Rheumatology Medical News about Glucocorticoid receptor, SLE, Flow cytometry analysis

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Flow cytometry analysis of glucocorticoid receptor expression and binding in steroid-sensitive and steroid-resistant patients with systemic lupus erythematosus
Arthritis Research & Therapy, 07/16/09

Du J et al. – Data suggest that abnormalities of expression and binding of the glucocorticoid receptor (GR) may be involved in tissue resistance to steroids in SLE patients. Determination of GR expression and binding by flow cytometry (FCM) may be useful in predicting the response to steroid treatment in these pts.

Methods

An evaluation of GRs in T lymphocytes and monocytes of SLE pts to determine the cause of resistance to glucocortcoid therapies
35 pts before treatment and 27 age- and sex-matched normal controls
Disease activity scores were determined before and after treatment and used to divide the pts into steroid-resistant (SR) and -sensitive (SS) groups
GRs in T lymphocytes (CD3+) and monocytes (CD14+) were examined by FCM with GR-mAb and FITC-Dex probes before treatment
PBMCs were isolated for in vitro GCs sensitivity assays
Validity of FCM analysis of intracellular staining for GR was evaluated through comparison with Western blot and radioligand binding assay (RLBA) in U937 and K562 cells in vitro

Results

A significant decrease in GR binding and expression in K562 and U937 cells with 10^-6M dexamethasone (Dex) was found vs those w/o
In addition, a positive correlation was found between FCM and RLBA as well as FCM and WB
Expression and binding of both CD3/GR and CD14/GR in SR pts with SLE, detected by FCM, were all lower than those in SS pts with SLE
Whereas there was no significant difference in SS pts and controls
In vitro corticosteroid sensitivity assay indicated that PHA-stimulated tumour TNF-α, IL-12 and IFN-γ secretion was inhibited by Dex in all controls and SS pts vs in SR group
This confirmed patient classification as SR and SS by disease activity index (SLEDAI) score

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