of the donor antigen in the first sample to uptake of the third-party antigen in the second sample, wherein the ratio of greater than one indicates organ rejection and the APCs are monocytes or monocyte-derived cells.

Claims (13)

1. A method, comprising:

(i) contacting a first portion of a biological sample comprising antigen presenting cells (APCs) obtained from a subject in need of or having received an organ transplant from a donor, with a donor antigen from the donor in vitro under conditions sufficient to induce uptake of the donor antigen; (ii) contacting a second portion of the biological sample comprising APCs obtained from the subject in need of or having received an organ transplant with a third-party antigen in vitro under conditions sufficient to induce uptake of the third-party antigen; and (iii) measuring amount of uptake of the donor antigen in the first portion of the biological sample and measuring amount of uptake of the third-party antigen in the second portion of the biological sample, wherein the APCS are monocytes or monocyte-derived cells, wherein the method determines the ratio of uptake of the donor antigen in the first sample and the uptake of the third party antigen by the APCs in the second sample.

2. The method of claim 1 , wherein measuring amount of uptake of a donor antigen to uptake of a third-party antigen comprises (i) detecting at least one biomarker following treatment with the donor antigen with the first portion of the biological sample and the third-party antigen with the second portion of the biological sample, wherein the at least one biomarker includes CD14; and (ii) measuring amount of the at least one biomarker CD14 detected following treatment with donor antigen to those detected following treatment with the third-party antigen to determine the antigen presenting index.

3. The method of claim 1 , wherein measuring amount of uptake of donor antigen to uptake of third-party antigen further comprises labeling the donor antigen and third-party antigen with a fluorescent molecule prior to contacting the first portion of the biological sample with the donor antigen or the second portion of the biological sample with the third-party antigen.

4. The method of claim 1 , wherein the method is used to diagnose or predict organ transplant rejection, diagnose or predict graft versus host disease or predict immunity.

5. The method of claim 1 , wherein the method is used to diagnose or predict graft versus host disease after solid organ, bone-marrow, or stem cell transplantation, or a combination thereof.

6. The method of claim 1 , wherein the method is used to predict immunity to an autoantigen, tumor antigen, pathogen antigen, or a combination thereof.

7. The method of claim 1 , wherein monocyte-derived cells are monocytes or monocyte-derived dendritic cells.

8. The method of claim 1 , wherein the subject is a solid organ, bone marrow, stern cell, or combination thereof, recipient or candidate recipient.

9. The method of claim 1 , wherein the subject is a child.

10. The method of claim 1 , wherein the subject has acute cellular and/or humoral rejection.

11. The method of claim 1 , wherein monocyte-derived cells are dendritic cells.

12. The method of claim 1 , wherein the organ is a liver.

Assignments (2)
CONFIRMATORY LICENSE Recorded Dec 17, 2013
From: UNIVERSITY OF PITTSBURGH
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 031832/0079 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 15, 2013
From: SINDHI, RAKESH; ASHOKKUMAR, CHETHAN
To: UNIVERSITY OF PITTSBURGH - OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION
Reel/Frame 031609/0930 →
Continuity (4)
Continuation In Part 13639428
Provisional Application 61322234 · Apr 8, 2010
Provisional Application 61713419 · Oct 12, 2012
Related Publication 20140057315A1 · Feb 27, 2014