Mechanism of human placenta-derived mesenchymal stem cells in inhibiting TNF-α secretion in Th1 cells through CD73/Nrf2 pathway to reduce liver injury in mice with graft versus-host disease / 中华微生物学和免疫学杂志

Objective:To investigate the mechanism of human placenta derived mesenchymal stem cells (hPMSCs) in the inhibition of TNF-α secretion in CD4 + IFN-γ + T cells (Th1) through CD73/nuclear factor-erythroid 2-related factor 2(Nrf2) pathway to reduce liver injury in mice with graft versus-host disease (GVHD). Methods:Flow cytometry (FCM) was used to analyze the expression of TNF-α in Th1 cells and the expression of PD-1 on CD4 + IFN-γ + TNF-α + T cells (TNF-α + Th1 cells) isolated from peripheral blood and liver tissues of mice with GVHD. Hematoxylin-hosin (HE) staining, Masson staining and immunofluorescence staining were used to observe the pathological changes in liver tissues of GVHD mice in each group. HE staining was also used to observe the pathological changes in skin and lung tissues of GVHD mice. A nonconditional protocol to induce the differentiation of peripheral blood mononuclear cells (PBMCs) into Th1 cells in vitro was established. The proportion of TNF-α + Th1 cells and the mean fluorescence intensity (MFI) of Nrf2 and phosphorylated nuclear factor-kappa B (p-NF-κB) in this T cell subgroup were detected. Results:Compared with the normal control group, the proportion of TNF-α + Th1 cells and the expression of PD-1 on this T cells in peripheral blood and liver tissues of mice in the GVHD high group increased significantly ( P<0.01). The proportion of TNF-α + Th1 cells in peripheral blood and liver tissues decreased after hPMSCs treatment ( P<0.001), but the expression of PD-1 on this T cell subset was promoted in peripheral blood and liver tissues ( P<0.01, P<0.001). However, the intervention effects of shCD73 on TNF-α + Th1 cells in peripheral blood and liver tissues were significantly weakened ( P<0.05, P<0.01). Liver histopathological analysis showed that the proportion of TNF-α + Th1 cells in liver was positively correlated with Suzuki′s score, collagen area and the MFI of α-SMA ( P<0.001). Similarly, histopathological analysis of skin and lung tissues also showed that the proportion of TNF-α + Th1 cells in peripheral blood was positively correlated with skin Marina score and lung Shukai Qiao score ( P<0.001). In vitro experiment also showed that hPMSCs down-regulated the proportion of TNF-α + Th1 cells ( P<0.01) and up-regulated the expression of PD-1 on them ( P<0.05). Further analysis showed that hPMSCs could enhance the MFI of Nrf2 ( P<0.05) and weaken the MFI of p-NF-κB ( P<0.01) in TNF-α + Th1 cells. Conclusions:hPMSCs could up-regulate the expression of PD-1 through CD73/Nrf2 pathway to inhibit the formation of TNF-α + Th1 cells, thereby alleviating liver injury in GVHD mice.

Correlation of neutrophil extracellular traps with T cells differentiation in Hashimoto′s thyroiditis mice and vitamin D′s relief effect / 中华内分泌代谢杂志

Objective:To investigate correlation between neutrophil extracellular traps(NETs) formation and T cell subsets in mice with experimental autoimmune thyroiditis(EAT) and the impact of active vitamin D intervention.Methods:Six-week-old female BALB/c mice were randomly divided into Control group, EAT group and 1, 25 dihydroxy vitamin D 3[1, 25(OH) 2D 3] treatment group(VitD group; n=6/group). HE staining was used to observe thyroid pathology. Plasma thyroglobulin antibody(TGAb), thyroid peroxidase antibody(TPOAb), and 1, 25(OH) 2D 3 were measured by ELISA. Peripheral NETs formation, Th1, Th2, and Th17 cell ratio from spleen were measured by flow cytometry. Correlation between NETs formation rate and Th1, Th2, and Th17 cell ratio was analyzed. Results:Compared with Control group, mice in EAT group had significantly increased thyroid inflammation scores, thyroiditis morbidity, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + )and Th17 cell proportions( P were <0.001, 0.002, 0.007, <0.001, <0.001, 0.003, 0.001, and 0.002, respectively), and significant decreased 1, 25(OH) 2D 3, Th1 cell proportions, Th1/Th2(CD4 + IL-4 + ), Th1/Th2(CD4 + IL-13 + ), and Th1/Th17 ratios( P were 0.010, 0.018, 0.010, 0.005, and 0.007, respectively). Compared with the EAT group, the VitD group had lower thyroid inflammation scores, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportions( P were 0.044, 0.007, <0.001, 0.001, 0.014, 0.008, and 0.001, respectively), and significant higher Th1 cell ratio, Th1/Th2(CD4 + IL-13 + ) and Th1/Th17 ratio( P were 0.011, 0.009, and 0.003, respectively). The Th1/Th2(CD4 + IL-4 + ) was not significantly increased in VitD group compared with EAT group( P=0.174). NETs formation rate was positively correlated with Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportion( r were 0.65, 0.59, and 0.61; and P were 0.004, 0.010, and 0.007, respectively), but not with Th1 cell proportion( r=-0.47, P=0.051). Conclusion:EAT mice were more prone to NETs formation. Active vitamin D may relieve immune imbalance with increased Th2 and Th17 cell ratio and decreased Th1 cell ratio by reducing the formation of NETs in EAT mice. Vitamin D played the protective role in thyroid by reducing thyroid pathological damage and thyroid autoantibody levels, and relived overall lymphocyte imbalance.

