ABSTRACT
To observe the contribution of FoxO1 on its downstream molecules expression and function in Jurkat cells,Jurkat cells were infected with FoxO1 expression lentivirus and interference lentivirus to establish FoxO1-KLF2-S1P1 signaling pathways mod-el.After infection of 120 h,FoxO1,KLF2,S1P1 and CD62L mRNA levels and the protein expression of FoxO1,FoxO1-p and KLF2 were increased (P<0.05) in FoxO1-overexpression group.Converse results(P<0.05) were observed in the interference group.The proportions of S1P1+cells were increased in both groups.It was notably that S1P1+cells were decreased(P<0.05) in interference group after infection of 72 h.The proportion of CD62L+cells was increased(P<0.05) in overexpression group,it was decreased(P<0.05) in the interference group.This vitro experiments showed S1P1 and CD62L could be regulated by FoxO1 lentivirus,and provided an experimental basis for research about intracellular FoxO1-KLF2-S1P1 signaling pathways and cellular functions.
ABSTRACT
Objective To understand how Vibrio vulnificus hemolysin (VvhA) affects the viability of murine liver CD4+ T cells as well as its effects on the numbers of mitochondria and the expression of CD62L. Methods The primary murine liver monocytes (MNs) were isolated from C57BL/ 6 mice and then treated with recombinant VvhA (rVvhA) for 6 hours in vitro. The viability of murine liver CD4+T cells and the expression of CD62L were measured by staining with anti-mouse CD4, CD8, CD44, CD62L and cell via-bility fluorescent dye or fluorescent antibody. Moreover, the cells were simply incubated with MitoTracker or JC-1 probes to label mitochondria and mitochondrial membrane potential, which were further analyzed by using flow cytometry analysis. Results With the increase in the doses of rVvhA, the viability of murine liv-er CD4+T cells was decreased from 81. 5% to 15. 8% . The expression of CD62L on the surface of murine liver CD4+T cells was dramatically decreased. Both the murine liver na?ve and effector CD4+ T cells were sensitive to the cytotoxicity of rVvhA. Moreover, treating murine liver CD4+ T cells with rVvhA resulted in significantly decreased numbers of mitochondria and lower mitochondrial membrane potential. Conclusion The cytotoxicity of rVvhA to murine liver CD4+T cells might be achieved through inhibiting the expression of CD62L, decreasing the numbers of mitochondria and lowering mitochondrial membrane potential.
ABSTRACT
BACKGROUND: The CD4+ T-helper cells comprise functionally distinct subsets of Th1 and Th2 cells that are distinguished on the basis of differential cytokines production. Th1 cells secrete interferon-gamma, lymphotoxin, interleukin-2. Th2 cells produce interleukin-4, interleukin-5, interleukin-10. A previous study shown that Th2 cells and their cytokines increased in patients with atopic asthma. We compared cytokines(IL-4, IFN-gamma) activity and subpopulation of T-lymphocytes in peripheral blood from atopic asthmatics versus non-asthmatics. METHOD: Fifteen patients with atopic asthma(nine men, six women), twelve patients with chronic bronchitis (six men, six women), five healthy persons(three men, two women) were studied. Activity of IL-4, IFN-gamma and T-cell subpopulation in peripheral blood were estimated. RESULTS: Patients had a median age of 55yr. The mean activity of IL-4 of asthmatics was significantly increased(control 0.75+/-1.1pmol/L, atopic asthmatics 3.50+/-0.75pmol/L, chronic bronchitis 2.01+/-1.2pmol/L), but IFN- was not significantly increased. In the T lymphocyte subsets the percent of CD62L+ T-lymphocytes of asthmatics was not significantly increased(control 16.7+/-16.4 %, atopic asthmatics 24.8+/-23.6 %, chronic bronchitis 17.0+/-16.9%). CONCLUSION: In this study elevated production of IL-4 was observed in atopic asthmatics. CD62L+ T-lymphocytes was not increased in atopic asthma.
Subject(s)
Humans , Male , Asthma , Bronchitis, Chronic , Cytokines , Interferon-gamma , Interleukin-10 , Interleukin-2 , Interleukin-4 , Interleukin-5 , Lymphotoxin-alpha , T-Lymphocyte Subsets , T-Lymphocytes , Th1 Cells , Th2 CellsABSTRACT
Objective:To elucidate the characterization of antigen-specific memory T cells from PPD~+ individuals after stimulation with BCG in vitro.Methods:PBMCs were isolated from PPD~ -/+ normal human peripheral blood and stimulated with BCG. The level of IFN-? in the culture supernatants was assayed by ELISA and the frequency of IFN-?-producinging cells was detected by ELISPOT. The subsets and frequency of cytokine-producing cells were determined at a single cell level by flow cytometry.Results:After stimulation with BCG, PBMCs from PPD~+ but not PPD~- individuals produced significantly high levels of IFN-? in culture supernatants detected by ELISA(P