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letter
. 2018 Dec;103(12):e598–e601. doi: 10.3324/haematol.2017.187070

Figure 2.

Figure 2.

IFNγ release from stimulated T cells is impaired by idelalisib in chronic lymphocytic leukemia (CLL). (A) ELISpots for IFNγ release were performed on T samples obtained from 5 CLL patients and 4 healthy donors (HD), stimulated or not with antiCD3 antibody, cytomegalovirus (CMV) or CEF peptides, in the presence or absence of 5 μM idelalisib. In the graph, the spot number obtained with the drug is indicated as mean percentage±Standard Error (SE) of the spot number observed with the correspondent stimulation, defined as 100%. Black squares: HD; white squares: CLL. Since control (CTRL, no treatment/no stimulation) and CTRL+ide (idelalisib treatment/no stimulation) conditions showed the same IFNγ spot number (usually zero) for all CLL and HD T samples, they have not been included in the graph. The increased production of IFNγ induced in T cells by each stimulation is significantly decreased by idelalisib, both for CLL and HD (*P<0.05, Student t-test). (B) IFNγ cytokine secretion assays (CSA) were performed on T samples collected from 5 CLL patients, pre-treated or not with 5 μM idelalisib and stimulated or not with antiCD3 antibody. We acquired approximately 100,000 events inside the “lymphocytes” gate and the analysis identified the IFNγ+ lymphocytes inside the CD3+CD8+ or CD3+CD4+ gates. In the graph, the mean percentages±SE of CD3+CD8+IFNγ+ and CD3+CD4+IFNγ+ lymphocytes are represented with black squares and with gray squares, respectively. Control (CTRL, no treatment/no stimulation) and CTRL+ide (idelalisib treatment/no stimulation) show substantially the same percentages of CD3+CD8+IFNγ+ and CD3+CD4+IFNγ+ lymphocytes (P=n.s., Student t-test). With antiCD3 stimulation, these percentages become respectively 2.3 and 3.1-fold higher in comparison to CTRL (*P<0.05, Student t-test). The presence of idelalisib under stimulation reduces the percentages of CD3+CD8+IFNγ+ and CD3+CD4+IFNγ+ lymphocytes by 1.7 and 2.4 times, respectively, if compared to antiCD3 stimulation alone (*P<0.05, Student t-test). n.s.: not significant.