Effect of T helper 1 to T helper 2 ratio on the prognosis of patients with multiple myeloma / 白血病·淋巴瘤

Objective:To investigate the effect of T helper 1 (Th1) to T helper 2 (Th2) ratio (Th1/Th2) on the prognosis of patients with multiple myeloma (MM).Methods:The clinical data of 168 MM patients who were newly diagnosed in the Affiliated Drum Tower Hospital of Nanjing University Medical School from January 2016 to January 2021 were retrospectively analyzed. Disease staging was defined according to the Chinese guidelines for diagnosis and treatment of MM (2020 edition). Risk stratification was based on the Mayo stratification of myeloma and risk-adapted therapy (mSMART) 3.0. The levels of Th1 and Th2 in peripheral blood of patients were detected by flow cytometry. Th1/Th2 was compared among patients with different disease staging and risk stratification. Using mSMART 3.0 risk stratification as the gold standard, the receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value of Th1/Th2 for determining high-risk MM. According to the optimal cut-off value, patients were divided into high Th1/Th2 group (≥ optimal cut-off value) and low Th1/Th2 group (< optimal cut-off value). The progression-free survival (PFS) of the two groups was analyzed by Kaplan-Meier method. Multivariate Cox proportional hazards model was used to analyze the influencing factors of PFS.Results:There were 40, 62 and 66 patients with international staging system (ISS) stages Ⅰ, Ⅱ and Ⅲ, respectively, with Th1/Th2 [ M ( IQR)] of 19.20 (18.98), 15.93 (14.40) and 14.47 (12.01), respectively ( H = 6.68, P = 0.036). There were 31,102 and 35 patients with revised international staging system (R-ISS) stages Ⅰ, Ⅱ and Ⅲ, respectively, with Th1/Th2 of 19.67 (21.92), 14.87 (11.36) and 13.50 (12.80), respectively ( H = 7.26, P = 0.027). There were 99 and 69 patients with mSMART 3.0 high-risk and standard-risk, respectively, and the Th1/Th2 of high-risk patients was lower than that of the standard-risk patients [14.70 (11.93) vs. 17.72 (16.80), U = 2 612.00, P = 0.009]. ROC curve analysis showed that the area under the curve for Th1/Th2 to determine high-risk MM was 0.618 (95% CI 0.531-0.705, P = 0.010), and the optimal cut-off value was 16.55 and there were 81 and 87 cases in the high Th1/Th2 group and low Th1/Th2 group. With a median follow-up of 28 months (1-70 months), the median PFS time for all patients was 36 months (95% CI 29-43 months); PFS in high Th1/Th2 group was better than that in low Th1/Th2 group [median PFS time: 39 months (95% CI 26-51 months) vs. 28 months (95% CI 21-34 months), P = 0.040]. Multivariate Cox regression analysis showed that renal impairment (with vs. without: HR = 2.340, 95% CI 1.350-4.053, P = 0.002) and low Th1/Th2 (high vs. low: HR = 0.551, 95% CI 0.344-0.882, P = 0.013) were independent risk factors for PFS in MM patients. Conclusions:The imbalance between Th1 and Th2 is associated with the prognosis of MM patients, and patients with low Th1/Th2 are at high risk of progression. Th1/Th2 can be used as a prognostic indicator for MM.

The relationship between C3, C4, Th1/Th2 levels and the MG-ADL score in patients with myasthenia gravis and its efficacy in predicting the transition of ocular muscle type to systemic type / 中国医师杂志

Objective:To investigate the relationship between C3, C4, Th1/Th2 levels and the Myasthenia Gravis Daily Living Scale (MG-ADL) score in patients with myasthenia gravis (MG) and its efficacy in predicting the transition of ocular muscle type to systemic type.Methods:A retrospective study of 94 patients with ophthalmic MG admitted to Haikou People's Hospital from April 2017 to April 2020 was conducted. According to whether they had converted to systemic MG within 6 months, they were divided into transformation group ( n=35) and non-transformation group ( n=59). The levels of C3, C4 and Th1/Th2, as well as the score of MG-ADL and Quantitative Myasthenia Gravis (QMG) were compared between the two groups before and 1 and 3 months after treatment. The correlation between C3, C4 and Th1/Th2 levels and MG-ADL and OMG scores, as well as the related influencing factors of the transformation from ocular muscle type to systemic type was analyzed. The efficiency of each index in predicting the transformation from ocular muscle type to systemic type was analyzed. Results:At 1 and 3 months after treatment, the C3 and C4 in both groups were significantly higher than before treatment, and Th1/Th2 was significantly lower than before treatment; the C3 and C4 in the non-transformation group were higher than that in the transformation group, while Th1/Th2 was lower than that in the transformation group (all P<0.05). The MG-ADL and QMG scores in 2 groups at 1 and 3 months after treatment were significantly lower than those before treatment, and those in the non-transformation group were lower than those in the transformation group (all P<0.05). C3 and C4 levels were negatively correlated with MG-ADL and QMG scores (all P<0.05), while Th1/Th2 levels were positively correlated with MG-ADL and QMG scores (all P<0.05). At 1 and 3 months after treatment, C3, C4 and Th1/Th2 were the influencing factors for the transformation from ocular muscle type to systemic type (all P<0.05). The area under the curve (AUC) of C3, C4 and Th1/Th2 combined to predict the transformation from ocular muscle type to systemic type at 3 months after treatment was 0.939, and the best predictive sensitivity and specificity were 91.43% and 88.14%, respectively. Conclusions:There is a good linear relationship between C3, C4, Th1/Th2 levels and MG-ADL scores in MG patients, and it has a high efficiency in predicting the transition of ocular muscle type to systemic type.

