IP Library › Granted Patent US 12,286,675
Granted Patent B2
US 12,286,675 · App. 16/379,260 · Granted Apr 29, 2025

Epigenetic markers for the identification of blood sub-cells of type 1

Inventors: Sven Olek (Berlin, DE); Ivana Turbachova (Berlin, DE)
Assignee: Precision for Medicine GmbH
C12Q1/6881C12Q2600/154C12Q2600/158
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Quick Facts
Patent No.
US 12,286,675
App. No.
16/379,260
Granted
Apr 29, 2025
Kind
B2
Abstract

The present invention relates to a method, in particular an in vitro method, for identifying CD3CD4 positive T lymphocytes of a mammal, wherein said method comprises analysing the methylation status of at least one CpG position in the CD3a/b/c/d/g genes, in particular their “upstream” regulatory regions, and in particular the promoter and other conserved regions of the gene cd3, wherein a demethylation of at least one CpG in the analyzed sample to at least 90% is indicative for memory and naive CD4 or/and memory and/or naive T lymphocytes. Furthermore, the present invention is directed at the use of DNA-methylation analysis of the genes CD3a/b/c/d for the detection and quality assurance and control of T lymphocytes. Furthermore, the present invention relates to a kit for performing the above methods as well as respective uses thereof. In a preferred embodiment, the present invention furthermore provides an improved method for analysing the methylation status of at least one CpG position in the gene CD3, allowing for a precise analysis even from sub-optimal quality samples, such as non-freshly obtained blood or serum samples.

Claims (9)

1. A method for producing an amplicon from a bisulfite treated human CD3 genetic region, the method comprising:

a) bisulfite treating genomic DNA obtained from a human sample to produce bisulfite treated DNA, and

b) amplifying a CD3 genetic region from the bisulfite treated DNA with at least one primer pair to produce the amplicon,

wherein the amplicon comprises the sequence of SEQ ID NO: 43, SEQ ID NO: 55, or SEQ ID NO: 60.

2. The method according to claim 1 , wherein the amplifying is performed from the 5′ region upstream from the transcription start, and/or promoter region of CD3D gene or CD3G gene.

3. The method according to claim 1 , further comprising detecting methylation status of at least one CpG position by a method selected from methylation specific enzymatic digests, bisulphite sequencing, analysis selected from promoter methylation, CpG island methylation, methylation-specific polymerase chain reaction (MSP), quantitative polymerase chain reaction (qPCR), methylation-sensitive single-nucleotide primer extension (Ms-SNuPE) or other methods relying on a detection of amplified DNA.

4. The method according to claim 1 , wherein said sample is selected from a human body fluid, a human blood sample, a tissue, an organ, a cell type blood sample, or a sample of blood lymphocytes.

5. The method according to claim 1 , wherein said human suffers from autoimmune diseases, transplant rejections, cancer, and/or allergy.

6. The method of claim 1 , wherein the amplicon comprises SEQ ID NO: 55 and SEQ ID NO: 43 or SEQ ID NO: 55 and SEQ ID NO: 60.

Assignments (2)
CHANGE OF NAME Recorded Mar 2, 2021
From: EPIONTIS GMBH
To: PRECISION FOR MEDICINE GMBH
Reel/Frame 056065/0607 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 28, 2020
From: OLEK, SVEN
To: EPIONTIS GMBH
Reel/Frame 051638/0890 →
Priority Claims (1)
EP 08021838 · Dec 16, 2008 · regional
Continuity (3)
Continuation 15477768 · Apr 3, 2017
Continuation In Part 13139808
Related Publication 20190241957A1 · Aug 8, 2019
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