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Objective:To explore the changes of T follicular regulatory (T FR) cells/T follicular helper (T FH) cells and their related cytokines in peripheral blood of children with dust mite allergic asthma, and their clinical significance. Methods:A total of 25 children with acute dust mite allergic asthma (the asthma group) in Affiliated Hospital of Jiangnan University from January to December 2021 and 16 age- and sex-matched healthy volunteers (the healthy control group) at the same time were enrolled in the retrospective study.The percentages of peripheral T FR cells and T FH cells of the 2 groups were measured by flow cytometry.The plasma levels of cytokines[interleukin (IL)-10, IL-21] of the 2 groups were assessed by the flow cytometric microsphere-based array technology.The specific IgE (sIgE) levels of dust mites in 2 groups were detected by fluorescence enzyme immunoassay.The percentage of eosinophils in peripheral blood detected by blood cell analyzer.Data between groups were compared by t-test, and the correlation among indicators was analyzed by Spearman rank correlation analysis. Results:The asthma group had evident T FR cells/T FH cells immune imbalance.Compared with the healthy control group, the asthma group had a significantly lower T FR cells level[(0.11±0.03)% vs.(0.13±0.03)%], a significantly higher T FH cells level[(5.07±1.75)% vs.(3.80 ± 1.60)%], and a significantly lower ratio of T FR cells /T FH cells(0.02±0.01 vs.0.05±0.03) ( t=2.29, 2.30, 3.71; all P<0.05). Compared with the healthy control group, the asthma group had a significantly higher IL-21 level[(547.85±195.13) ng/L vs.(404.94±110.41) ng/L], and a significantly lower IL-10 level[(10.18±3.49) ng/L vs.(14.79±5.65) ng/L] ( t=2.60, 3.15; all P<0.05). The ratio of T FR cells/T FH cells in asthma group was negatively correlated with sIgE ( r=-0.444 2, P=0.026 1), but not related to the eosinophil percentage ( r=-0.135 2, P=0.519 3). Conclusions:Children with dust mite allergic asthma suffer from T FR cells/T FH cells subset imbalance.The imbalanced T FR cells, T FH cells and their related cytokines IL-10 and IL-21 may play a role in regulating the production of asthma sIgE.
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Objective To investigate the changes of expression levels of interleukin?17(IL?17) and interleukin?21( IL?21) secreted by helper T cells 17 before and after treatment of three immune?related hematological diseases,and to explore the clinical significance??Methods Sixty patients with IRH admitted to the Department of Hematology,the First Affiliated Hospital of Hebei North University from May 2017 to October 2018 were selected as subjects??They were divided into aplastic anemia group (24 cases),immune thrombocytopenia group (20 cases) and autoimmune hemolytic anemia group (16 cases)??Another 60 healthy volunteers who had physical examination in our hospital at the same time were selected as the control group??In the 3 IRH groups,patients were treated with glucocorticoids or immunologic agents,and the control group was given vitamin C??At the time of initial diagnosis,2 weeks and 3 months after IRH treatment,all the patients in the 4 groups were examined for blood?related indicators,and the therapeutic effects of different stages and the plasma levels of IL?17 and IL?21 in the peripheral blood of the 4 groups before and after treatment were compared??Results At the initial diagnosis,the plasma IL?17 levels in the aplastic anemia group,immune thrombocytopenia group and autoimmune hemolytic anemia group were ((196??52± 17??46), (185??69± 18??19),(126??13 ± 11??22) ng／L),respectively,which were higher than those in control group ((72??36± 10??21) ng／L), the differences were statistically significant ( all P＜0??05)??The plasma IL?21 levels in the aplastic anemia group,immune thrombocytopenia group and autoimmune hemolytic anemia group were ((136??82±20??16),(145??92±22??18),(119??66±12??69)) ng／L,respectively,which were higher than those in control group (( 84??01 ± 9??87) ng／L), and the differences were statistically significant ( all P＜0??05)??After 2 weekly treatment,the levels of IL?17 and IL?21 in the 3 groups were