Significance of Th1/Th2 cytokines in prognostic stratification of acute myeloid leukemia / 白血病·淋巴瘤

Objective:To investigate the significance of Th1/Th2 cytokines in prognostic stratification of acute myeloid leukemia (AML).Methods:A total of 83 patients with newly diagnosed AML from June 2017 to April 2019 in the First People's Hospital of Yunnan Province were collected. According to the Chinese guidelines for diagnosis and treatment of adult acute myeloid leukemia (non-acute promyelocytic leukemia) (2017 edition), AML patients were divided into poor prognosis group (45 cases), moderate prognosis group (19 cases), and good prognosis group (19 cases); moderate prognosis plus poor prognosis was treated as the not good prognosis. Mann-Whitney U test and Kruskal-Wallis H test were used to compare the expression differences of Th1/Th2 cytokines in peripheral blood of different prognosis groups; cytokines with statistical differences among different prognosis groups were selected, and the cut-off value of AML patients with different prognostic stratification distinguished by cytokines was determined by using receiver operating characteristic (ROC) curve. Finally, patients were divided into ≥ cut-off value group and <cut-off value group according to the cut-off value, and then the association of both groups with the prognostic stratification in guideline was also analyzed. Results:The median expression level of tumor necrosis factor (TNF)-β of patients in moderate prognosis group [3.80 pg/ml (2.75 pg/ml, 15.32 pg/ml)] was higher than that of patients in poor prognosis group [2.78 pg/ml (1.28 pg/ml, 3.36 pg/ml)] and good prognosis group [1.61 pg/ml (0.83 pg/ml, 3.04 pg/ml)] ( U=216, P=0.02; U = 312, P < 0.05); the median expression level of TNF-β in good prognosis group was lower than that in poor prognosis group ( U = 562, P = 0.048). There were no statistically significant differences in the expression levels of Th1/Th2 cytokines of AML patients with different prognostic stratification (all P>0.05).The cut-off value of TNF-β was 3.23 pg/ml in good prognosis group and moderate prognosis group, the area under the ROC curve was 0.866 (95% CI 0.753-0.978, P < 0.05); among 26 patients with TNF-β≥ 3.23 pg/ml, 25 (96.2%) patients had not good prognosis. The cut-off value was 3.62 pg/ml for distinguishing between moderate prognosis group and poor prognosis group, the area under the ROC curve was 0.747 (95% CI 0.610-0.884, P = 0.02); 18 (100%) patients with TNF-β≥ 3.62 pg/ml had not good prognosis. The cut-off value was 2.19 pg/ml for distinguishing between good prognosis group and not good prognosis group, the area under the ROC curve was 0.719 (95% CI 0.595-0.842, P = 0.04); among 53 patients with TNF-β≥2.19 pg/ml, 46 (86.8%) patients had not good prognosis. Conclusions:The high expression of TNF-β may indicate that the prognosis of AML patients is not good. When the level of TNF-β was equal or greater than 3.62 pg/ml, it may contribute to the prognostic stratification of AML patients.

Mechanism of / 南方医科大学学报

OBJECTIVE@#To explore the mechanism of @*METHODS@#Healthy male DBA/1 mice were used for CIA modeling. Twenty-five CIA mice with successful modeling and similar arthritis index (AI) scores were randomized equally into model group (CIA), methotrexate (MTX) group, and low-, medium-, and high-dose XWGD groups (0.975, 1.95, and 3.9 g/mL, respectively), with another 5 normal mice as the normal control group. The mice in normal control and CIA groups were given saline once a day, those in MTX group were given 0.1 mg/mL MTX once a week, and those in XWGD groups were treated daily via garage of XWGD containing crude drugs of different doses for 28 consecutive days. The AI score and HE staining were used to evaluate the changes in the joints of the CIA mice. The effect of XWGD on Th1, Th17, MDSC, G-MDSC and M-MDSC cells were evaluated with flow cytometry.@*RESULTS@#Treatment with MTX and different doses of XWGD significantly decreased the AI score of the mice and relieved joint inflammation as compared with the model group (@*CONCLUSIONS@#XWGD can improve joint inflammation in CIA mice by increasing the percentages of G-MDSC cells and decreasing the percentages of M-MDSC, Th1 and Th17 cells, and a high dose of XWGD can produce an equivalent therapeutic effect to methotrexate but with better safety.

Effect of epigallocatechin gallate on Th1/Th2 immune balance in patients with plaque-type psoriasis / 中华皮肤科杂志

Objective@#To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.@*Methods@#A total of 33 patients with plaque-type psoriasis vulgaris were enrolled, and peripheral blood mononuclear cells (PBMC) were isolated and cultured. The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay. PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours. Flow cytometry was performed to determine proportions of Th1 and Th2 cells, enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin[IL]-2, interferon[IFN]-γ) and Th2 cytokines (IL-4, IL-10) in the cell culture supernatant, and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor) . Statistical analysis was carried out by using t test.@*Results@#According to the MTT assay results, EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments. Compared with the control group, the EGCG group showed significantly decreased number of Th1 cells (t = 3.43, P = 0.026) , increased number of Th2 cells (t = 6.68, P = 0.026) , and decreased Th1/Th2 ratio (P < 0.05) . The levels of IL-2 and IFN-γ in the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L, 1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ± 196.45 ng/L, 3 287.63 ± 235.54 ng/L respectively) , while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L, 285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L, 165.46 ± 32.25 ng/L respectively) . Compared with the control group, the EGCG group showed significantly decreased T-bet mRNA expression (t = 11.99, P < 0.001) , but increased GATA3 mRNA expression (t = 18.62, P < 0.001) .@*Conclusion@#EGCG can reduce the number of Th1 cells, inhibit the production of Th1 cytokines and transcription factors, and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors, followed by the modulation of Th1/Th2 immune imbalance.

