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Objective:To explore the possible correlation between serum detection of IL-7, IL-21, HBV-specific cytotoxic T lymphocytes (CTLs), HBV DNA, and the expression of CD127 on the T lymphocytes, and discuss the effect of IL-7 to cellular immune response in patients with chronic hepatitis B (CHB).Methods:Five hundred and sixty serum samples were collected from patients with CHB in Beijing Friendship Hospital from September 2017 to March 2020. The serum IL-7 and IL-21 were detected by enzyme-linked immunosorbent assay (ELISA), and HBV-specific CTLs and the expression of CD127 on the T lymphocytes were determined by flow cytometry. While HBV DNA were tested using quantitative real-time PCR (qRT-PCR). Subjects were divided into groups A, B, and C, according to the IL-7 levels (low: IL-7<20 pg/ml, medium: 20 pg/ml≤IL-7<30 pg/ml, and high: IL-7≥30 pg/ml).Results:The average concentration of serum IL-7 in patients with CHB was significantly lower than that of healthy controls ( P<0.01), and the difference among three groups was statistically significant ( P<0.01). Meanwhile, levels of IL-21, percentages of HBV-specific CTL, and the expression of CD127 on the CD8 + T lymphocytes showed an upward trend among groups, and there were significant differences among three groups ( P<0.01) with a positive correlation between each two variables ( P<0.01). However, HBV DNA showed a downward trend in group A, B and C, and the difference of the three groups were statistically significant ( P<0.01), which were negatively correlated with other variables ( P<0.01). Multiple linear regression analysis showed that HBV-specific CTL was an independent influencing factor for HBV DNA ( P<0.01), and IL-7, the expression of CD127 on the CD8 + T lymphocytes and IL-21 had an independent effect on HBV-specific CTL ( P<0.05). Conclusions:IL-7 could regulate HBV-specific immune response, and might be used as an effective cellular immune indicator to evaluate the cellular immune status of patients with chronic hepatitis B.
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Objective To detect the expression of follicuLar helper T cells (Tfh) and interleukin-21 (IL-21) in the peripheral blood of patients with hepatic echinococcosis and healthy controls, so as to explore the associations of Tfh and IL-21 expression with the progression of hepatic echinococcosis. Methods Fifty cases of hepatic echinococcosis and healthy controls were collected from Qinghai Provincial People's Hospital, respectively. Flow cytometry was used to detect the expression of Tfh cells in the peripheral blood of hepatic echinococcosis patients and healthy controls, and enzyme-linked immunosorbent assay (ELISA) was used to detect serum IL-21 expression in hepatic echinococcosis patients and healthy controls. The correlation between Tfh cell expression and serum IL-21 level was examined in the patients with hepatic echinococcosis. Results Flow cytometry detected a higher percentage of CD4+CXCR5+ T cells (18.49% ± 5.67% vs. 16.18% ± 4.04%, P < 0.05), CD4+CXCR5+PD-1+ T cells (4.94% ± 1.91% vs. 2.29% ± 0.79%, P < 0.05) and CD4+CXCR5+ICOS+PD-1+ T cells (30.93% ± 24.10% vs. 21.07% ± 14.25%, P < 0.05) in hepatic echinococcosis patients than in healthy controls, and no significant difference was seen in the percentage of CD4+CRCR5+ICOS+ T cells between the patients and controls (0.29% ± 0.32% vs. 0.25% ± 0.31%, P > 0.05) . The serum IL-21 level was significantly higher in the patients with hepatic echinococcosis than in healthy controls ([ 293.35 ± 2 03.65) pg/mL vs. (192.72 ± 70.09) pg/mL, P < 0.05]; however, there was no correlation between the Tfh cell expression and serum IL-21 level in patients with hepatic echinococcosis (P > 0.05). Conclusion The expression of peripheral blood Tfh cells and serum IL-21 is elevated in patients with hepatic echinococcosis, and Tfh cells and IL-21 may contribute to the progression of hepatic echinococcosis.
