DCs (2.5 × 104/well) were preincubated at different time points with medium or 3 μM SialoL, as indicated. Then, CD4+ T cells (105/well) were added and cultures were incubated for 72 h in the presence or absence of 1 μg/mL OVA (Imject® OVA; Pierce).
For mixed lymphocyte reaction, total splenocytes from BALB/c mice (2.5 × 105/well) were preincubated for 3 h with medium in the presence or absence of SialoL. Subsequently, splenocytes from OT-II mice (2.5 × 105/well) were added and incubated for 72 h.
Proliferation was assessed by adding 10% (v/v) Alamar Blue® (TREK Diagnostic Systems Inc.) in the last 48 h of incubation. Absorbance was measured at 570 nm and 600 nm as previously described (26 (link)).
For mixed lymphocyte reaction, total splenocytes from BALB/c mice (2.5 × 105/well) were preincubated for 3 h with medium in the presence or absence of SialoL. Subsequently, splenocytes from OT-II mice (2.5 × 105/well) were added and incubated for 72 h.
Proliferation was assessed by adding 10% (v/v) Alamar Blue® (TREK Diagnostic Systems Inc.) in the last 48 h of incubation. Absorbance was measured at 570 nm and 600 nm as previously described (26 (link)).
