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.

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