Christoph Sachsenmaier, Germany

Epimune GmbH Business Development

Presenter Of 1 Presentation

Working Party Diagnostics

MOLECULAR DIAGNOSTIC IMMUNE CELL QUANTIFICATION - A NOVEL APPROACH TO IDENTIFY AND MONITOR PATIENTS WITH IMMUNE CELL DYSREGULATION

Lecture Time
08:55 - 09:10
Room
Gold
Date
21.09.2019, Saturday
Session Time
08:25 - 09:25
Presentation Topic
Diagnostics

Abstract

Background and Aims

Lymphocyte profiling is the mainstay of diagnosing and monitoring patients with primary and secondary immune cell dysregulation. We developed a method for lymphocyte quantification based on epigenetic (i.e. DNA methylation) markers specific for various lymphocyte subsets. The method is suitable for the analysis of dried or frozen blood samples. This allows management of immune compromised patients in areas with limited resources for fresh blood sample logistics. It also opens the perspective of near-patient testing using minimally invasive sampling procedures – e.g. via finger prick.

Methods

We identified genomic regions of lymphocyte-specific de-methylation. Real-time PCR assays were developed to specifically amplify de-methylated regions after bisulfite conversion of genomic DNA. Inclusion of housekeeping genes as well as calibrators and standards allows for absolute and relative quantification of lymphocyte subsets as well as quality control of the test run.

Results

We demonstrate perfect correlation of epigenetic immune cell quantification of T-/B- and NK cells with flow cytometry in whole blood samples of healthy volunteers and patients after hematopoietic stem cell transplantation. We also demonstrate successful application of the epigenetic method to dried blood samples of HIV patients as well as newborns.

Conclusions

Our data demonstrate the feasibility of molecular immune cell quantification using epigenetic markers specific for various lymphocyte subsets. This allows molecular diagnostic applications in settings where fresh blood samples are not available in sufficient quality or quantity for flow cytometry - e.g. monitoring of immune-compromised patients in resource-limited areas, early detection of immune dysregulation in newborns and near-patient testing using point-of care technology.

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