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. 2015 Nov 17:3:46.
doi: 10.1186/s40425-015-0087-8. eCollection 2015.

Quantitative real-time PCR assisted cell counting (qPACC) for epigenetic - based immune cell quantification in blood and tissue

Thomas Oliver Kleen  1 , Jianda Yuan  2
Affiliations

Quantitative real-time PCR assisted cell counting (qPACC) for epigenetic - based immune cell quantification in blood and tissue

Thomas Oliver Kleen et al. J Immunother Cancer. .

Abstract

A novel form of immune cell quantification in blood and tissue is described using epigenetic - based, quantitative real-time PCR assisted cell counting (qPACC). The methylation status of the chromatin structure of either actively expressed or silenced genes is the basis of the epigenetic-based cell identification and quantification technology. Discovery of cell type specific removal of methyl groups (demethylation) from the 5'-carbon of the cytosine base in the dinucleotide cytosine phosphate guanine permits precise and robust quantification of immune cells from only small amounts of human blood or tissue samples. These epigenetic biomarkers located on genomic DNA are stably associated with a cell type of interest.

Keywords: Biomarker; Cell quantification; Cell-mediated immunity; Epigenetic; Immune monitoring.

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Figures

Fig. 1
Fig. 1
Assay principle – BSC treated genomic DNA specific qPCR. a Only demethylated cytosines in the target cell type (i.e. CD4+ T-cells) convert to uracil during bisulfite conversion (BSC). This allows BSC genomic DNA specific segregating primer and probes to bind and lead to a qPCR product. b Methylated cytosines in non - target cells do not change during BSC treatment of genomic DNA leading to a primer mismatch and no qPCR product
See this image and copyright information in PMC

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