significantly lower than those at the first visit (all P＜0??05)??After 3 months of treatment,the levels of IL?17 and IL?21 in the 3 groups were also significantly lower than those at the first diagnosis ( IL?17: aplastic anemia group ( 84??69 ±12??15) ng／L,immune thrombocytopenia group (90??56±11??64) ng／L,autoimmune hemolytic anemia group (62??83±5??68) ng／L;IL?21: aplastic anemia group ( 96??28± 8??84) ng／L,immunological thrombocytopenia group (103??21±10??62) ng／L,autoimmune hemolytic anemia group (78??64±9??68) g／L),and the difference were statistically significant ( all P＜0??05)??There was no significant change in the control group ( 83??84 ±9??95) ng／L(P＞0??05)??After 3 months of treatment,the total effective rates of treatment in the three groups (the aplastic anemia group 83??33%( 20／24), immunological thrombocytopenia group90??00%( 18／20), autoimmune hemolytic anemia group 75??00%( 12／16)) were higher than 2 weekly treatment ( aplastic anemia group 41??67%( 10／24 ), immunological thrombocytopenia group 40??00%( 8／20 ), autoimmune hemolytic anemia group25??00%(4／16)),and the differences were statistically significant( all P ＜0??05)??Conclusion The changes of plasma IL?17 and IL?21 levels are helpful to indicate the occurrence and progression of immune?related hematopathy,to find therapeutic targets and to improve prognosis,which has important clinical significance in the clinical diagnosis,prognosis and treatment of different types of IRH??
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Objective To evaluate the therapeutic effect of baicalin on lupus nephritis in a lupusprone mouse model,and to investigate its regulatory role in the differentiation of follicular helper T (Tfh)cells.Methods Eight 12-week-old female MRL/lpr lupus-prone mice were randomly and equally divided into two groups by a random number table i.e.,baicalin group and control group intraperitoneally injected with 200 mg/kg baicalin sodium and chloride physiological solution,respectively,once every day for 4 weeks.After the end of treatment,Coomassie brilliant blue staining was performed to detect the level of 24-hour urine protein.Then,the mice were sacrificed,and the spleens were resected and weighed.Mononuclear cells were isolated from these spleens,and flow cytometry was conducted to determine the proportion of Tfh cells.Additionally,the kidneys were resected and subjected to hematoxylin and eosin (HE) staining for the evaluation of kidney impairment.Moreover,some other mononuclear cells were isolated from the spleens of the lupus-prone mice in the control group,and magnetic activated cell sorting (MACS) was performed to isolate naive CD4+ T cells,which were divided into 3 groups:blank control group receiving no treatment,induction group treated with 10 μg/L anti-interleukin (IL)-21 and anti-IL-6 antibodies and 3 μg/L anti-CD3 and anti-CD28 antibodies for 5 days,and intervention group additionally treated with 40 μmol/L baicalin for 5 days besides the above treatment.Then,50 μg/L phorbol ester,750 μg/L ionomycin and 20 mg/L brefeldin A were used to stimulate some cultured naive CD4+ T cells in the above groups.Flow cytometry was conducted to determine the proportion of CD4+CXCR5+PD-1 + cells and CD4+IL-21+ cells.Statistical analysis was carried out with SPSS20.0 software by using one-way analysis of variance (ANOVA) and student t test for the comparison of quantitative data between groups.Results The baicalin treatment could effectively improve the kidney impairment in the lupus-prone mice.Compared with the control group,the baicalin group showed significantly decreased 24-hour urine protein level ([1 416 ± 171] vs.[2 623 ± 278] μg/24 h,P =0.022),and significantly decreased proportion of Tfh cells in the spleen (12.6％ ± 2.3％ vs.40.2％ + 1.1％,P =0.005).In vitro baicalin could further inhibit the differentiation of Tfh cells.Compared with the induction group,the intervention group showed significantly decreased proportion of CD4+CXCR5+PD-1+ Tfh cells (13.3％ ± 0.8％ vs.17.6％ ± 0.9％,P =0.04) and CD4+IL-21+ cells (1.0％ ± 0.4％ vs.2.7％ ± 0.2％,P ＜ 0.01).Conclusion Baicalin can effectively ameliorate lupus nephritis,which may be associated with the inhibition of Tfh cell differentiation.
