Intracellular T cell cytokines in patients with B cell chronic lymphocytic leukaemia (B‐CLL)

Abstract

Analysis of cytokine production is a tool to functionally characterise T cells. In this study, spontaneous and polyclonal activation induced cytokine production in T cells were assessed by flow cytometry in patients with B-CLL. Patients with progressive disease had a significantly increased number of T cells spontaneously producing IL-2, IL-4 and GM-CSF as compared to healthy donors and patients with non-progressive CLL, which was not the case for TNF-alpha and IFN-gamma producing T cells. However, no difference in the frequency of T cells producing these cytokines was seen comparing patients with non-progressive disease to control donors. Polyclonal activation of B-CLL T cells in vitro induced an increased proportion of T cells producing these five cytokines in patients as well as in control donors, indicating that T cells in CLL patients might have a relatively well preserved functional capacity. However, the increase in GM-CSF, TNF-alpha and IL-4 producing T cells was more marked in CLL patients than in controls. Furthermore, following activation, a higher frequency of cytokine-producing T cells was noted in patients with progressive disease as compared to those with non-progressive disease. The augmented number of cytokine-producing T cells in CLL may indicate an up-regulated capability of T cells to secrete cytokines, especially in patients with progressive CLL. The increased production of the T cell derived cytokines GM-CSF, TNF-alpha, IL-4 and IL-2 is interesting, as these cytokines have previously been shown to support growth of B-CLL leukaemic cells in vitro and as T cells might specifically recognise the autologous leukaemic B cells in vivo. The findings may suggest a role for T cells in the pathogenesis of B-CLL.

CTLA-4 overexpression in CD19+/CD5+ cells correlates with the level of cell cycle regulators and disease progression in B-CLL patients

“…The criteria for diagnosis and disease progression have been previously described. 5 Direct surface and intracellular immunofluorescence staining were performed for the evaluation of the expressions of all studied proteins and the distribution of cell cycle phases. Cells were then analyzed by flow cytometry using a FACScalibur.…”

IL-4 rescues surface IgM expression in chronic lymphocytic leukemia

“…21 IL-4-expressing T cells are expanded in CLL patients, and a higher proportion of IL-4-expressing T cells is evident in patients with aggressive disease. 22 In proliferation centers, CLL cells are in intimate contact with T cells that express abundant IL-4. 23,24 In vitro, IL-4 promotes CLL cell survival.…”

Cytokines Elicited by HSP60 in Periodontitis with and without Coronary Heart Disease

“…Perhaps the stimulation protocol employed in this study was not sensitive enough to detect IL-4 and IL-5 [39]; as PMA/ionomycin stimulation of IL-4 and IL-10 was insufficient for their intracellular detection, often requiring secondary culture with cytokines followed by re-stimulation with mitogen [40]. However, some studies have shown an induction of intracellular IL-4 with only primary stimulation with PMA/ionomycin [38]. Stimulation of cell cultures with human HSP60 [38,41] resulted in a statistically significant increase in mean fluorescence intensity for intracellular TNF-α in the CP and CHD patients compared with gingivitis group (Table 2, Figure 2), but no significant differences were seen amongst the 4 groups for intracellular IFN-γ, IL-2, IL-4 IL-5 and IL-17.…”

“…This is not surprising as unstimulated cells usually produce little or no measurable cytokines [37] except in patients with progressive malignancies [38]. PMA/ionomycin stimulation of cells resulted in an increase in the mean fluorescence intensity of TH1 cytokines in both controls and patients with CP and CHD ( Table 2).…”

“…However, some studies have shown an induction of intracellular IL-4 with only primary stimulation with PMA/ionomycin [38]. Stimulation of cell cultures with human HSP60 [38,41] resulted in a statistically significant increase in mean fluorescence intensity for intracellular TNF-α in the CP and CHD patients compared with gingivitis group (Table 2, Figure 2), but no significant differences were seen amongst the 4 groups for intracellular IFN-γ, IL-2, IL-4 IL-5 and IL-17. In order to ensure HSP60 and not LPS was eliciting the cytokine response, we denatured HSP60 …”