Single Nucleotide Polymorphism Typing Going Spatial: In Situ Padlock Probes Targeting mRNA Variants to Identify Haploidentical Cells within the Tissue Environment

Clinical Chemistry, vol. 72, pp. 163–172

Abstract

BACKGROUND: Despite almost 3 decades of research, the mechanisms underlying the bidirectional trafficking of cells at the maternal-fetal interface that gives rise to microchimerism remain poorly understood. A major barrier to progress has been the lack of suitable detection methods capable of distinguishing maternal from fetal cells within the spatial context of the human placenta. To address this, we developed a novel detection method based on padlock probe technology to differentiate haploidentical cells in placental tissues. METHODS: Padlock probes were designed to target single nucleotide polymorphisms (SNPs) present in messenger RNA transcripts. The assays were first validated in cell lines and subsequently applied to placental tissue to assess its ability to distinguish between maternal and fetal cells. RESULTS: We established a panel of 27 assays targeting 3 human leukocyte antigen-A alleles and 12 biallelic SNPs. The method demonstrated high specificity and sensitivity, detecting minor cell populations at dilutions as low as 1:10 000. Proof of concept was obtained in a decidua basalis specimen, showing the assays' capability to distinguish maternal and fetal cells within placental tissue. CONCLUSIONS: We present a novel, sex-unbiased methodology for the in situ visualization of haploidentical (microchimeric) cells. This approach enables the study of maternal-fetal cellular interactions within their native tissues at the maternal-fetal interface.

Authors 10

1. Emiel Slaats

   Medical University of Graz

   Affiliation as printed

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz , Graz ,

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz, Graz, Austria

2. Katja Sallinger

   Medical University of Graz

   Affiliation as printed

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz , Graz ,

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz, Graz, Austria

3. Julia Schönberger

   Medical University of Graz

   Affiliation as printed

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz , Graz ,

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz, Graz, Austria

4. Bernadette Bramreiter

   Medical University of Graz

   Affiliation as printed

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz , Graz ,

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz, Graz, Austria

5. Jaqueline Anholts Aachen Department of Immunology, Leiden University Medical Centre

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Department of Immunology, Leiden University Medical Centre , Leiden ,

   Department of Immunology, Leiden University Medical Centre, Leiden, the Netherlands

6. Daniel Pitz Jacobsen

   Oslo University Hospital · University of Oslo

   Affiliation as printed

   Division of Gynaecology and Obstetrics, Oslo University Hospital , Oslo ,

   Institute of Clinical Medicine, University of Oslo , Oslo ,

   Division of Gynaecology and Obstetrics, Oslo University Hospital, Oslo, Norway

   Institute of Clinical Medicine, University of Oslo, Oslo, Norway

7. Anne Cathrine Staff

   Oslo University Hospital · University of Oslo

   Affiliation as printed

   Division of Gynaecology and Obstetrics, Oslo University Hospital , Oslo ,

   Institute of Clinical Medicine, University of Oslo , Oslo ,

   Division of Gynaecology and Obstetrics, Oslo University Hospital, Oslo, Norway

   Institute of Clinical Medicine, University of Oslo, Oslo, Norway

8. Amin El Heliebi

   Medical University of Graz

   Affiliation as printed

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz , Graz ,

   Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz, Graz, Austria

9. Michael Eikmans Aachen Department of Immunology, Leiden University Medical Centre

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Department of Immunology, Leiden University Medical Centre , Leiden ,

   Department of Immunology, Leiden University Medical Centre, Leiden, the Netherlands

10. Thomas Kroneis corresponding

    Medical University of Graz

    Affiliation as printed

    Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz , Graz ,

    Gottfried Schatz Research Center, Division of Cell Biology, Histology & Embryology, Medical University of Graz, Graz, Austria