ABSTRACT
Objective:To investigate the in vitro influence of IL-24 on the functions of CD8 + T cells from patients with non-small cell lung cancer (NSCLC). Methods:Twenty-eight NSCLC patients and 17 healthy individuals were enrolled in this study. Peripheral blood mononuclear cells (PBMC) and bronchoalveolar lavage fluid (BALF) were collected to isolate CD8 + T cells. Real-time reverse transcription-PCR was used to detect the expression of IL-24 receptors (IL-20R1, IL-20R2 and IL-22R1) at mRNA level in CD8 + T cells. Changes in the expression of perforin and granzyme B were measured by flow cytometry after stimulating purified CD8 + T cells with different concentrations of recombinant human IL-24 (10 ng/ml and 100 ng/ml). In vitro direct and indirect contact co-culture systems were established for CD8 + T cells and NSCLC cell line (NCI-H1882 cells). CD8 + T cells induced target cell death and expression of IFN-γ and TNF-α in response to IL-24 stimulation were analyzed. Student′s t test or LSD- t test was used for intergroup comparison. Results:The expression of IL-22R1 at mRNA level was not detected in CD8 + T cells. No significant difference in IL-20R1 or IL-20R2 expression at mRNA level in CD8 + T cells was observed between healthy individuals and NSCLC patients, or between non-tumor sites and tumor sites ( P>0.05). Perforin and granzyme B expression was significantly reduced in CD8 + T cells from peripheral bloods and tumor sites of NSCLC patients as compared with those from healthy individuals and non-tumor sites (all P<0.05). Low concentration of IL-24 (10 ng/ml) did not affect perforin or granzyme B expression in CD8 + T cells ( P>0.05), but high concentration of IL-24 (100 ng/ml) significantly enhanced the expression of perforin and granzyme B in CD8 + T cells from NSCLC patients ( P<0.05). In the direct contact co-culture system, increased ratio of dead target cells and up-regulated IFN-γ and TNF-α expression were induced after stimulating CD8 + T cells from tumor sites in NSCLC patients with high concentration of IL-24 (100 ng/ml), but low concentration of IL-24 (10ng/ml) had no significant influence on CD8 + T cell-induced target cell death and cytokine production. In the indirect contact co-culture system, neither target cell death nor cytokine production induced by CD8 + T cells was affected by IL-24 stimulation. Conclusions:High concentration of IL-24 promoted the in vitro cytolytic function of CD8 + T cells from NSCLC patients, but might not influence the in vivo functions of CD8 + T cells.
ABSTRACT
Objective:To investigate the influence of IL-24 on monocyte activity in patients with non-small cell lung cancer (NSCLC) in vitro. Methods:Twenty-five NSCLC patients and 20 healthy controls were enrolled in the current study. Peripheral blood mononuclear cells (PBMCs) and bronchoalveolar lavage fluid (BALF) samples were collected to isolate monocytes. Expression of IL-22R1, IL-20R1 and IL-20R2 at mRNA level in monocytes was semi-quantified by real-time RT-PCR. Flow cytometry was used to detect the expression of Fas ligand (FasL) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and enzyme linked immunosorbent assay was performed to measure the levels of granzyme A, granzyme B and granzyme H after stimulating monocytes with recombinant human IL-24 (10 ng/ml and 100 ng/ml). The monocytes isolated from peripheral blood of NSCLC patients were co-cultured with A549 cells in vitro. Monocyte-induced target cell death in response to IL-24 stimulation was investigated and changes in the cytotoxicity of monocytes were also assessed after inhibiting granzyme B with Z-AAD-CMK. Student′s t test or LSD- t test was used for comparison. Results:IL-22R1 mRNA was not detected in monocytes. There were no remarkable differences in either IL-20R1 mRNA or IL-20R2 mRNA expression in monocytes between healthy controls and NSCLC patients, or between non-tumor site and tumor site ( P>0.05). FasL and TRAIL expression and granzyme secretion were significantly reduced in monocytes from peripheral blood and tumor sites of NSCLC patients as compared with those from controls ( P<0.05). IL-24 stimulation did not affect the expression of FasL or TRAIL or the secretion of granzyme A or granzyme H ( P>0.05). Low concentration of IL-24 (10 ng/ml) did not affect granzyme B secretion ( P>0.05), while high concentration of IL-24 (100 ng/ml) significantly elevated the secretion of granzyme B by monocytes ( P<0.05). Low concentration of IL-24 (10 ng/ml) did not affect the monocyte-induced target cell death ( P>0.05), while high concentration of IL-24 (100 ng/ml) promoted NSCLC patient-derived monocyte-induced target cell death ( P<0.05). Z-AAD-CMK stimulation suppressed the high concentration of IL-24-mediated elevation of monocyte cytolytic function ( P<0.05). Conclusions:High concentration of IL-24 promoted the cytolytic function of monocytes in NSCLC patients through elevating granzyme B secretion in vitro. However, IL-24 might not influence monocyte function in vivo.
ABSTRACT
Objective To observe the effect of budesonide/doxofylline on patients with persistent asthma. Methods Sixty patients with moderate asthma were randomly divided into treatment group and control group. Patients in treatment group accepted the inhalation budesonide 400μg/d and doxofylline tablet 0.2 bid po but those in control group accepted the inhalation budesonide/formoterol 160/4.5μg bid. FeNO, sputum eosinophils, PEF and ACT scores were compared between two groups. Results PEF and ACT scores in treatment group were lower to those in control group (P0.05). After three months, sputum eosinophils in treatment group were better than that in control group (P0.05). Conclusion There is no significant difference in controlling asthma between budesonide/doxofylline and budesonide/formooterol