0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

RF07.39.1 – Genetic risk factors and peripheral biological markers of conversion from Mild…
c) Flow cytometry analysis of PBMC
In order to accurately compare the soluble factor production by PBMC of
the subject studied, possible differences in the com-position of PBMC populations
should be assessed. Thus, a flow cytometry analysis will be performed
on unfractionated PBMC obtained from the studied subjects. The following
monoclonal antibodies (mAb, all from BD PharMingen, San Diego, CA) will
be used for flow-activated cell sorting (FACS) analysis: anti-HLA-DR, anti-
HLA-ABC, anti-CD80, anti-CD86, an-ti-CD83, anti-CD14, anti-CD1a, anti-
CD11c, anti-CD40, anti-CD3, anti-CD4, anti-CD8, anti-CD154, anti-CD25. Negative
controls will be isotype-matched antibodies. To determine surface cell
phenotype cells will be washed in assay buffer (PBS, 0.5% BSA and 0.1%
sodium azide), incubated with mAbs for 15 minutes at +4°C, washed and then
analyzed by flow cytometry. Analyses will be performed with a FACSCalibur
flow cytometer. Data will be analyzed using CellQuest software.
2009 707