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Objective To detect the expression of follicuLar helper T cells (Tfh) and interleukin-21 (IL-21) in the peripheral blood of patients with hepatic echinococcosis and healthy controls, so as to explore the associations of Tfh and IL-21 expression with the progression of hepatic echinococcosis. Methods Fifty cases of hepatic echinococcosis and healthy controls were collected from Qinghai Provincial People's Hospital, respectively. Flow cytometry was used to detect the expression of Tfh cells in the peripheral blood of hepatic echinococcosis patients and healthy controls, and enzyme-linked immunosorbent assay (ELISA) was used to detect serum IL-21 expression in hepatic echinococcosis patients and healthy controls. The correlation between Tfh cell expression and serum IL-21 level was examined in the patients with hepatic echinococcosis. Results Flow cytometry detected a higher percentage of CD4+CXCR5+ T cells (18.49% ± 5.67% vs. 16.18% ± 4.04%, P < 0.05), CD4+CXCR5+PD-1+ T cells (4.94% ± 1.91% vs. 2.29% ± 0.79%, P < 0.05) and CD4+CXCR5+ICOS+PD-1+ T cells (30.93% ± 24.10% vs. 21.07% ± 14.25%, P < 0.05) in hepatic echinococcosis patients than in healthy controls, and no significant difference was seen in the percentage of CD4+CRCR5+ICOS+ T cells between the patients and controls (0.29% ± 0.32% vs. 0.25% ± 0.31%, P > 0.05) . The serum IL-21 level was significantly higher in the patients with hepatic echinococcosis than in healthy controls ([ 293.35 ± 2 03.65) pg/mL vs. (192.72 ± 70.09) pg/mL, P < 0.05]; however, there was no correlation between the Tfh cell expression and serum IL-21 level in patients with hepatic echinococcosis (P > 0.05). Conclusion The expression of peripheral blood Tfh cells and serum IL-21 is elevated in patients with hepatic echinococcosis, and Tfh cells and IL-21 may contribute to the progression of hepatic echinococcosis.
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@#[Abstract] Objective：To investigate whether the proliferation and cytotoxicity of NK-92MI cells can be improved by IL-7 and IL-21 genes modification, and determine the effects of this genetically modified NK-92MI cells on T cells from normal human peripheral blood. Methods：IL-7 and IL-21 gene fragments were constructed into electroporation vector by genetic engineering method, and NK92MI/IL-21 and NK-92MI/IL-7&21 cells were constructed by electroporation transfection. The in vitro proliferation and cytotoxicity of NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells were measured by cell count and flow cytometry assays. Then, normal human PBMCs were co-cultured with NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells in vitro respectively, and the phenotype change of T cells was measured by flow cytometry. In addition, the cytotoxicity between the activated T cells and three NK-92MI cell lines (NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells) as well as the cytotoxicity of the three NK-92MI cells on tumor cells after co-incubation with activated T cells were detected. Results: NK-92MI/IL-21 cell line (highly expressing IL-21) and NK-92MI/IL-7&21 cell line (highly expressing both IL-7 and IL-21) were successfully constructed. The toxicity of NK-92MI, NK-92MI/IL-21 and NK92MI/IL-7&21 cells on Jurkat and K562 cells showed no difference, while the proliferation of NK-92MI/IL-21 and NK-92MI/IL-7&21 cells was increased compared with NK-92MI cells. Furthermore, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells promoted the activation of T cells to a certain degree, and the activated T cells showed merely no cytotoxicity on NK-92MI, NK-92MI/IL-21 and NK-92MI/IL7&21 cells; Meanwhile, the activated T cells did not affect the cytotoxicity of the three NK cells (NK-92MI, NK-92MI/IL-21, and NK92MI/IL-7&21 cells) on K562 cells under their co-existence. Conclusion: The in vitro proliferation of NK-92MI/IL-21 and NK-92MI/ IL-7&21 cells were enhanced after gene modification, which could also stimulate and promote the activation of T cells from peripheral blood. The cytotoxicity assay showed that the activated T cells had no cytotoxicity on NK-92MI, NK-92MI/IL-21, and NK-92MI/IL-7& 21 cells. Meanwhile, the presence of the activated T cells did not affect the cytotoxicity of NK-92MI cells.