The role of Th1 and Th2 cells in the severity of acute pancreatitis / 中华急诊医学杂志

Objective To investigate the relationship between the levels of Thl and Th2 cells and their cytokines IFN-γ,TNF-αt,IL-2,IL-4,IL-5 and IL-13 in peripheral blood of patients with acute pancreatitis and the clinical severity of acute pancreatitis,and to evaluate the diagnostic value and significance for the severity of acute pancreatitis.Methods This study enrolled 72 patients with acute pancreatitis and 30 healthy controls admitted to the Emergency Surgery Department of the First Affiliated Hospital of Zengzhou University from January 2015 to June 2017.The patients were divided into the mild MAP group,moderate MSAP group,and severe SAP group according to the 2012 Atlanta classification criteria.Whole blood and serum samples were taken on the day of admission for all confirmed cases,the number of Th1 and Th2 cells was detected by flow cytometry,the expression levels of IFN-γ,TNF-α,IL-2,IL-4,IL-5 and IL-13 were determined by ELISA.The total length of hospital stay and length of hospital stay in ICU of each group were recorded.The number of Th 1 and Th2 cells and the expression of related cytokines in patients with mild,moderate and severe acute pancreatitis were compared by one-way analysis of variance.Pearson linear correlation was used to analyze the correlation between Th1and Th2 cytokines and the length of hospital stay and the length of hospital stay in ICU.A P＜0.05 was considered statistically significant.Results Compared with the healthy controls,the number of Th1 and Th2 cells,the levels of Th1 cytokine IFN-γ,TNF-α IL-2 and Th2 cytokine IL-4 were significantly increased in acute pancreatitis.Th1 cells in the SAP group were significantly higher than those in the MAP and MSAP groups (F=11.137,P＜0.01),while Th2 cells in the SAP group were significantly lower than those in the MAP and MSAP groups (F=9.493,P＜0.01),and there was no significant difference between the MAP group and MSAP group.The expression level of IFN-γ was higher in the SAP and MSAP groups than that in the MAP group (F=8.605,P=0.001).The expression level of TNF-α was higher in the SAP group than those in the MSAP and MAP groups (F=1 1.847,P＜0.01),and the IL-4 expression level was significantly lower in the SAP group than those in the MAP and MSAP groups (F=8.042,P=0.001).The expression level of IFN-γ was positively correlated with the length of hospital stay and the length of hospital stay in ICU (r=0.569,P=0.014;r=0.538,P=0.021).The expression level of TNF-α was positively correlated with the length of hospital stay (r=0.475,P=0.046).The expression level of IL-4 was negatively correlated with the length of hospital stay and the length of hospital stay in ICU (r==0.577,P=0.012;r=-0.657,P=0.003).Conclusions Th1 and Th2 cells and their associated inflammatory factors IFN-γ,TNF-α,IL-2 and IL-4 are elevated in patients with acute pancreatitis.The increase in the number of Th1 proinflammatory cytokines IFN-γ,TNF-α and Th1 cells tends to aggravate the development of acute pancreatitis,and the increase in the number of Th2 cells and the anti-inflammatory cytokine IL-4 are associated with a better prognosis.

The changes of perioperative immunity index in patients with breast cancer and its clinical significance / 中华肿瘤杂志

Objective@#To investigate the changes of perioperative immune index in patients with breast cancer and its clinical significance.@*Methods@#Th1 cells, Th2 cells, Th1/Th2 ratio and regulatory T cells (Treg) were detected in peripheral blood of 103 patients with primary breast cancer and 116 patients with breast fibroma before surgery and on the 1st, 3rd and 5th day following operation. The relationship of changes in T lymphocyte subsets and clinicopathological characteristics, as well as tumor-free survival of breast cancer patients, was analyzed.@*Results@#The levels of Th1 cells in breast cancer group on the 1st, 3rd and 5th day following operation were (12.20±0.45)%, (13.89±0.47)%, (14.04±0.49)%, which were significantly lower than those before operation [(15.82 + 0.51)%, all P<0.05 ]. Treg cells, however, with the number of (3.82±0.13)%, (3.25±0.11)%, (2.95 ±0.11)%, were remarkably higher than those before operation [(2.53 ±0.11)%, all P<0.05]. With respect to breast fibroma patients, there was no significant difference compared with those before operation of Th1 cells, Th2 cells and Treg cells (all P>0.05). The changes of Th1 cells were associated with the degree of differentiation, T stage, N stage, TNM stage, HER-2 status and Ki-67 (all P<0.05). Treg cells were related to T stage, N stage and HER-2 status (all P<0.05). Tumor-free survival in the Th1-cell-increasing group was significantly better than that in the Th1-cell-decreasing group (P=0.045), while cell-decreasing group of Treg showed the improved outcomes (P=0.012).@*Conclusions@#The levels of Th1 cells and Treg cells are important indicators of cellular immune function in patients with breast cancer. Moreover, the perioperative changes of Th1 cells and Treg cells are associated with the size of tumors, pathological parameters, clinical stages and tumor-free survival outcomes.

