To analyze whether the expression and modulation of T cell receptor (TCR) signaling is dependent on Casitas B lineage lymphoma b (Cbl-b) in T cells from patients with systemic lupus erythematosus (SLE) upon stimulation with a tolerogenic substance.
Peripheral blood mononuclear cells were obtained from 20 patients with SLE (active disease… (More)
Figure 6. Casitas B lineage lymphoma b (Cbl-b) overexpression rescues T cell receptor (TCR) signaling abnormalities related to anergy resistance in CD4 T cells from patients with systemic lupus erythematosus (SLE). CD4 T cells from SLE patients and healthy controls (HC) (n 10, comprising 3 SLE patients with active, untreated disease, 2 SLE patients in clinical remission, and 5 healthy controls) were transfected with the hyaluronan-tagged Cbl-b-pCEFL vector or empty vector (EV), and the phosphorylation of TCR-associated kinases in response to ionomycin treatment (as compared to that in RPMI medium alone as resting control) and short-term restimulation with soluble anti-CD3 and anti-CD28 antibodies was evaluated by Western blotting. Representative Western blots show increased expression of p-MAPK and decreased expression of p-Akt in the lysates from SLE CD4 T cells transfected with Cbl-b and pretreated with ionomycin.