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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 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 investigate the correlations of the serum level of interleukin-21 (IL-21) with anti-BP 180 antibodies,peripheral eosinophil level,bullous pemphigoid disease area index (BPDAI)in patients with bullous pemphigoid (BP).Methods A total of 31 patients with bullous pemphigoid and 31 healthy controls were enrolled into this study.Blood samples were collected from all the subjects,and enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum level of IL-21 and serum titer of anti-BP180 antibodies.BPDAI was assessed among 13 patients,and the correlation of the IL-21 level with anti-BP180 antibody titers,eosinophil count and BPDAI were analyzed.Results No significant difference in the serum level of IL-21 was observed between the patients (M [P25-P75]:255.28 [156.19-361.59] ng/L) and healthy controls (150.47 [72.4-268.73] ng/L,P =0.14).There was no correlation between the serum level of IL-21 and serum titer of anti-BP180 antibodies (82.98 [26.82-132.92] U/ml,rs =0.18,P =0.332).Among the 27 BP patients who underwent blood cell count,no correlation was found between the serum levels of IL-21 and whole-blood eosinophil counts (rs =0.104,P =0.606).Among the 13 BP patients who underwent assessment of BPDAI,there was no correlation between the serum levels of IL-21 and BPDAI scores (rs =0.233,P =0.464).Conclusion The IL-21 level is uncorrelated with anti-BP 180 antibody level,peripheral eosinophil level or BPDAI in patients with BP.
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Objective To investigate the secretory capacity and apoptosis of interleukin ( IL)-21 induced normal B cells by co-culture with serum from patients with systemic lupus erythematosus (SLE). Methods Serum from twenty new-onset SLE patients and 20 healthy donors were collected .CD1+9 B cells from the normal controls were co-cultured with serum from SLE patients in the presence or absence of IL-21-R-FC(4 μg/ml).Supernatant IgG and IgM concentration were measured by immunoturbidimetric assay on day 5.Supernatant anti-dsDNA level was determined by ELISA .The percentage of apoptotic cells was detected by flow cytometer.Results IgG,IgM and anti-dsDNA levels in normal B cells with SLE serum were significantly higher than those in the serum of SLE patients alone [ ( 5.84 ±1.79 ) g/L vs ( 4.25 ± 1.48)g/L,P=0.000;(0.46 ±0.21)g/L vs (0.43 ±0.21)g/L,P=0.003;(127.76 ±70.24)IU/ml vs (115.15 ±63.88) IU/ml,P=0.014 respectively].However, no significant differences were found in the group of normal B cells with non-homologous serum from normal controls (P>0.05).Supernatant IgG, IgM and anti-dsDNA levels in normal B cells with SLE serum significantly decreased while IL-21R-fusion protein was added [(5.26 ±1.62)g/L vs (5.84 ±1.79)g/L, P=0.006;(0.42 ±0.20)g/L vs (0.46 ±0.21)g/L, P=0.002;( 118.00 ±69.62 ) IU/ml vs ( 127.76 ±70.24 ) IU/ml, P =0.012 respectively ] .The apoptotic rate of B cells with SLE serum was significantly higher than that with normal serum [ ( 47.88 ± 12.65)%vs (38.86 ±10.32)%,P =0.004].But adding IL-21R-fusion conversed the apoptotic rates [(42.08 ±12.52)%vs (47.88 ±12.65)%,P=0.001].Conclusions SLE serum could induce normal B cells to form immunoglobulin secreting cells and producing autoantibodies , or apoptosis in pathological conditions.IL-21 might be considered as a potential therapeutic target of SLE .