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@#[Abstract] Objective： ： To explore the impact of γ-chain (γc) family cytokines (IL-2, IL-7, IL-15, IL-21) on T cell phenotypes in ex vivo culture to provide experimental evidence for ex vivo cell preparation in adoptive immunotherapy. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood of healthy volunteers; nylon column sorting, CD3+ magnetic beads sorting, CD3- magnetic beads sorting and natural sedimentation were used to sort T cells from PBMCs. The purity, recovery rate and viability of T cells sorted by the above methods were compared. The CD3/CD28 magnetic beads-activated CD3+T cells were cultured inAIMV medium with IL-2 or mixed cytokines (IL-7, IL-15, IL-21). The expansion fold and phenotypes of T cells in ex vivo culture were detected by flow cytometry. Results： ： The purity of T cells sorted by CD3- magnetic beads sorting was significantly higher than that sorted by nylon column, CD3+ magnetic beads sorting and natural sedimentation ([94.06±1.07]% vs [86.74±1.06]%, [89.61±1.40]%, [88.48 ± 1.86]%, P<0.05); The recovery rate of T cells sorted by natural sedimentation was significantly higher than that by other three methods ([60.29±1.53]% vs [45.03±2.79]%, [20.15±3.41]%, [42.98±2.82]%, P<0.05). Comprehensively, the natural sedimentation method is the best option. The ex vivo expansion fold of T cells in IL-2 group was significantly higher than that in mixed group ([262.6±143.2] times vs [73.0±25.8] times, P<0.05). The proportions of early memory T cells, Tscm+Tscm-like and Tcmin the mixed group were significantly higher than those in the IL-2 group ([55.6±1.82]% vs [39.6±1.52]%, [16.6±1.82]% vs [9.8±1.30]%, [39.0±1.58]% vs [29.2±1.79]%; all P < 0.05). Conclusion： ： Natural sedimentation sorting has advantages of low cost, high recovery and purity. Mixed cytokines of IL-7, IL-15 and IL-21 are beneficial for production of early memory T cells. This study provides an experimental data of ex vivo T cell preparation for cancer adoptive immunotherapy.
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Objective To investigate the inhibitory effect and underlying mechanism of mesenchymal stem cell (MSC) derived from different sources on follicular helper T cell (Tfh cell). Methods Umbilical cord-derived MSC (UC MSC), bone marrow-derived MSC (BM MSC) and fat-derived MSC (Fat MSC) were co-cultured with peripheral blood mononuclear cell (PBMC) for 48 h. A control group was established. Flow cytometry was adopted to calculate the proportion of Tfh cells among the lymphocytes in four groups. The content of interleukin (IL)-21 in the supernatant was detected by enzyme-linked immune absorbent assay (ELISA) in four groups. BM MSC was co-cultured with PBMC, and supplemented with indoleamine 2,3-dioxygenase (IDO) inhibitor 1-methyl tryptophan (1-MT), IL-10 antibody, human leukocyte antigen (HLA)-G antibody in the 1-MT group, IL-10 inhibition group, HLA-G inhibition group and BM MSC group without addition of other substances. After 48 h culture, flow cytometry was used to detect the percentage of Tfh cells among lymphocytes. Results Flow cytometry demonstrated that compared with the control group, the proportion of Tfh cells in the BM MSC group was significantly decreased (P<0.05). Compared with the BM MSC group, the percentage of Tfh cells in the UC MSC and Fat MSC groups was significantly higher (both P<0.05). ELISA revealed that compared with the control group, the IL-21 content in the BM MSC group was significantly decreased (P<0.05). Compared with the BM MSC group, the IL-21 contents were considerably higher in the UC MSC and Fat MSC groups (both P<0.05). The analysis of underlying mechanism revealed that the proportions of Tfh cells in the 1-MT, IL-10 inhibition and the HLA-G inhibition groups were (1.75±0.07)%, (1.31±0.09)% and (1.50±0.03)%, respectively, which were significantly higher than (1.03±0.43)% in the BM MSC group (all P<0.05). Conclusions BM MSC exerts the highest inhibitory effect upon the differentiation of Tfh cell and IL-21. The mechanism underlying suppressing the differentiation of Tfh cells differentiation is probably correlated to promoting the secretion of IDO.
