In vivo ibrutinib treatment does not hamper NLC phenotype and function. (a) NLC were absent from the PB during ibrutinib treatment at the time of blood sampling (left: before ibrutinib treatment, center and right: 15 days and 30 days of ibrutinib treatment, respectively). (Representative results from five independent experiments). (b) Flow cytometry analysis of the expression of CD14, CD163, CD68, CD11b and CD33 by NLC before (day 0, D0) or after ibrutinib treatment of the patient for the times shown (15 days or 1, 3 or 12 months), compared with isotype control (gray) after 15 days of differentiation. (Representative results from five independent experiments). (c) Percentage viability of CLL cells from ibrutinib-treated patients when cultured alone (black line) or with their own NLC (gray line). (Data taken from five independent experiments). (d) Percentage viability of CLL cells from untreated or ibrutinib-treated patients after 12 days of culture either alone or in the presence of NLC from untreated or ibrutinib-treated patients. (Data taken from five independent experiments for ibrutinib-treated patients, seven independent experiments for untreated patients). (*P<0.05 and ***P<0.001).