i.Mune™ Treg*


i.Mune™ Treg* is a quantitative test to determine the percentages and absolute** counts of human

TSDR+ regulatory T cells and CD3+ T cells in peripheral whole blood or in blood dried on filter paper.

Allows molecular quantification of1):

  • TSDR+ regulatory T cells (Treg)
  • Overall T cells (CD3+)

TSDR = Treg Specific De-methylated Region


The preferred method for accurate quantification of FOXP3+ Treg4)

FOXP3 TSDR is de-methylated in Treg and fully methylated in stimulated CD4+ T cells, that transienly express the FOXP3 protein (Baron et al., 2007)
FOXP3 TSDR is de-methylated in Treg and fully methylated in stimulated CD4+ T cells, that transienly express the FOXP3 protein (Baron et al., 2007)

Total CD4+ T cells show similar percentage of Treg by FOXP3 flow cytometry (A, left 10.4%) and by TSDR DNA-methylation analysis (B, left 12.7%).
Stimulated CD4+ T cells transiently express the FOXP3 protein (A, right 29.7%) but lack DNA de-methylation of the TSDR (B, right 1.4%). Hence, TSDR de-methylation analysis is more specific for Treg quantification than FOXP3 detection by flow cytometry1)4)


Quantification of Treg can be useful in**:

  • IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X-linked inheritance)3)
  • Other primary immune regulatory disorders4) 
  • Autoimmune diseases5)
  • Allergy and asthma6)
  • Graft-vs-host disease post hematopoietic stem cell transplantation7)
  • Solid organ transplantation, tumor prognosis, immunotherapy, pregnancy8)-10)

*i.Mune™Treg is for research use, only and has not been registered as an IVD product and has not been approved or cleared by a regulatory authority.

 

**The above clinical applications have been established using technologies currently being employed in clinical laboratory routine (e.g. flow cytometry). Epimune has demonstrated equivalence of its epigenetic immune cell quantification method to flow cytometry (in whole blood) and the concordance of epigenetic immune cell quantification between whole blood and dried blood spots1)

Literature

  1. Baron U et al. DNA demethylation in the human FOXP3 locus discriminates regulatory T cells from activated FOXP3(+) conventional T cells. Eur J Immunol. 2007 Sep;37(9):2378-89
  2. Baron U et al. Epigenetic immune cell counting in human blood samples for immunodiagnostics. Sci Transl Med. 2018 Aug 1;10 (452)
  3. Barzaghi F et al. Demethylation analysis of the FOXP3 locus shows quantitative defects of regulatory T cells in IPEX-like syndrome. J Autoimmun. 2012 Feb;38(1):49-58
  4. Cepika AM et al. Tregopathies: Monogenic diseases resulting in regulatory T-cell deficiency. J Allergy Clin Immunol. 2018 Dec;142(6):1679-1695
  5. Blokland SLM, et al. Epigenetically quantified immune cells in salivary glands of Sjögren's syndrome patients: a novel tool that detects robust correlations of T follicular helper cells with immunopathology. Rheumatology (Oxford). 2019 Jul 19. pii: kez268. doi: 10.1093/rheumatology/kez268. [Epub ahead of print] PubMed PMID: 31325310
  6. Singh A et al. Th17/Treg ratio derived using DNA methylation analysis is associated with the late phase asthmatic response. Allergy Asthma Clin Immunol. 2014 Jun 24;10(1)
  7. Peccatori J et al. Sirolimus-based graft-versus-host disease prophylaxis promotes the in vivo expansion of regulatory T cells and permits peripheral blood stem cell transplantation from haploidentical donors. Leukemia. 2015 Feb;29(2):396-405
  8. Pohla H et al. High immune response rates and decreased frequencies of regulatory T cells in metastatic renal cell carcinoma patients after tumor cell vaccination. Mol Med. 2013 Feb 8;18:1499-508
  9. Türbachova I et al. The cellular ratio of immune tolerance (immunoCRIT) is a definite marker for aggressiveness of solid tumors and may explain tumor dissemination patterns. Epigenetics. 2013 Nov;8(11):1226-35
  10. Wieczorek G et al. Quantitative DNA methylation analysis of FOXP3 as a new method for counting regulatory T cells in peripheral blood and solid tissue. Cancer Res. 2009 Jan 15;69(2):599-608
Products