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Objective:To study the clinical significance of Th17 cell detection in patients with thymoma associated with myasthenia gra-vis(MG).Methods:From October 2015 to October 2017,78 patients with thymoma who treated in Cardio-Thoracic Surgery of Tianjin Medical University General Hospital and 15 normal controls were collected.Flow cytometry was used to detect the proportion of Th17 cells,and the levels of IL-17,IL-21 and AchR-Ab in peripheral blood were detected before operation.The correlation of Th17 with the corresponding cytokines was analyzed,and the independent risk factor was predicted for thymoma with myasthenia gravis.Results:The percentage of Th17 cells in the MG group was 10.33%±1.28%,compared with 7.33%±0.58% for the Con group and 7.67%±2.18% for the Tm group.These differences were statistically significant.The MG group was recorded as having(18.05 ± 7.23)pg/mL IL-17, (180.67±16.17)pg/ml IL-21,and(37.29±7.48)pmol/L AchR-Ab compared to(7.45±3.46)pg/mL,(84.33±5.13)pg/mL,and(15.23±3.32) pmol/L for the Con group and(8.22±1.67)pg/mL,(93.00±7.21)pg/mL,and(16.87±4.89)pmol/L for the Tm group for the same cyto-kines,respectively.These differences were also statistically significant(P<0.001).The percentage of Th17/CD4+T cells in the MG group was positively correlated with IL-17(r=0.830,P<0.001),IL-21(r=0.734,P<0.001),and AchR-Ab(r=0.809,P<0.001).IL-17,IL-21,and AchR-Ab are independent risk factors for predicting the combination of thymoma with MG.Conclusions:Th17 cells are involved in the development of thymoma with MG,which promote cell proliferation by secreting IL-21 and regulate MG growth by secreting IL-17.
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Objective:To investigate the expression and clinical significance of B7-H3 and IL-21 in serum of patients with HBV-related primary liver cancer.Methods: Gathering 121 cases of HBV-related patients,50 cases of primary hepatic carcinoma of them were considered as hepatic carcinoma group,71 cases of benign group including 12 cases with acute hepatitis,21 cases of chronic moderate to severe hepatitis,20 cases of compensatory phase cirrhosis,18 cases of decompensated cirrhosis and 20 cases of healthy persons in the same period as normal control.The content of serum B7-H3 and IL-21 were detected by ELISA.HBV DNA quantitative results were analyzed by Quantitative Real-time PCR.Results: The levels of B7-H3 and IL-21 in patients with primary hepatic carcinoma were (207.60±57.16)ng/ml and(2 357.28±805.01)pg/ml,respectively,which were significantly higher than those of the normal control subjects(P<0.001).Comparison with the normal control subjects,the content of B7-H3 and IL-21 in serum of patients with different clinical types in the benign group increased significantly(P<0.001).B7-H3 and IL-21 were positively associated with each other in serum of patients with HBV-related primary liver cancer.The expression of sB7-H3 was not significantly correlated with the degree of HBV DNA replication.The expression of IL-21 was correlated with HBV DNA replication in patients with HBV associated hepatocellular carcinoma,but was not significantly correlated with the degree of HBV DNA replication.Conclusion: HBV-related primary hepatic carcinoma express sB7-H3 and IL-21 with high level.The continuous high expression of sB7-H3 and IL-21 in the body may be related to the development and prognosis of the patients.
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Objective:To study the expression of IL-21/BCL-6/Blimp-1 in CE patients and discusse the mechanism of the pathogenesis of the echinococcosis.Methods:27 patients and 30 health persons were collected from the first affiliated hospital of Xinjiang medical university in the same period.IL-21 was detected by ELISA and the expression of IL-21/BCL-6 /Blimp-1 mRNA was detected by Real-time fluorescence quantitative PCR (qRT-PCR) in CE patients.At the same time,17 patients were followed up in the group of patients,and the expression of IL-21/BCL-6/Blimp-1 was detected before and after treatment.Results:(1) The results of PCR showed that the levels of IL-21/BCL-6 mRNA were significantly increased in the peripheral blood mononuclear cells of the CE patients compared with the healthy control group (P<0.05).The expression of IL-21/BCL-6 /Blimp-1 mRNA in patients before the treatment was higher than that of patients after treatment(P<0.05).(2)The level of IL-21 in peripheral blood of CE patients was sig-nificantly higher than that in the healthy control group and basically returned to normal after the cure (P<0.05).IL-21 was positively correlated with BCL-6(r=0.733, P<0.01).Conclusion:BCL-6 and Blimp-1 May promote the human immune system to resist parasitic infection in the course of the development of the disease.IL-21, BCL-6 and Blimp-1 are significantly reduced after effective treatment,indicating that these factors are involved in the immune mechanism of the development of the disease.