Research advances in Th1/Th2 balance and fibrotic diseases / 临床肝胆病杂志

Body immunity is closely associated with the pathogenesis of fibrotic diseases. In CD4+ T lymphocyte subsets, T helper 1 (Th1)/T helper 2 (Th2) balance plays an important regulatory role in the development and progression of many diseases. This article describes the association of Th1/Th2 balance with liver fibrosis, pulmonary fibrosis, and renal fibrosis. It is believed that in addition to the measurement of single cytokines, further studies are needed to clarify the specific association between T helper cells and other cells and related mechanisms.

Effect of epigallocatechin gallate on Th1/rh2 immune balance in patients with plaque-type psoriasis / 中华皮肤科杂志

Objective To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.Methods A total of 33 patients with plaque-type psoriasis vulgaris were enrolled,and peripheral blood mononuclear cells (PBMC) were isolated and cultured.The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay.PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours.Flow cytometry was performed to determine proportions of Th 1 and Th2 cells,enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin [IL]-2,interferon [IFN]-γ) and Th2 cytokines (IL-4,IL-10) in the cell culture supernatant,and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor).Statistical analysis was carried out by using t test.Results According to the MTT assay results,EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments.Compared with the control group,the EGCG group showed significantly decreased number of Th1 cells (t =3.43,P =0.026),increased number of Th2 cells (t =6.68,P =0.026),and decreased Th1/Th2 ratio (P ＜ 0.05).The levels of IL-2 and IFN-γin the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L,1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ±196.45 ng/L,3 287.63 ± 235.54 ng/L respectively),while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L,285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L,165.46 ± 32.25 ng/L respectively).Compared with the control group,the EGCG group showed significantly decreased T-bet mRNA expression (t =11.99,P ＜ 0.001),but increased GATA3 mRNA expression (t =18.62,P ＜ 0.001).Conclusion EGCG can reduce the number of Th1 cells,inhibit the production of Th 1 cytokines and transcription factors,and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors,followed by the modulation of Th 1/Th2 immune imbalance.

Artesunate promotes Th1 differentiation from CD4+ T cells to enhance cell apoptosis in ovarian cancer via miR-142

The aim of this study was to evaluate the influence of artesunate on Th1 differentiation and its anti-tumor effect on ovarian cancer. A Murine ovarian cancer model was established by ID8 cells transplantation. The expression of miR-142 and Sirt1 proteins in peripheral CD4+ T cells were quantified with qRT-PCR and western blot, respectively. Peripheral CD4+ T cells were induced for Th1 differentiation. The percentages of apoptosis of Th1/CD4+ T cells and ovarian cancer cells were analyzed by flow cytometry. The IFN-γ level was examined through enzyme-linked immunosorbent assay. Artesunate promoted miR-142 expression in peripheral CD4+ T cells and Th1 differentiation from CD4+ T cells. Artesunate promoted cell apoptosis of ovarian cancer cells by inducing Th1 differentiation. By up-regulating miR-142, artesunate suppressed Sirt1 level and promoted Th1 differentiation. Artesunate enhanced the pro-apoptotic effects of Th1 cells on ovarian cancer via the miR-142/Sirt1 pathway. Artesunate promoted Th1 differentiation from CD4+ T cells by down-regulating Sirt1 through miR-142, thereby enhancing cell apoptosis in ovarian cancer.

Study on the expression and significance of Th1/Th2 cells in the blood of patients with primary gouty arthritis / 中华风湿病学杂志

Objective This study is aimed to investigate the possible role of Th1/Th2 cell in the pathogenesis of primary gout arthritis. Methods The peripheral blood of 21 acute gout patients (AG), 20 intermittent gout patients (IG) and 20 healthy controls (HC) were collected. The clinical data and laboratory indicators of them were enrolled. The percentages of Th1 and Th2 cells were detected by flow cytometry (FCM). The expression of GATA-3, T-bet, IL-4 and interferon (IFN)-γ mRNA in Peripheral blood mononuclear cells (PBMCs) were measured using Real time quantitative polymerase chain reaction (PCR). The protein expression levels of IL-4 and IFN-γ in serum were detected by enzyme-linked immuno sorbent assay (ELISA). The measurement data were compared by one factor analysis of variance test. The correlation between variables was used by Spearman correlation analysis. Results The percentage of Th1 cells in peripheral blood of the AG group was [(23.2 ±8.3)％], and the IG group was [(20.5 ±9.3)％], which were significantly higher (F=6.520, P<0.05) than the percentage of HC group [(14.8±3.8)％]. There was no significant difference between AG group and IG group. The percentage of Th2 cells in peripheral blood of AG group were [(1.9 ±0.7)％], which was significantly lower (F=8.267, P<0.05) than and the percentage of HC group [(3.4±1.8)％] and IG group [(3.3± 1.2)％]. There was no significant difference between the IG group and the HC group. And the proportion of TH1/Th2 cells in the AG group was higher than that of the IG group and the HC group (F=10.406, P<0.01). The expression of T-bet mRNA and IFN-γ mRNA in the AG group and IG group were higher than that in the HC group (F=4.942, P=0.010)、(F=4.458, P=0.016). However, there was no significant difference between IG group and AG group. The expression of GATA-3 mRNA and IL-4 mRNA was significantly lower (F=3.564, P=0.035) (F=5.385, P=0.007) in the AG group and IG group when compared to the HC group. And there was no significant difference between the AG and the IG group. The IFN-γ level increased in the AG and IG group compared to the HC group (F=7.659, P=0.001). The IL-4 levels in the AG group was lower (F=7.099, P=0.002) than those of the IG and HC group. Conclusion Th1 cells in the peripheral blood of patients with GA are increased and accompanied with the decrease of Th2 cells. The results of this study suggest that the imbalance of Th1/Th2 cells in the inflammatory and immune response plays a critical role in the pathogenesis of GA.