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Objective To explore the relationship between viral load and expression of CD 226 on the surface of peripheral blood T follicular helper cells (Tfh) in patients with chronic hepatitis C (CHC) . Methods One hundred and thirty-five CHC patients hospitalized at Wuxi Fifth People′s Hospital from March 2015 to April 2017 were collected ,and another 30 healthy blood donors were set as healthy control group .CHC patients were divided into two groups based on hepatitis C virus (HCV) RNA level ,with 49 cases (36 .3％ ) in low viral load group and 86 cases (63 .7％ ) in high viral load group .Expression of CD226 on the surface of peripheral blood Tfh cells , Tfh cells ,interleukin (IL )-21 and HCV specific cytotoxic lymphocyte (CTL) levels of two patient groups were compared .Categorical data were compared with chi-square test and normally distribute continuous data were compared with t test .Correlations between different factors were analyzed by Pearson correlation analysis .Results Expression of CD226 on the surface of peripheral blood Tfh cells in 135 cases of CHC patients was (77 .69 ± 5 .42)％ ,which was lower than that of healthy control ([90 .06 ± 5 .83]％ ) ,and the difference between the two groups was significant (t= 7 .541 , P < 0 .01) .The CD226 expression on the peripheral blood Tfh cells in low viral load group was (88 .75 ± 6 .68)％ ,which was higher than that of high viral load group ([69 .23 ± 5 .86]％ ) ,and the difference between the two groups was significant (t = 19 .232 , P< 0 .01) .The viral load was negatively correlated with Tfh cell surface CD 226 expression (r = - 0 .705 , P < 0 .01) .The peripheral blood Tfh cell level in 135 CHC patients was higher than that of healthy control ,and the difference between the two groups was significant (t= 13 .878 , P< 0 .01) .The peripheral blood Tfh cell level in low viral load group was higher than that in high viral load group ,and the difference between the two groups was significant (t= 26 .993 , P< 0 .01) .The IL-21 level of 135 CHC patients was lower than that of healthy control ([70 .35 ± 1 .6]％ ) ,and the difference between the two groups was significant (t=18 .322 , P< 0 .01) .The IL-21 level in peripheral blood of low viral load group was higher than that of high viral load group ,and the difference between the two groups was significant (t= 84 .54 , P< 0 .01) . HCV specific CTL level in peripheral blood of low viral load group was higher than that of high viral load group ,and the difference between the two groups was significant (t = 29 .869 , P< 0 .01) .The viral load was negatively correlated with levels of HCV specific CTL (r= -0 .734 ,P< 0 .01) .Conclusions In patients with chronic hepatitis C ,different levels of viral load can result in different levels of CD 226 expression on the peripheral blood Tfh cells .Patients with low viral load has high CD226 expression on Tfh cell surface , resulting in rise of Tfh cell level ,IL-21 level and HCV specific CTL level .
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Objective:To research the correlation of serum levels of IL-21,TGF-β1,TNF-α and IgA1 in children with allergic purpura and the purpura nephritis.Methods:57 cases with allergic purpura who were treated in our hospital from November 2015 to May 2016 were selected and 29 cases were diagnosed with general allergic purpura,and another 28 cases were diagnosed with purpuric nephritis.30 healthy children were selected as the control group.Then the serum levels of IL-21,TGF-β1,TNF-α,IgAl,immunoglobulin A (IgA),C3 and C4 between the three groups were observed and compared.Results:The serum levels of IL-21,TGF-β1,TNF-α,IgA1 and IgA in the purpuric nephritis group were higher than those of the control group and the general allergic purpura group,and the differences were statistically significant (P＜0.05);The serum levels of IL-21,TGF-β1,TNF-α,IgA1 and IgA in the general allergic purpura group were higher than those of the control group,and the differences were statistically significant (P＜0.05);There was no statistically significant difference about the C3 and C4 in the three groups (P＞0.05).Conclusion:The serum levels of IL-21,TGF-β1,TNF-α and IgA1 may be involved in the development ofhenoch-schonlein purpura and purpura nephritis.