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Objective To optimize in vitro amplification of human γδ T cells with cytokines for tumor adoptive immunotherapy.Methods On the basis of the immobilized anti-TCR γδ antibody plus IL-2 system, other γ chain receptor family cytokines, including IL-7, IL-15 and IL-21, were tested to amplify human peripheral blood γδ T cells either alone or in diversity combination.The percentage of γδ T cells was measured by flow cytometry, and the proliferation efficiency of γδ T cells was calculated.The expression of proliferation-or cytotoxicity-related molecules on γδ T cells was examined by flow cytometry in order to explore the relevant mechanisms.The cytotoxicity of γδ T cells to Daudi cells was detected by lactate dehydrogenase.Results IL-15 alone but not IL-7 or IL-21 increases the γδ T cell purity, amplification efficiency and cytotoxicity to reach comparable levels to those of IL-2.IL-2 plus IL-15 up-regulates the expression of CD69 on γδ T cells and significantly increases their amplificationefficiency (P<0.05).IL-2 plus IL-21 enhanced the cytotoxicity of γδ T cells against Daudi cells by increasing the expression of granzyme A (P<0.001).The combination of IL-2, IL-15 and IL-21 significantly improves cytotoxicity of γδ T cells but reduces their amplification efficiency.In addition, when IL-21 was applied for a short time, it also enhanced the cytotoxicity of γδ T cells (P<0.05).Conclusions The combination of IL-2 and IL-15 as well as a short time addition of IL-21 is the best cytokine recipe to amplify human peripheral blood γδ T cells in vitro with immobilized anti-TCR γδ antibody, which can increase both the proliferation efficiency and the cytotoxicity to tumor cells of γδ T cells.
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AIM:To discuss the changes of IL-17, IL-21 in glucocorticoid therapy of active thyroid associated ophthalmopathy (TAO) and analyze the relation with results.METHODS:The 67 patients (134 eyes) of TAO in our hospital, were divided into activity group(32 patients 64 eyes) and atypical activity group(35 patients 70 eyes) according clinical activity score (CAS) grading standard, and 30 cases of healthy as control group at the same time.The activity group were treated by glucocorticoid therapy treatment, and proceed the CAS scores before and after treatment, measured the degree of exophthalmus and width of palpebral fissure.At the same time, compared the expression level of IL-17 and IL-21 in all groups, and analyzed the correlation between the IL-17 and IL-21 and CAS score.RESULTS:Compared with control group, expressions of IL-17 and IL-21 in TAO patients were significantly higher (P<0.05).The expressions of IL-17 and IL-21 in active period TAO patients were higher than atypical activity (P<0.05).After glucocorticoid treatment, the expressions of IL-17 in active period TAO patients decreased significantly(P<0.05), and it was significantly positive correlation with CAS score (before treatment:r=0.8847,P=0.042;after treatment:r=0.8886,P=0.0439) the expression of IL-21 in active period TAO patients was significantly positive correlation with CAS score (before treatment:r=0.8893, P=0.0435;after treatment:r=0.8876,P=0.045).CONCLUSION:IL-17 and IL-21 is closely related to the TAO disease activity, and glucocorticoids impact treatment by reducing IL-17 and IL-21 in activity TAO, IL-17 and IL-21 can be used as one of indexes of predicted curative effect and condition in patients with TAO.
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Objective:To explore the role of IL-21 in regulating the expression of peripheral blood Treg cells in the pathogenesis of Graves disease ( GD ) . Methods: Electrochemical luminescence detection was used to determine levels of thyroid function indexes and autoantibodies. Peripheral blood mononuclear cells (PBMCs) were extracted,then,cultured in the presence or absence of IL-21 in vitro. The level of IL-10 protein was measured by enzyme-linked immunosorbent assay ( ELISA) ,and expressions of Foxp3 and IL-10 mRNA were examined by Real-time PCR. Results: Compared to eGD and control group, there were significant differences in the levels of thyroid function indexes in GD group (P0. 05). Before IL-21 stimulation,compared to eGD and control groups,expressions of Foxp3 and IL-10 mRNA and level of IL-10 protein were significantly higher in GD group ( P0. 05). After IL-21 stimulation,expressions of Foxp3,IL-10 mRNA and IL-10 protein levels were decreased markedly in all three groups,among which GD group showed the greatest change(P<0. 05). Conclusion:IL-21 may inhibit the differentiation of Treg cells and the production of IL-10,therefore decreasing the number of Treg cells and the ability of suppressing effector T cells ,thus should be involved in the pathogenesis of GD.