Serosal Cavities Contain Two Populations of Innate-like integrin α4highCD4+ T Cells, Integrin α4β1+α6β1+α4β7− and α4β1+α6β1−α4β7+ Cells

We previously reported peritoneal innate-like integrin α4 (CD49d)highCD4+ T cells that provided help for B-1a cells. Here we analyzed the expression of various integrin chains on the peritoneal and pleural integrin α4highCD4+ T cells and investigated the functional heterogeneity of the subpopulations based on the integrin expression. Pleural cavity contained a lower ratio of integrin α4highCD4+ T cells to integrin α4lowCD4+ T cells than peritoneal cavity, but the pleural integrin α4highCD4+ T cells have the same characteristics of the peritoneal integrin α4highCD4+ T cells. Most of integrin α4highCD4+ T cells were integrin β1highβ7−, but a minor population of integrin α4highCD4+ T cells was integrin β1+β7+. Interestingly, the integrin α4highβ1highβ7− CD4+ T cells expressed high levels of integrin α4β1 and α6β1, whereas integrin α4highβ1+β7+ CD4+ T cells expressed high levels of integrin α4β1 and α4β7, suggesting an alternative expression of integrin α6β1 or α4β7 in combination with α4β1 in respective major and minor populations of integrin α4highCD4+ T cells. The minor population, integrin α4highβ1+β7+ CD4+ T cells, were different from the integrin α4highβ1highβ7− CD4+ T cells in that they secreted a smaller amount of Th1 cytokines upon stimulation and expressed lower levels of Th1-related chemokine receptors CCR5 and CXCR3 than the integrin α4highβ1 highβ7− CD4+ T cells. In summary, the innate-like integrin α4highCD4+ T cells could be divided into 2 populations, integrin α4β1+α6β1+α4β7− and α4β1+α6β1−α4β7+ cells. The functional significance of serosal integrin α4β7+ CD4+ T cells needed to be investigated especially in view of mucosal immunity.

Changes of serum Th1/Th2 cytokines in peripheral blood pre-and post-liver transplantation / 天津医药

Objective To explore the trend of changes of Th1 and Th2 cytokines within 6 months after adult liver transplantation (LT). Methods Twenty-three patients from Tianjin First Center Hospital were chosen as the patient group with an average age of (52.7 ± 7.6), range from 37 to 63 years old, including 21 males and 2 females. Twenty healthy staffs from Tianjin First Center Hospital formed the control group (C) with 15 males and 2 females whose average age was (31.0 ± 6.1) ranged from 22 to 24 years old. The patient group was treated with tacrolimus after LT as main immunosuppressive drug. The peripheral blood at time points before (T0) and 1 month (T1), 3 months (T3), 6 months (T6) after LT at 9:00 AM were collected. The blood sample was also collected form control group but only one time. Levels of IL-2, IFN-γ, IL-10 and TGF-βwere detected by ELISA. Results (1) The concentration of IL-2 showed a continuous up-going trend, which was not such obvious between T1 and T0, and until T3 reached a higher concentration than T0. The concentration at T6 was higher than T0 and T1. There were no significant differences in concentrations of T0 to T3 between patient group and control group, while T6 reached a higher concentration in patient group than that of the control group. (2) The concentration of IFN-γexperienced a shortly down-going trend from T0 to T3, and started rising, reached the peak at 3 months after the operation, then started its down-going trend. There were no significant differences in the concentrations of IFN-γfrom T1 to T6, and T3 reached a higher concentration than T1 while T6 was lower than T3. Only at T3, the concentration of IFN-γwas higher in patient group than that of control group. (3) There were no significant differences in the concentrations of IL-10 at various time points in patient group, and there were no significant differences in the concentrations of IL-10 at different time points between two groups (P>0.05). (4) The concentration of TGF-βshowed a gradual decline after the operation, and reached its bottom at T6, and which was lower than T0 to T3. Compared with the control group, the down-going trend was not such obviously at T0 and T1, and the concentration was down at T3 and T6(P<0.05). Conclusion Our results suggest that there is a tendency of an increasing Th1 cytokine expression at early stage in post-transplantation, while the TGF-βof Th2 cytokine is a decreasing trend. This tendency may associate with the autoimmunity response caused by LT and the immunosuppressive drugs.

