The Btk inhibitor ibrutinib impairs the immune response against Mycobacterium tuberculosis of monocyte-derived macrophages from CLL patients and healthy donors

KVIATCOVSKY D; PODAZA E; BEZARES F; GAMBERALE R; COLADO A; ALMEJÚN MB; GENOULA M; COUGOULE C; RISNIK D; SASIAIN MC; GIORNADO, M; BORGE, M; BALBOA L

Mar del Plata

Congreso; Reunión Anual de la Sociedad Argentina de Inmunología; 2016

Sociedad Argentina de Inmunología

Ibrutinib is an oral irreversible inhibitor of the Bruton Tyrosine Kinase (Btk) recently approved for CLL treatment. Last year we described that ibrutinib impairs M1 polarization in macrophages from healthy donor´s. In general, alterations on M1 functions can impact on the resistance against intracellular pathogens, such as M. tuberculosis (Mtb), increasing the occurrence of infectious diseases. Therefore, we aimed to explore the effects of ibrutinib on Mtb-response by macrophages from healthy donors and CLL patients. Macrophages were differentiated by culturing monocytes from healthy donors or CLL patients with M-CSF for 7 days. TNFα, IL-8 and IL-10 secretion were measured by ELISA after macrophages or monocytes stimulation with irradiated Mtb for 24 h in the presence or absence of different doses of ibrutinib. Phagocytosis of Mtb-FITC by macrophages was evaluated by flow cytometry. Bacillary loads were determined by colony-forming units assays. Migration in response to CCL5 was also measured in matrigel chambers. We found that ibrutinib significantly diminish the release of TNF-a, IL-10 and IL-8 by Mtb-stimulated macrophages from healthy donors, being the production of TNF-a severely affected even at low concentrations of the drug, while IL-10 and IL-8 secretion was only affected at 3 µM (n=7, p˂0.05). Additionally, ibrutinib enhances macrophage migration in Matrigel (p˂0.05). Although ibrutinib did not affect Mtb phagocytosis (n=10), it promoted a slight increase in the bacillary loads recovered from macrophages at day 3 and 6 post-infection. Finally we observed that 0.3 µM of ibrutinib also affected Mtb-induced TNF-α production in macrophages (n=6, p