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Objective:To examine the roles of follicular helper T(Tfh)cells,serum IL-21 and B cells in the pathogenesis of alcohol abuse non-traumatic osteonecrosis of the femoral head ( NONFH ) .Flow cytometry was used to measure the frequencies of peripheral blood inducible Tfh cells and B cells in alcohol abuse NONFH patients and healthy controls.The disease progression and the extent of femoral head collapse,the serum IL-21 were quantified.Methods: A significantly higher percentages of CD19+B cells(t=3.765,P=0.0005),CD86+CD19+B cells(t=5.506,P<0.0001),and CD95+CD19+B cells(t=4.152,P=0.0002) in patients than those in controls was found.The percentages of CD86+CD19+B cells were positively associated with the index of femoral head collapse in alcohol abuse NONFH(P<0.0001,r=0.536).Results: The frequencies of Tfh cells (t=7.611,P<0.0001),and IL-21+Tfh cells (t=5.281,P<0.0001) were higher than those in controls;The frequencies of Tfh cells were positively associated with the percentages of CD19+B cells(P=0.0002,r=0.455),IL-21+Tfh cells were positively associated with the percentages of CD86+CD19+B cells(P=0.0002,r=0.447).Conclusion: Tfh cells and B cells may participate in the pathogenesis of alcohol abuse NONFH,and increased CD86+CD19+B cells may be associated with the extent of femoral head collapse,the interaction of Tfh cells and B cells may have an im-portant role in pathogenesis of alcohol abuse NONFH.
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Objective:To analyse the role of Bcl-6/Blimp-1/IL-21 in pathogenesis of primary Sj?gren′s syndrome(PSS),the relationship between Bcl-6/Blimp-1/IL-21 expression and labial gland biopsy grading.Methods:Immunohistochemical(IHC) method was used to detect the expression of Bcl-6/Blimp-1 in salivary gland between 30 cases pations with PSS in disease group and 11 cases patients with mucous cyst,lower lip trauma in control group,and the serum IL-21 levels between disease group of 40 cases patients with PSS and 30 cases healthy volunteers were measured by enzyme-linked immunosorbent assay( ELISA) ,relations among 15 patients with IL-21 expression levels with labial gland biopsy grading.Results:The expression levels of Bcl-6/Blimp-1/IL-21 in disease group were higher than control group;in disease group,with pathological grades increased,the expression levels of Bcl-6/Blimp-1/IL-21 were also raised.Conclusion:①Bcl-6/Blimp-1/IL-21 may participate in the pathogenesis of PSS;Bcl-6/Blimp-1/IL-21 is associated with infiltration lymphocytes of salivary gland.
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Objective To determine the expression of interleukin-21 (IL-21) in patients infected with hepatitis B virus (HBV) and its association with HBV-DNA and ALT. Methods Clinical dates and blood specimen were collected from 25 unrelated healthy controls (HC) and 101 independent chronic HBV infected patients, including 25 patients in immune tolerant phase (IT), 25 in immune clearance phase (IC), 26 patients in inactive HBV carrier state (IA) and 25 patients in immune reactive phase (IR). Serum IL-21 levels were measured by Cytometric Bead Array (CBA). IL-21 mRNA and IL-21 receptor mRNA were measured by real-time PCR. Results Chronic HBV-infected patients had higher levels of serum IL-21 and IL-21 mRNA , with P <0.001 for both. In subgroup analysis, both serum IL-21 and IL-21 mRNA levels in IC, IR were higher than those in IT, IA and HC (all P < 0.001). Serum IL-21 level in IA was higher than that in HC and IT (P <0.001, P = 0.036). IL-21R mRNA levels were different between groups. Serum IL-21 level was associated with HBV-DNA (r = -0.472, P < 0.001), but not with ALT. Conclusion IL-21, up-regulated in chronic HBV infection, is associated with immune activity and may play a key role in HBV control.