Interpretation of laboratory test results in early and late onset severe preeclampsia / 中华检验医学杂志

Objective To analyze the difference of laboratory test results between early-onset and late-onset severe preeclampsia and to investigate their clinical application values.Methods Totally 108 blood samples were collected from patients with severe preeclampsia who were diagnosed according to the Diagnostic Standard of Obstetrics and Gynecology(7th Edition) published by People′s Medical Publishing House,in Shandong Provincial Hospital affiliated to Shandong University from March to November 2016,which consisted of 64 early-onset severe preeclampsia before 34 weeks gestation(early onset group) and 44 late-onset severe preeclampsia after 34 weeks gestation(late onset group).In addition,42 women with normal pregnancies as the control group were selected.General clinical data were collected,and the blood sample was analyzed through detecting Hb,PLT,fibrinogen (FIB),D-dimer,AST,ALT,urea,creatinine (Cr),uric acid,CRP,urine protein.The tested results were analyzed and compared.Flow cytometry was used to analyze the proportion of T helper 1 cells(Th1) and T helper 2 cells(Th2),and the ratio of Th1/Th2 was also calculated.All data and F test were performed by use of statistical software SPSS19.0.Results The pre-pregnancy body mass index(29.55±4.49,30.66±5.13,26.62±3.17,F=9.829,P<0.05),diastolic blood pressure[(105.17±14.46)mmHg(1 mmHg=0.133 kPa),(99.80±12.56)mmHg,(74.36±8.42)mmHg,F=82.088,P<0.05],Hb[(123.22±14.38)g/L,(117.03±16.48)g/L,(112.62±11.24)g/L,F=7.133,P<0.05],urea[(6.56±2.36)mmol/L,(4.51±1.35)mmol/L,(3.04±0.87)mmol/L,F=51.733,P<0.05],Cr[(68.47±18.05)μmol/L,(61.37±14.37)μmol/L,(48.54±8.73)μmol/L,F=23.737,P<0.05],CRP[(7.68±8.76)mg/L,(5.88±6.03)mg/L,(3.56±2.41)mg/L,F=4.735,P<0.05],urine protein[(3.66±0.76)g/L,(2.20±1.05)g/L,(0.19±0.40)g/L,F=249.714,P<0.05]had a statistically significant difference among the early-onset,late-onset and control groups.The flow cytometry results demonstrated that the proportion of Th1 in early-onset group(19.83±3.04)was higher than that in both late-onset (14.49±2.79)and control groups(11.78±1.17),on the contrary,the result of Th2 was much lower(early-onset:1.02±0.12,late-onset: 1.11±0.12,control: 1.56±0.11),there was statistical significance among these three groups(Th1: F=135.110,P<0.05;Th2: F=293.687,P<0.05).Conclusions It′s necessary to real-time monitor the laboratory indicators,such as liver and kidney function,especially the immunologic function indicators for evaluating the disease of early-onset and late-onset severe preeclampsia and personal treatment,and for ensuring the health of mother and fetus and improving the prognostic of mother and fetus.

Study on the correlation of the effect of entecavir on Th1/Th2 cytokines level in the treatment of chronic hepatitis / 中华肝脏病杂志

Objective@#To explore the expression level of peripheral blood Th1/Th2 type cytokines of chronic hepatitis b (CHB) patients in the entecavir (ETV) antiviral treatment, analyze the relationship between various cytokines, and the correlation among of cytokines and HBV DNA loads.@*Methods@#Luminex Liquid Chip Technology was applied to detect the peripheral blood Th1/Th2 type cytokines expression level of CHB patients; At the same time, liver function was detected by Fully Automatic Biochemical Analyzer; HBV DNA loads were detected by PCR Method; Hepatitis b virology markers were detected by Chemiluminescence Method. F-test and Pearson correlation analysis were used for statistical analysis.@*Results@#Before ETV antiviral treatment, peripheral blood Th1 cytokines IFN gamma expression level in patients with CHB increased significantly (P = 0.010) compared with the healthy control group while TNF alpha expression level having no statistically significant difference (P = 0.095); Th2 type cytokines IL-4, IL-6, IL-10 levels decreased obviously (P = 0.039, P = 0.014, P = 0.026) compared with those in the control group. After 48 weeks of treatment, Th1 cytokines IFN gamma and TNF alpha expression levels were reduced significantly (19.2±5.03 pg/ml vs 24.69±6.51 pg/ml, and 6.09±4.99 pg/ml vs 9.50±7.34 pg/ml, P < 0.001, and P = 0.016) compared with that before treatment; Th2 type cytokines IL-4, IL-6 expression level increased significantly (33.86±22.47 pg/ml vs 21.32±12.84 pg/ml, and 11.65±6.91 pg/ml vs 8.51±4.94 pg/ml, P = 0.004, and P = 0.029) compared with that before treatment while IL-10 expression level having no statistical significance (P = 0.081). There was disorder and irregularity in the correlation between the levels of Th1/Th2 type cytokines. And there was no correlation between the various cytokines and HBV DNA loads in patients with CHB.@*Conclusion@#ETV can not only inhibit HBV DNA replication, reducing HBV DNA loads, but also contribute to regulate Th1/Th2 type cytokines expression level in patients with CHB, but there was no correlation between the levels of various cytokines, various cytokines and HBV DNA loads.

Effect of immature dendritic cells (imDC) induced by sinomenine on the peripheral blood Th1/Th2 cytokines of allogeneic rat renal transplantation / 中国医师杂志

