Lenalidomide alters NLCs. (A) Secretion of IL-10 in CLL/NLC cocultures in the presence and absence of lenalidomide. Cell culture supernatants of cocultures setup as described in Figure 1 were used for the quantification of IL-10 (n = 8) by flow cytometry using bead arrays. Mean concentrations of 3 technical replicates per sample are depicted. In addition, the mean value and SEM of all biological replicates within one group are indicated. Paired Student t test was performed for significance analysis (*P = .01). (B) Expression of HLA-DR in NLCs differentiated in the presence and absence of lenalidomide. A total of 1 × 106 CD19-enriched CLL cells were cocultured with 1 × 106 CD14-enriched monocytes from HD in the presence or absence of 10 µM lenalidomide. Cells were cultured in 400 µL complete medium in 24-well plates for 14 days with daily addition of lenalidomide. NLCs were harvested by trypsin digestion and expression of HLA-DR was quantified by flow cytometry. Mean values of 3 technical replicates are depicted for each of the 3 CLL samples. Values of lenalidomide-treated and untreated samples of each patient are connected by lines. Paired Student t test was performed for significance analysis (**P = .009).
