Figure 3
Influence of exposure to IR fibres on the migration and adherence capacity of innate immune CD11b + cells from CIA-affected rats. (A) Representative flow cytometry dot plots show the frequency of FITC-stained CD11b + cells in lymph nodes draining the site of FITC application (DLN) in FITC painting test (see Material and Methods) in CIA rats housed in cages with IR fibre bedding (+IRF rats) and with wooden shaving bedding (−IRF rats). Of note, rats were transferred to cages with IR fibre bedding five days before immunization. Bar graphs show the expression of mRNA for CCL19 and CCL21 in DLNs from +IRF and −IRF rats, as determined by RT-qPCR. Results are represented as 2−dCt relative to β-actin. (B) Representative flow cytometry dot plots show CD11b expression on splenocytes from +IRF and −IRF rats before (upper dot plots) and after (lower dot plots) passage through nylon wool in the adherence assay (see Material and Methods). Scatter plot indicates the adherence index, i.e. the percentage of adherent cells of CD11b + cells from spleens of +IRF and −IRF rats (calculated as indicated in Material and Methods). (A, B) Flow cytometry profiles were generated using FlowJo Software for Windows, Version 7.8. FlowJo, LLC, Ashland, Oregon, USA (https://www.flowjo.com). Bar graphs (A) and (B) scatter plot were created using GraphPad Prism version 7.00 for Windows, GraphPad Software, La Jolla California, USA (https://www.graphpad.com). Results are expressed as mean ± SEM. n = 10 rats/group. ** p ≤ 0.01.
