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ORIGINAL RESEARCH article
Front. Immunol.
Sec. Immunological Tolerance and Regulation
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1575086
This article is part of the Research TopicMaternal-fetal-placental Immune Interactions: Implications for Pregnancy Outcomes and Long-term HealthView all 18 articles
Manuscript for submission at Frontiers in Immunology Research Topic: Maternal-fetal-placental Immune Interactions
Provisionally accepted
Liseanne J Van 'T Hof*
Johanna M Kapsenberg
Jos Drabbels
Lotte Elisabeth Van Der Meeren
Dave Roelen
Michael Eikmans
Marie-Louise P Van Der Hoorn- Leiden University Medical Center (LUMC), Leiden, Netherlands
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Maternal-fetal HLA compatibility influences pregnancy outcome, including preeclampsia risk. Cervical extravillous trophoblasts (EVT) in early pregnancy provide a non-invasive source for fetal genome acquisition, potentially enabling fetal HLA typing for obstetric risk assessment. This study aimed to achieve fetal HLA typing through EVT isolation using HLA-G-coupled nanoparticle immunomagnetic separation (TRIC) and fluorescence-activated cell sorting (FACS). Method: Cervical samples from 32 pregnant women were collected by cytobrush. Saliva and umbilical cord blood (n=13) served as maternal and fetal HLA genotype controls, respectively. Cervical samples from non-pregnant women, primary cultured EVT, and cryo-sectioned term placentas served as controls for cell phenotype, protein expression, and effect of fixation. FACS and TRIC were applied to isolate EVT from maternal cells, followed by RSSO-PCR for HLA typing. EVT presence pre-and postisolation was determined through HLA-G, β-hCG, and Cytokeratin-7 (CK-7) expression. TRIC was optimized by improving antibody-binding-efficiency, and comparing three (nano)beads types and two magnets. Results: Purity and yield of HLA-G + β-hCG + CK-7 + cells after TRIC failed to match pre-isolation HLA-G + cell counts, despite protocol optimization. FACS revealed a fetal HLA genotype. In contrast, only the maternal HLA genotype was detected in TRIC-isolated cells.EVT counts and maternal cell contamination limit reliable fetal HLA typing from cervical samples. Refining non-invasive EVT isolation techniques may enable fetal HLA typing to be included in risk assessment of pregnancy complications.Trophoblast Retrieval and Isolation from the Cervix (TRIC), Fluorescence-activated cell sorting (FACS) Prenatal testing, Extravillous Trophoblast, Human Leukocyte Antigen (HLA), Preeclampsia.The authors thank Joke Moes in helping to collect samples, Marie van Dijk (Amsterdam UMC) for guidance on protocol adjustments, and Prof. Dr. Jannie Borst for progress guidance and textual feedback on the abstract.
Keywords: TRIC (trophoblast retrieval and isolation from the cervix), FACS (fluorescence-activated cell sorting), Prenatal testing, Extravillous Trophoblast, human leukocyte antigen (HLA), Preeclampsia
Received: 11 Feb 2025; Accepted: 18 Apr 2025.
Copyright: © 2025 Van 'T Hof, Kapsenberg, Drabbels, Van Der Meeren, Roelen, Eikmans and Van Der Hoorn. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Liseanne J Van 'T Hof, Leiden University Medical Center (LUMC), Leiden, Netherlands
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