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 ?? intraepithelial lymphocytes (IELs) in untreated celiac disease (CD). An increase in TGF-? mRNA expression levels was observed in ?? + IELs compared to intestinal enterocytes (IEs), whereas anti-inflammatory IL-10 mRNA production from ?? + IELs was lower compared to IEs. In untreated CD patients, the production of anti-inflammatory cytokines by ?? + 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.
Cell-type-specific gene expression profile by laser capture microdissection on mirror sections
Giuseppe MazzarellaSupervision
;Giuseppe IacominoSupervision
;Riccardo TronconeData Curation
;Vera Rotondi Aufiero
Relatore esterno
2022
Abstract
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 ?? intraepithelial lymphocytes (IELs) in untreated celiac disease (CD). An increase in TGF-? mRNA expression levels was observed in ?? + IELs compared to intestinal enterocytes (IEs), whereas anti-inflammatory IL-10 mRNA production from ?? + IELs was lower compared to IEs. In untreated CD patients, the production of anti-inflammatory cytokines by ?? + 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.File | Dimensione | Formato | |
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