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The etiology and pathogenesis of nephrotic syndrome is not fully clear.Most pepole believe that it is associated with the disorder of immune function.As immunoregulatory factors,interleukins(IL) and interferon(IFN) are closely related with the onset of nephrotic syndrome.In this paper,we review the role of IL-21 and INF-γin pathogenesis of nephrotic syndrome.
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Objective To investigate the roles of SENP1 in regulation of biological characteristics of NK cells.Methods Lentivirus-mediated-Senp1-small-hairpinRNA (shRNA) transduction was applied to NK92 cells.The expression of SENP1 in NK92 cells was determined by real-time PCR and Western blot.The proliferation of NK92 cells was detected by CCK-8 assay.The apoptosis of NK92 cells was determined by Annexin Ⅴ and PI labeling.The cytotoxicity of NK92 cells against K562 cells was evaluated by luciferase reporter assay.Results Treatment of NK92 cells with IL-21 resulted in SENP1 upregulation.Lentivirus mediated SENP1 knockdown reduced proliferation and increased apoptosis in NK-92 cells,but SENP1 inhibition had slight impact on the cytotoxic ability of NK92 cells to kill K562 cells.Conclusion SENP1 mediates the regulatory effect of IL-21 on the proliferation and survival of NK92 cells.
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Objective To explore the effect of CD40 small interfering RNA(siRNA)on the expressions of pe-ripheral blood interleukin(IL)-21 and IL -35 in rats with experimental autoimmune myocarditis (EAM)and its sig-nificance.Methods Twenty 6 -8 week male Lewis rats were divided into normal group,EAMgroup,CD40 siRNA group and siRNA group by using random number table,with 5 rats in each group.The normal rats were induced with phos-phate buffer saline in double foot pads on day 0 and day 7,while the rest 3 groups were induced with cardiac myosin protein to establish EAMmodels.The rats in CD40 siRNA group and siRNA group were respectively injected with CD40 siRNA and siRNA slow virus expression vector through the tail vein of rats on day 7.The rats were executed on 21 day after echocardiogram examination was made.The histopathologic changes were observed by using light microscope and the myocardial histopathology scores were calculated.Enzyme -linked immunosorbent assay was used to determine the levels of IL -21 and IL -35 in peripheral blood.Results (1)Except the normal group,the total incidence rate of rats of each group was 100%,and there was no rat death.(2)Compared with EAM group,the heart mass/body ratio and myocardial histopathology scores were lower in CD40 siRNA group,and the differences were significant (3.13 ±0.21 vs 3.80 ±0.29,2.22 ±0.43 vs 3.32 ±0.51,F =0.332,0.456,all P <0.05).(3)The echocardiogram showed that there was only 1 rat in EAM group with massive pericardial effusion,and there was no pericardial effusion in CD40 siRNA group.EAMgroup,CD40 siRNA group and siRNA group displayed hypertrophy of the ventricular septum and left ventricular wall,narrow heart cavity and weakening of ventricular wall motion.The left ventricular shortening rate in CD40 siRNA group was significantly higher than that in the EAMgroup[(63.34 ±11.06)% vs (38.56 ±6.98)%,F =16.080,P <0.05].(4)The peripheral blood level of IL -21 in CD40 siRNA group was lower than that in EAM group [(141.19 ±17.46)ng/L vs (157.81 ±17.58)ng/L,F =57.008,P <0.05],while its level of IL -35 was signifi-cantly higher than that in the EAMgroup [(195.96 ±18.26)ng/L vs (174.78 ±13.91 )ng/L,F =31.727,P <0.05].(5)The level of IL -21 in peripheral blood was positively correlated with myocardial histopathology scores in EAM group (r = 0.69,P < 0.05 ),but IL -35 was negatively correlated with myocardial histopathology scores (r =-0.64,P <0.05).Conclusions CD40 siRNA might relieve the myocardial inflammation and reduce the myocar-dial injury of EAMrats.The levels of IL -21 and IL -35 can partly reflect the degree of myocardial injury.The mecha-nism may be related to down -regulating the expression IL -21 and up -regulating the expression of IL -35.