Objective To investigate the effect of immature dendritic cell (imDC) derived from donor-derived bone marrow induced by alkaloid sinomenine (SN) on the Th1/Th2 cytokines in venous blood of receptors,and to probe into the mechanism which imDC induced by SN can lead transplantation immune tolerance.Methods Inbred strain Wistar and Sprague Dawley (SD) rats were selected as kidney transplant donor and recipient,respectively.Vessel sutures of the microsurgery technique were used to build the bilaterally renal transplantation model of rats.By the injection of imDC to the recipient rats preoperatively,enzyme-linked immunosorbent assay (ELISA) was used to determine the level of the interleukin (IL)-2,IL-4,IL-10 and interferon-γ (INF-γ).Results (1) Successful rate of transplantation was 89.5％.Arterial anastomosis time was (12.5 ±5.7)min,and venous anastomosis time was (17.3 ± 3.4)min.(2) The content of the IL-2,INF-γ,IL-4 and IL-10 in SN-imDC 106 group was (17.25 ± 3.41) pg/ml,(239.80 ± 9.06) pg/ml,(337.60 ± 25.07) pg/ml,and (1 432.00 ± 106.39) pg/ml,respectively.Among the same concentration,the level of the IL-4 and IL-10 that stood for the Th2 cytokines was significantly higher in SN-imDC group than imDC group and control group (P ＜ 0.05),and was significantly higher in SN-imDC 106 group than SN-imDC 105 group (P ＜ 0.05).Among the same concentration,the lever of IL-2 and INF-γ that stood for the Th1 cytokines was significantly lower in SN-imDC group than imDC group and control group (P ＜ 0.05),and was significantly lower in SN-imDC 106 group than SN-imDC 105 group (P ＜ 0.05).Conclusions (1) The use of microsurgery for anastomosis could make the model of singel kidney transplantation in rats.(2) Specific imDC induced by SN could induce the migration to Th2 immune response,which proved imDC induced by SN could mediate immune tolerance to the recipient.

Imbalance among Th1, Th2 and Th17 Cells and Crohn's Disease / 胃肠病学

Dysregulated immune response is crucial for the pathogenesis of inflammatory bowel disease.Recent studies suggested that imbalance among Th1, Th2 and Th17 cells was closely related to the aberrant intestinal immune response.Aims: To investigate the association of imbalance among Th1, Th2 and Th17 cells and Crohn''s disease (CD).Methods: Thirty-six CD patients admitted from Jan.2013 to Dec.2014 at the Second Affiliated Hospital of Wenzhou Medical University were enrolled;40 patients undergoing intestinal polypectomy were collected as controls.Inflamed and normal mucosal tissues were obtained from CD patients and the controls, respectively;expressions of Th1, Th2 and Th17 cells related transcriptional factors (T-bet, GATA-3 and RORγt) and cytokines (IFN-γ, IL-4 and IL-17A) were determined by real-time PCR and immunohistochemistry.Results: In comparison with the controls, mRNA expression of T-bet and RORγt, mRNA and protein expressions of IFN-γ and IL-17A, as well as the ratios for T-bet/GATA-3 and RORγt/GATA-3, which represented balance of Th1/Th2 and Th17/Th2, respectively, were all significantly increased in inflamed mucosal tissue in CD patients (P<0.05).Expression of each of the three transcriptional factors was positively correlated with its corresponding cytokine (P<0.05).IFN-γ+ and IL-17A+ cells mainly located in the intestinal epithelial layer and lamina propria with cytoplasmic immunoreactivities in CD patients.In the stratified analysis, all the above-mentioned parameters were significantly higher in active CD than in inactive CD (P<0.05);furthermore, the ratio for RORγt/T-bet, which represented balance of Th17/Th1, was also increased significantly in active CD (P<0.05).Conclusions: Imbalance among Th1, Th2 and Th17 cells in intestinal mucosal tissue was closely related with CD.Polarization of Th1 and Th17 cells are involved in this process, and Th17 polarization is predominant in active disease.

Citocinas de resposta Th1 e Th2 e diabetes mellitus tipo 1 / Th1 and Th2 response cytokins in type 1 diabetes mellitus

O diabetes mellitus tipo 1 (DM1) é uma doença autoimune crônica, onde as células T autorreativas destroem as células beta pancreáticas levando à dependência de insulina exógena. Para o desenvolvimento do DM1 são necessárias numerosas interações entre as células do sistema imunológico, principalmente mediadas por citocinas de resposta T helper 1 (Th1) e T helper 2 (Th2). O presente estudo teve como objetivo analisar a relação das citocinas de resposta Th1 e Th2 no desenvolvimento do DM1 por meio de uma revisão de literatura, avaliando artigos científicos eletrônicos publicados entre os anos de 2001 e 2016, além de livros de imunologia aplicados à clínica. Diversos estudos na literatura demonstram que o perfil de secreção de citocinas durante o desenvolvimento do DM1 é de padrão Th1 onde temos como principais constituintes a IL-2 e o IFN-y. Já as citocinas de resposta Th2, compostas basicamente pela IL-4, IL-6 e IL-10, são responsáveis por bloquear a evolução do DM1. Através desses dados conclui-se que o entendimento dos aspectos imunológicos constitui a base para detecção e prevenção do DM1, sendo a disponibilidade de citocinas clonadas e purificadas uma nova perspectiva para terapias clínicas específicas para modular a resposta imune

Type 1 diabetes mellitus (DM1) is a chronic autoimmune disease, where as autoreactive T cells destroy as pancreatic beta cells leading to exogenous insulin dependence. For the development of DM1, interactions between immune system cells, mainly mediated by cytokines of T helper 1 (Th1) and T helper 2 (Th2) are required. The present study aimed to analyze the relationship of Th1 and Th2 response cytokines in the development of DM1, through a review of the literature, evaluating electronic scientific articles published between 2001 and 2016, as well as immunology books applied to the clinic. Several studies in the literature demonstrate that the cytokine secretion profile during the development of DM1 is of the Th1 pattern where we have as main constituent of IL-2 and IFN-γ. As Th2 response cytokines, composed mainly of IL-4, IL-6 and IL-10, they are responsible for blocking the progression of DM1. Through conclusive data, it is a point of view for the prevention of DM1. With the availability of cloned and purified cytokines a new perspective for specific clinical therapies to modulate the immune response