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Objective To investigate the expression of T helper type 17 cells (Th17) and cell‐related cytokines ,including interleukin (IL)‐21 ,IL‐22 ,IL‐23 in the peripheral blood of different clinical stages of patients with acute hepatitis B (AHB) .Methods Ten cases of AHB patients were enrolled .The frequency of Th17 cells in the three clinical stages (i .e .acute phase ,convalescent phase and resolved phase) were detected by flow cytometry . IL‐21 , IL‐22 and IL‐23 were measured by enzyme‐linked immunosorbent assay (ELISA) .Control group was composed of ten healthy subjects .The comparison between the two groups was done by t test and the differences among multiple groups were compared by one way ANOVA .Pearson correlation analysis was used for correlation analysis .Results The frequency of Th17 in healthy controls was (0 .68 ± 0 .29)% ,while those in acute phase ,convalescent phase and resolved phase of AHB patients were (18 .22 ± 4 .13)% , (3 .14 ± 1 .90 )% and (3 .31 ± 0 .95 )% , The differences between the two groups were significant (t= 13 .405 ,4 .047 and 8 .342 , respectively ;all P< 0 .01) .The levels of IL‐21 ,IL‐22 and IL‐23 in healthy controls were (42 .00 ± 6 .95) ,(315 .89 ± 96 .16) and (11 .95 ± 6 .95) ng/L ,respectively .Those in acute phase of AHB patients were (575 .39 ± 47 .01) ,(648 .44 ± 47 .12) and (38 .29 ± 4 .68) ng/L ,respectively ,those in convalescent phase were (366 .50 ± 33 .74) ,(405 .04 ± 47 .12) and (25 .10 ± 4 .69) ng/L ,respectively ,while those in resolved phase of AHB patients were (46 .62 ± 8 .28) ,(365 .94 ± 45 .62) and (15 .29 ± 4 .69) ng/L , respectively .Compared with healthy controls ,t values of the levels of IL‐21 in three different phases of AHB patients were 35 .497 ,29 .792 and 1 .354 with P value of <0 .01 ,<0 .01 and 0 .193 ,respectively ;those of IL‐22 were 9 .820 ,2 .632 and 1 .487 with P value of < 0 .01 ,0 .021 and 0 .161 ,respectively ;those of IL‐23 were 9 .944 ,4 .961 and 1 .260 with P values of <0 .01 ,<0 .01 and 0 .226 ,respectively . After comparison of IL‐21 ,IL‐22 and IL‐23 among three different phase of AHB ,F values were 622 .784 , 107 .772 and 60 .743 with all P values less than 0 .01 ,respectively .The levels of IL‐21 ,IL‐22 and IL‐23 were all positively correlated with the serum ALT level in acute phase (r= 0 .655 ,0 .666 and 0 .673 , respectively ;all P<0 .05) .Correlation analysis demonstrated that the frequency of Th17 was positively correlated with the levels of IL‐21 , IL‐22 and IL‐23 in acute phase ( r= 0 .879 ,0 .866 and 0 .879 , respectively ;all P<0 .01) .The frequency of Th17 was positively correlated with the level of IL‐21 in the resolved phase . No correlations between the remaining groups were confirmed . Conclusion The expressions of Th17 and cell‐related cytokines ,including IL‐21 ,IL‐22 and IL‐23 decline with the recovery of A HB .
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Objective To detect the level of IL-10 and IL-21 in serum of patients with psoriasis and explore its significance of judge the extent of serious condition and treatment of psoriasis.Methods According to clinical dermatology edited by Zhao Bian diagnostic criteria,selected 65 cases of patients with psoriasis during the period from March to September in 2014 ad-mitted to Department of Dermatology of out-patient and hospitalization.25 cases were progressive stage (group A),20 pa-tients with stable stage (group B),20 patients were in recovery period (group C),and 25 cases of control group (group D). With ELISA method detect the expression level of IL-10 and IL-21 in serum of each group respectively.PASI score will take to all psoriasis patients meanwhile.Results The expressionlevels of IL-21 in serum of A (127.59 ± 16.09 pg/ml),B (105.74±21.08 pg/ml)two groups were significantly higher than that in D group (85.46±14.25 pg/ml),and the differ-ence were statistically significant (t=5.174,4.863,both P <0.01),while the expression levels of IL-10 in A (10.64±3.23 pg/ml)and B (12.27±2.18 pg/ml)were both lower than D group (20.29±2.51 pg/ml),and the difference were statisti-cally significant (t=2.031,2.027,both P <0.05).The expression of IL-21 (94.03±8.90 pg/ml)in C group was higher than that in D group,and the difference was statistically significant (t=2.033,P <0.05).The level of IL-21 in A group was higher than that in group C,and the difference was statistically significant (t=2.352,P <0.05),while the expression level of IL-10 was lower than that in C group (19.69±1.54 pg/ml),and the difference was statistically significant (t=2.071,P<0.05).The expression level of IL-21 in the serum of patients with psoriasis and PASI score was positively correlated (r=0.508,P =0.027),while the expression level of IL-10 and PASI score was negatively correlated (r=-0.413,P =0.039). Conclusion The progression of psoriasis is related to the increasion of IL-21 or the decreasion of IL-10.The detection of IL-21 and IL-10 was beneficial to judge the severity and efficacy evaluation of psoriasis.
