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Unravelling the mechanism of action of Tofacitinib on immune cell trafficking through IBD-associated microvasculature

ABSTRACT: In order to investigate the effect of tofacitinib on leukocyte trafficking through primary HIMEC from patients with UC, we designed a pilot experiment with the aim to establish the most efficient JAK inhibiting dose. For this purpose, we performed a dose-dependent experiment on UC-derived HIMEC, assessing Peripheral Blood Mononuclear Cells (PBMC) transmigration and using a range between 10 and 300 nM (10, 50, 100, 300 nM), according to the literature. More in detail, in a Transwell system (Figure 1), 500.000 PBMC, in 3 replicates, have been plated on confluent monolayers of untreated or Tofacitinib pre-treated (24 hours) UC-HIMEC at the various concentrations. After 18 hours of incubation transmigrated PBMC have been counted and their phenotype characterized by FACS. In order to avoid any direct effect of Tofacitinib on PBMC, the 18 hour- long incubation has been performed in the absence of the drug.

ORGANISM(S): Homo sapiens

PROVIDER: GSE165636 | GEO | 2021/12/09

REPOSITORIES: GEO

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