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Intracellular cytokine expression by B cells in B-cell chronic lymphocytic leukemia


, : Intracellular cytokine expression by B cells in B-cell chronic lymphocytic leukemia. Blood 98(11 Part 2): 285b, November 16

The main function attributed to B cells is the antibody production involved in humoral immune response. On the other hand activated B cells can serve as antigen presenting cells for CD4+ T lymphocytes, which by secreting cytokines control the process of antibodies production and B cell proliferation. However B cells are not passive recipients of external cytokine signals but can be active producers of cytokines. The profile of B-cell derived cytokines in state of malignant transformation may differ from their normal counterparts. In this study we aimed to compare cytokine production capability of B cells of B-cell chronic lymphocytic leukemia (B-CLL) with B cells of healthy blood donors. We focused on IL-2, IL-4, IFNgamma, since investigations of these cytokines production by B cells have produced conflicting results. Thirty untreated B-CLL patients and 12 healthy individuals were studied. Peripheral blood lymphocytes were isolated and stimulated by PMA and ionomycin in the presence of Brefeldin A. Then combined membrane and intracytoplasmic staining procedures were performed using following monoclonal antibodies: anti-CD19FITC, anti-IL-2PE, anti-IL-4PE, anti-IFNgammaPE. The flow cytometry technique was used to analyze labeled cells. Results were shown as percentage of CD19+ cells and mean fluorescence intensity (MFI). There was statistically significant (p<0.0004) lowering of CD19+/IL4+ cell percentage in B-CLL patients compared to controls (0.99+/-0.93 versus 2.25+/-0.99) along with lowering of MFI (73.63+/-37.89 versus 136.79+/-112.52, p<0.02). We detected statistically significant difference between patients and controls in the percentage of CD19+/IFNgamma+ cells (1.46+/-1.04 versus 2.6+/-1.63, p<0.02), however with no difference in MFI. There was no statistically significant difference between patients and controls in both percentage and MFI of IL-2 expression. IL-4 and IFNgamma are the main cytokines involved in inhibition of malignant cells apoptosis, thus the detected lower percentage of CD19+/IL-4+ and CD19+/IFNgamma+ cells as well as MFI of IL-4 expression in patients than in controls may indicate the faster metabolism of these cytokines by malignant cells. The intracellular cytokine expression by B-CLL cells is not an evidence of these cells capability of producing the cytokines, as they may be internalized by these cells as well. Further investigation is required to assess the IL-4 release by B-lymphocytes, however the detected differences in IL-4 and IFNgamma expression between patients and control group indicate the important role of these cytokines in B-CLL.

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