SLPI UPREGULATES THE EXPRESSION OF IL-6 AND IL-10 IN LYMPHOCYTES

GUERRIERI DIEGO; MACARENA REITERI; MAFFIA PAULO; CHULUYAN EDUARDO

Córdoba, Argentina

Congreso; VII Latinamerican Congress of Immunology; 2005

ALAI

.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.    Previous works in our laboratory have demonstrated that Secretory Leucocyte Protease Inhibitor (SLPI) decreased the proliferation of human T cells. In this study, we analyzed the expression of different cytokines produced by lymphocytes treated with rhSLPI.  In order to evaluate the capacity of SLPI to modulate the pattern of expression of cytokines we use a BDTM CBA Human Th1/Th2 Cytokine Kit II. This kit allows us to measure simultaneously IL-2, IL-4, IL-6, IL-10, TNF and IFN by flow cytometry in the supernatant of lymphocytes stimulated with rhSLPI (4 mg/ml, 5 days, 37ºC). The presence of rhSLPI increase the expression of IL-6 (from 0,065±0,01 to 3,4±0,7 ng/ml) and IL-10 (from 0,02±0,01 to 0,4±0,1 ng/ml) meanwhile no significant effect was observed on the others cytokines. We also observed that IL-2 (8ng/ml) reverted the increase produced by rhSLPI of IL-6 (1,4±0.7 ng/ml) and IL-10 (0,3±0,04 ng/ml). Furthermore, rhSLPI (4 mg/ml) decreased the expression of T-box expressed in T-cell (T-bet) in lymphocytes stimulated with IL-2 (8 ng/ml) during 4 days through a Western blot. T-bet expression is implicated in generating Th1 type of immune response. These results suggest that SLPI can modulate the expression of cytokines leading to Th2 cytokine pattern of expression.