Epigenetic marker for the identification of IL17 positive T cells in complex samples

摘要

The present invention relates to a method, in particular an in vitro method for identifying IL-17 expressing T cells in a blood and/or tissue sample derived from a mammal, comprising analysing the methylation status of at least one CpG position in the gene IL-17A, wherein a demethylation of said at least one CpG position in said sample when compared to an analogous position in a non IL-17 blood cell is indicative for a IL-17 positive CD4 positive T cell. The analyses according to the invention can identify IL-17 positive T cells and distinguish them from all other cells in complex samples, such as, for example, other blood cells. The present invention furthermore provides an improved method for quantifying IL-17 positive T cells in complex samples based on a comparison of the IL-17A methylation with a methylation of at least one marker selected from the group of CD3, FOXP3, and/or GAPDH. The method can be performed without a step of purifying and/or enriching cells, preferably in whole blood and/or non-trypsinized tissue.

主权项

1. A kit comprising:
a) a bisulfite reagent, and b) a combination of: a forward primer, a reverse primer, and a probe, the combination selected from:
i) the forward primer comprising SEQ ID NO: 4 or 7, the reverse primer comprising SEQ ID NO. 5 or 8 and the probe comprising SEQ ID NO. 6,ii) the forward primer comprising SEQ ID NO: 9 or 10, the reverse primer comprising SEQ ID NO: 11, 12, 13, or 14, and the probe comprising SEQ ID NO: 15 or 16, andiii) the forward primer comprising SEQ ID NO: 18 or 19, the reverse primer comprising SEQ ID NO: 20 or 21, and the probe comprising SEQ ID NO: 22.