Cell-type-specific gene expression profile by laser capture microdissection on mirror sections

Immuno-laser capture microdissection (Immuno-LCM) has been used to analyze cell-specific gene expression profiles. However, the usefulness of such a technique is frequently limited by RNA degradation. We, therefore, developed a rapid protocol of LCM on mirror sections, which allows for preserving RN...

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Bibliographic Details
Published in:Journal of immunological methods 2022-06, Vol.505, p.113276-113276, Article 113276
Main Authors: Mazzarella, Giuseppe, Iacomino, Giuseppe, Luca, Pasquale De, Iaquinto, Salvatore, Capuano, Fabiana, Troncone, Riccardo, Rotondi Aufiero, Vera
Format: Article
Language:English
Subjects:
Celiac disease
Cell-specific gene expression profiles
Laser capture microdissection
Mirror sections
γδ + IELs
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Summary:Immuno-laser capture microdissection (Immuno-LCM) has been used to analyze cell-specific gene expression profiles. However, the usefulness of such a technique is frequently limited by RNA degradation. We, therefore, developed a rapid protocol of LCM on mirror sections, which allows for preserving RNA integrity. With such a procedure, we investigated cell-type-specific gene expression of γδ intraepithelial lymphocytes (IELs) in untreated celiac disease (CD). An increase in TGF-β mRNA expression levels was observed in γδ + IELs compared to intestinal enterocytes (IEs), whereas anti-inflammatory IL-10 mRNA production from γδ + IELs was lower compared to IEs. In untreated CD patients, the production of anti-inflammatory cytokines by γδ + IELs is suggestive of a regulatory function, thus playing a critical role in limiting inflammation. This work underscores the importance of LCM on mirror sections as a valuable tool to perform cell-type-specific molecular analysis in tissue.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2022.113276