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Objective To investigate the changes of follicular helper T cells (Tfh cells) and Tfh cells associated molecules in the peripheral blood (PB) of patients with malignant lymphoid diseases (MLD) dynamically, and explore their roles on pathogenesis of the diseases. Methods Fifty-five patients with MLD were enrolled in this study,including 9 patients with acute lymphocyte leukemia (ALL), 30 patients with non-Hodgkin lymophoma (NHL) and 16 patients with multiple myeloma (MM), and 10 healthy controls (NC) of similar age were also enrolled. The percentage of CD4+CXCR5+cells (Tfh cells) and expression of ICOS+, PD1+among the T cells were detected by flow cytometry (FCM), while the levels of interleukin 21 (IL-21) in plasma were detected by ELISA tests. Results The percentage of Tfh cells and expression of ICOS and/or PD-1 in PB of all untreated patients were significantly higher than those of NC (all P 0.05), and apparently lower than those who achieved PR (P 0.05), and much higher than NC (P< 0.01). The concentration of IL-21 in patients were much higher than that in NC [(326.56±32.44) pg/ml] (P<0.01), and MM group
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Objective To explore the impact of ketogenic diet (KD) on follicular helper T cells(TFH) in children with intractable epilepsy.Methods Thirty-three cases with intractable epilepsy were selected between Jul.2013 and Jan.2014 in Shenzhen Children's Hospital,19 boys and 14 girls; average age was 39.6 months,and seventeen age-matched healthy children who took a physical examination in the same hospital were assigned as the healthy control group.Blood samples were collected from the children with refractory epilepsy before and after 1 week of KD treatment.The proportions of the various stages of B cells and TFH cells were detected by flow cytometry.The plasma concentration of interleukin-21 (IL-21) was determined by enzyme-linked immunosorbent assay(ELISA),and realtime quantitative PCR(RT-PCR) was performed to detect the levels of peroxisome proliferator-activated receptor gamma (PPAR-γ),B-lymphocyte-induced maturation protein-1 (Blimp-1),B-cell lymphoma 6 (Bcl6) and IL-21 mRNA expression in CD4 + T cells.Results (1) The number of TFH cells in children with intractable epilepsy [(3.57 ± 0.58) ％] was remarkably decreased after KD treatment(P ＜ 0.01),while there were no difference between after KD treatment and healthy control group[(4.93 ±0.70)％ vs (5.03 ±0.63)％,P ＞0.05].(2) The levels of transcription factor Bcl6 expression after treatment were significantly decreased,while inhibitory factor Blimp-1 expression increased (P ＜ 0.05).(3)The plasma concentration of IL-21 had a trend to decrease (P ＞ 0.05),while there were no difference before and after KD treatment,and levels of IL-21 mRNA expressions in CD4 +T cells were significantly decreased after the treatment (8.28 × 10-3 ± 1.19 × 10-3 vs 1.72 × 10-2 ± 0.81 × 10-2,t =3.08,P ＜ 0.05).(4) There was no significant difference in CD27-IgD + B cells before and after KD treatment (P ＞ 0.05),CD27 + IgD + B cell and CD27-IgD-B cells had a trend to decrease after KD treatment(P ＞0.05),and CD27 + IgD-B cells and CD27 + IgD-CD38 high plasma cells were significantly decreased after KD treatment (P ＜ 0.05).(5) The number of TFH cells were correlated positively with the number of CD27 + IgD-B cells and CD27 + IgD-CD3g high plasma cells (r =0.785,0.745,P ＜ 0.05).(6) The levels of PPAR-γmRNA in CD4 + T cells expression were significantly up-regulated after KD treatment (3.49 × 10-3 ± 1.10 × 10-3 vs 2.28 ± 10-3 ± 1.30 × 10-3,t =3.41,P ＜0.05),and the number of TFH cells and PPAR-γgene expression was correlated negatively (r =-0.619,P ＜ 0.05).Conclusions KD might down regulate TFH cell number and function through inducing PPAR-γexpression and could inhibit B cell differentiation,which might be one of the factors for hypogammaglobuinemia by KD treatment.
