Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM Beads
In the intestinal epithelium, intraepithelial lymphocytes (IELs) coexist with intestinal epithelial cells (IECs). The IELs have an important role in defending the intestinal tract against pathogens and eliminating tumor cells. Anomalies in the absolute IEL count have been reported in various digesti...
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Bio-protocol LLC
2025-05-01
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| author | Corentin Joulain Stéphanie Bessoles Andrada Chiron Guillaume Sarrabayrouse Salima Hacein-Bey-Abina |
| author_facet | Corentin Joulain Stéphanie Bessoles Andrada Chiron Guillaume Sarrabayrouse Salima Hacein-Bey-Abina |
| author_sort | Corentin Joulain |
| collection | DOAJ |
| description | In the intestinal epithelium, intraepithelial lymphocytes (IELs) coexist with intestinal epithelial cells (IECs). The IELs have an important role in defending the intestinal tract against pathogens and eliminating tumor cells. Anomalies in the absolute IEL count have been reported in various digestive diseases. IELs are typically counted using histologic techniques or under light microscopy after isolation of the epithelium. However, these techniques can introduce bias, which might account for the discrepancies in counts from one study to another. Here, we describe a flow cytometry assay for determining the absolute IEL count and the IEL/IEC ratio. We combined a conventional epithelial isolation method with a BD TruCountTM bead-based absolute counting technique to quantify IELs (CD45+ CD326/EpCAM- CD103+CD3+) and IECs (CD45- CD326/EpCAM+) in a C57BL/6 mouse model. |
| format | Article |
| id | doaj-art-e1c3fa2bb1634c33bb67c7503677285e |
| institution | Kabale University |
| issn | 2331-8325 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Bio-protocol LLC |
| record_format | Article |
| series | Bio-Protocol |
| spelling | doaj-art-e1c3fa2bb1634c33bb67c7503677285e2025-08-20T03:48:46ZengBio-protocol LLCBio-Protocol2331-83252025-05-0115910.21769/BioProtoc.5307Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM BeadsCorentin Joulain0Stéphanie Bessoles1Andrada Chiron2Guillaume Sarrabayrouse3Salima Hacein-Bey-Abina4Université Paris Cité, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, Paris, FranceUniversité Paris Cité, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, Paris, FranceUniversité Paris Cité, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, Paris, FranceClinical Immunology Laboratory, Groupe Hospitalier Universitaire Paris-Saclay, Hôpital Bicêtre, Assistance Publique-Hôpitaux de Paris, Le-Kremlin-Bicêtre, FranceUniversité Paris Cité, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, Paris, FranceUniversité Paris Cité, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, Paris, FranceClinical Immunology Laboratory, Groupe Hospitalier Universitaire Paris-Saclay, Hôpital Bicêtre, Assistance Publique-Hôpitaux de Paris, Le-Kremlin-Bicêtre, FranceIn the intestinal epithelium, intraepithelial lymphocytes (IELs) coexist with intestinal epithelial cells (IECs). The IELs have an important role in defending the intestinal tract against pathogens and eliminating tumor cells. Anomalies in the absolute IEL count have been reported in various digestive diseases. IELs are typically counted using histologic techniques or under light microscopy after isolation of the epithelium. However, these techniques can introduce bias, which might account for the discrepancies in counts from one study to another. Here, we describe a flow cytometry assay for determining the absolute IEL count and the IEL/IEC ratio. We combined a conventional epithelial isolation method with a BD TruCountTM bead-based absolute counting technique to quantify IELs (CD45+ CD326/EpCAM- CD103+CD3+) and IECs (CD45- CD326/EpCAM+) in a C57BL/6 mouse model.https://bio-protocol.org/en/bpdetail?id=5307&type=0 |
| spellingShingle | Corentin Joulain Stéphanie Bessoles Andrada Chiron Guillaume Sarrabayrouse Salima Hacein-Bey-Abina Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM Beads Bio-Protocol |
| title | Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM Beads |
| title_full | Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM Beads |
| title_fullStr | Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM Beads |
| title_full_unstemmed | Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM Beads |
| title_short | Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa Using TruCountTM Beads |
| title_sort | standardized flow cytometry method for absolute counting of intraepithelial lymphocytes in the intestinal mucosa using trucounttm beads |
| url | https://bio-protocol.org/en/bpdetail?id=5307&type=0 |
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