Effect of the sphingosine-1-phosphate receptor antagonist FTY720 on γδT cells in the skin of imiquimod-induced psoriasis-like mouse model / 中华皮肤科杂志

Objective To evaluate therapeutic effect of the sphingosine-1-phosphate receptor antagonist FTY720 on the imiquimod-induced psoriasis-like mouse model,and to explore its molecular mechanism.Methods A total of 8 specific pathogen-free (SPF) female C57BL/6 mice were used in this study,and divided into experimental group (n =3) and control group(n =5):each mouse ear was topically treated with 10 mg/d imiquimod for 7 consecutive days,and the experimental group and control group were intraperitoneally injected with FTY720 and phosphate buffer saline (PBS) respectively on days 2,4 and 6.Ear skin thickness was measured every day.These mice were sacrificed on day 8,and histopathological changes of the ear skin were observed.The ear tissues and draining cervical lymph node cells were obtained.Flow cytometry was performed to analyze changes in the proportion of interleukin (IL)-17+ γδT cells in the mouse skin and draining lymph node tissues between the two above groups,and to determine the expression of sphingosine-1-phosphate receptor 1 (S 1P 1) and cutaneous lymphocyte antigen (CLA) on the surface of the γδT cells.The means of two independent samples were compared by using t test.Results After the 2-day topical application of imiquimod,the mouse ear thickness was significantly lower in the experimental group (0.217 mm± 0.003 mm) than in the control group (0.232 mm ± 0.002 mm,t =4.23,P ＜ 0.01) on day 3,and the significant difference existed till day 8 (all P ＜ 0.01).Histopathological examination of the ear skin revealed that the epidermal thickness was significantly lower in the experimental group (18.62 μm ± 0.19 μm) than in the control group (27.79 μm ± 1.58 μm,t =4.35,P ＜ 0.01).Immunofluorescence staining showed that the degree of neutrophil infiltration in the mouse ear tissue was lower in the experimental group than in the control group.As flow cytometry showed,the proportion of neutrophils was significantly lower in the experimental group (1.57％ ± 0.12％) than in the control group (3.03％ ± 0.33％,t =3.31,P =0.016).In the ear tissues,the experimental group showed significantly decreased proportion of γδT cells or IL-17+ γδT cells (4.88％ ± 0.42％,40.53％ ± 1.76％ respectively) compared with the control group (9.45％ ± 1.22％,56.56％ ± 0.66％ respectively;t =2.75,10.27 respectively,both P ＜ 0.05).In the draining lymph nodes,the experimental group showed significantly increased proportion of γδT cells or IL-17+ γδT cells compared with the control group (t =5.781,4.140 respectively,both P ＜ 0.05),and the fluorescence intensity of S1P1 and CLA in the γδT cells was significantly lower in the experimental group than in the control group (P ＜ 0.05).Conclusion FTY720 can alleviate imiquimod-induced psoriasis-like manifestations in mouse models,likely by down-regulating the expression of S1P1 and CLA in γδT cells,increasing the proportion of γδT cells in the draining lymph nodes followed by the decrease of their proportion in the skin,and decreasing the production of IL-17 in skin tissues.

Influence of extracellular signal-regulated kinase 1/2 pathway inhibitor PD98059 on proliferation and killing function of γδT cells cultured in vitro / 临床肝胆病杂志

ObjectiveTo investigate the effect of extracellular signal-regulated kinase 1/2 (ERK1/2) pathway inhibitor PD98059 on the proliferation and killing function of γδT cells cultured in vitro. MethodsMononuclear cells were separated from peripheral blood in healthy subjects and placed in RPMI 1640 complete medium containing zoledronic acid and interleukin-2 to obtain γδT cells through induction and culture. The ERK1/2 specific inhibitor PD98059 was used to block the ERK1/2 signaling pathway in γδT cells, and the proliferation of γδT cells was measured by cell counting, and the killing function of γδT cells was measured by CCK-8 method. Flow cytometry was used to measure the expression of granzyme B and perforin in γδT cells. The t-test was used for comparison of continuous data between groups. ResultsAfter 10 days of culture, the purity of γδT cells reached 87.94%±2.36%. The number of γδT cells treated with PD98059 was significantly lower than that of control cells ［(6.74±0.36)×105/ml vs (9.42±0.31)×105/ml, t=-12.708, P＜0.001］. Compared with the control cells, those treated with PD98059 had significantly higher positive expression rates of granzyme B and perforin (granzyme B: 48.89%±1.31% vs 41.58%±1.58%, t=7.582, P＜0.001; perforin: 65.92%±3.29% vs 33.49%±2.83%, t=15.478, P＜0001) and a significantly higher killing rate of HepG2 cells (69.28%±4.96% vs 48.34%±3.01%, t=11.201, P＜0.001). ConclusionThe ERK1/2 specific inhibitor PD98059 can inhibit the proliferation of γδT cells, but it can enhance the in vitro killing function of γδT cells.

Interleukin-17 in the Inflammatory Bowel Disease / 한양의대학술지

Inflammatory bowel diseases(IBD), including Crohn's disease and ulcerative colitis, are chronic inflammatory states of the intestinal tract. While the exact mechanisms inducing chronic inflammation are still unclear, it is hypothesized that the inflammation is caused in part by an inappropriate immune response to the intestinal microflora. Although inflammatory diseases are not directly linked to patient survival, symptoms of these diseases significantly decrease quality of life. The incidence rate is higher in western people than eastern people, but the incidence rate of IBD in eastern people, including Korean, is increasing. Recently, it has been reported that IL-17 is an important factor that appears to be involved in IBD induction and progression. This report reviews many recent papers reporting the relationship between IBD and IL-17, which may provide an understanding leading to new means of prevention and treatment for IBD.

Potential founction of γδT lymphocytes for cellular immunotherapies in osteosarcoma / 国际肿瘤学杂志

γδT lymphocytes,a numerically small subset of T lymphocytes,have been shown to directly recognize protein antigens without restriction by polymorphic MHC class I or class Ⅱ molecules and their asso-ciated peptide ligands,secrete cytokines,kill tumor cells and display cytolytie activity against various tumors.In osteosarconla patients,γδT lymphocytes can recognize antigens expressed on the surface of osteosarcoma cells,lyse and kill tumor cells.Combined with converntional chemotherapy and surgery,the new γδT lympho-cytes-based cell immunotherapy of osteosarcoma can increase the survival rate greatly.

Blood ?/? ? cells′ expression,effect and receptor polymorphism in systemic lupus erythematosus / 中华风湿病学杂志

Objective To make clear the expression,role and receptor polymorphism of ?/? T cells in peripheral blood in systemic lupus erythematosus (SLE).Methods Total ?/? T cells,V?1 and V?9/V?2 subsets in peripheral blood of 12 normal controls,12 active and 12 stable SLE patients was examined by using cytometry assay.Then,peripheral blood mononuclear cells (PBMC) were stimulated with ?/? specific antigen.TCR ?/? receptor expression and V?2 chain CDR3 (complementary determination region) length display analysis were investigated further in these patients using SSCP method.Results The number of ?/? T cells were remarkably decreased in active SLE patients,and the ?/? T cells did not show proliferation after antigen stimulation in SLE.V?2 was expressed in all of the SLE patients.CDR3 of V?2 genes was polyclonally displayed.Conclusion This result adds more weight to the protective role of ?/? T cells in SLE.It is postulated that ?/? T cells might have deficient function and act as suppressor T cells.But it appears that there are no specific antigens that stimulate ?/? T cells to proliferate leading to the occurrence of SLE.