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Background: This study was conducted in order to investigate the association between IL-21 (rs2055979 G/T and rs13143866 A/G) and Foxp3 (rs2232365 A/G and rs3761548 A/C) gene polymorphisms and recurrent pregnancy loss (RPL) in a group of Palestinian women residing in Gaza strip. Methods: A retrospective case-control study was carried out during the period (January to June, 2014). A total of 200 females, 100 RPL patients and 100 controls women without previous history of RPL, aged 20–35 years were included in the study. IL-21 rs13143866 A/G and Foxp3 rs2232365 A/G polymorphisms were analyzed by allele-specific PCR whereas, IL-21 rs2055979 G/T and Foxp3 rs3761548 A/C polymorphisms were investigated by PCR-RFLP. Results: No statistically significant difference existed between RPL cases and controls in terms of the allelic and genotypic distribution of IL-21 rs2055979 or rs13143866. On the other hand, the two Foxp3 gene polymorphisms showed statistically significant association with RPL: the rs2232365 A/G allele G and its homozygous genotype G/G and the rs3761548 A/C allele A and its A/A genotype were significantly higher in RPL group. Conclusions: Foxp3 rs2232365 A/G and rs3761548 A/C polymorphisms represent risk factors for RPL in Gaza strip women. IL-21 polymorphisms rs2055979 G/T and rs13143866 A/G however, do not pose tangible risk for RPL in our population. Study results should be confirmed on a larger sample.
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Objective:To express recombinant protein mIL-21-hIgGFc in 293E cells,and investigate its effect on CD8+T cell.Methods:Total RNA was extracted from the mouse spleen cells ,and then IL-21 gene was amplified by RT-PCR and inserted into expression vector PTT3-hIgGFc.PTT3-mIL-21-hIgGFc were transfected into 293E cells by calcium phosphate method.The supernatants were collected at 48 hours and 72 hours and concentrated by MOLLIPORE Labscale TM TFF system ( 5 kD membrane ).The mIL-21-hIgGFc fusion protein was purified with HiTrap TM Protein G column.The protein was quantified by SDS-PAGE and ELISA.The biological activity of the protein was determined by detecting the change of the phenotypes of CD 8+ T cells treated with the protein.Results: The constructed recombinant plasmid PTT 3-mIL-21-Fc was confirmed by sequencing.PTT3-mIL-21-Fc was transfected into 293E cells,mIL-21-Fc protein in culture supernatant was collected after 48 hours and 72 hours.The protein in cell su-pernatant reached a concentration of 787 ng/ml which was determined by ELISA.The protein was purified by Protein G chromatography column.P1A-specific T cells were treated with mIL-21-hIgGFc, and found that the CD44low CD62Lhi CD8+ population increased compared to the control.Conclusion:We built PTT3-mIL-21-hFc recombinant plasmid, expressed mIL21-hFc fusion protein in 293E cells,and purified by Protein G column.By treating mIL-21-hFc ,the antigen-primed CD8+T cells prefer to differentiate into CD44low CD62Lhi CD8+T cells which had been reported as a memory stem phenotype .This protein may be used to improve the effectness of adoptive T cell cancer therapy.
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Objective To investigate the possible mechanism of hypogammaglobuinemia caused by ketongenic diet (KD).Methods Thirty-six children with intractable epilepsy (IP) and seventeen age-matched healthy children were recruited in this study.The percentages of B cells at various stages of devel-opment and follicular helper T ( Tfh) cells were detected by flow cytometry.The plasma concentrations of IL-21 were determined by ELISA.Real-time quantitative PCR was performed to detect the expression of mam-malian target of rapamycin ( mTOR) , Blimp-1, Bcl-6 and IL-21 at mRNA level in CD4+T cells.Results mTOR at mRNA level was significantly down-regulated after KD treatment (P<0.05).The numbers of Tfh cells were positively correlated with the transcriptional level of mTOR (r=-0.691, P<0.05).Conclusion KD treatment might down-regulate Tfh and B cells through suppressing the expression of mTOR at mRNA level, suggesting a possible mechanism of hypogammaglobuinemia induced by KD treatment.