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Objective To observe recombinant plasmids were constructed with the macrophage colony-stimulating factor ( GM-SCF) , interleukin -21 (IL-21) and retinoic acid early transcription factor-1 (Rae-1), and observe the inhibitory effects in subcutaneous liver cancer model in mice with the recombinant plasmids.Methods The recombinant plasmids of GM-SCF, IL-21 and Rae-1 were constructed with RT-PCR method, mouse model was constructed, the model mice were randomly divided into six groups including control, IRES/GFP, IRES/IL21, IRES/GM-SCF, IRES/GM-SCF-IL21 and IRES/combination with 10 mice included in each group, each groups (15 mice) were treated with the corresponding gene therapy.The survival rate were observed after 60 days.The blood levels of interferon -γ(IFN-γ) and interleukin -2 (IL-2) were detected in each group.Results The pGM-CSF-GFP-IRES-Rae-1-IL-21 has been successfully constructed.All mice had demised 14 and 16 days after treatment in the control and IRES/GFP groups, respectively.There were 2, 1, 11 mice remaining after 60 days of treatment in the IRES/GM-SCF, IRES/IL21 and IRES/GM-SCF-IL21 groups respectively.The survival rate of mice at 60 days of treatment was 73.33%, 13.33%, and 6.67% for groups IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21, respectively.The survival rate of the mice was significantly higher in IRES/GM-SCF-IL21 than the other groups.The levels of IL-2 and INF-γof mice 1-6 days after treatment gradually increased in the IRES/combination groups, including IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21.They were highest in the IRES/combination group and lowest (P<0.01) in the IRES/GM-SCF and IRES/IL21 groups, with the IRES/GM-SCF-IL21 group showing intermediate levels.By 6-10 days after treatment, IL-2 and INF-γlevels had stably increased in the IRES/combination groups, but had gradually decreased in the IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21 groups.At the end of treatment, IL-2 and INF-γlevels were significantly (P<0.01) higher in the IRES/GM-SCF-IL21 than were found in either the IRES/GM-SCF group or IRES/IL21 group, which were also significantly (P<0.01) higher than either the IRES/GFP or control groups.The levels of IL-2 and INF-γwere highest in the IRES/combination group ( P<0.01) and not significantly different among the IRES/GM-SCF, IRES/IL21, IRES/GFP, and control groups.Conclusion The inhibitory effects in subcutaneous liver cancer model in mice were obvious significantly, and its mechanism maybe be related to the activation of the body's immune.
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Objective Interleukin-21(IL-21) is likely to contribute to the development of liver fibrosis, but up to now, no study has been reported on the relationship between IL-21 and intrauterine adhesions ( IUA) .This study aimed to establish a rat model of IUA induced by mechanical injury and lipopolysaccharide (LPS) infection (dual injury), determine the expression level of serum IL-21, and confirm the association of serum IL-21 with the formation of IUA. Methods Forty healthy female SD rats were randomly divided into four groups of equal number,control, mechanical injury, LPSinfection, and dual injury.At 7 days after IUA modeling,uterine tissue-swere collected from all the animals for observation of the endometrial glands,detection of the degree of IUA by Masson staining, measure-ment of the serum IL-21 level by radioimmunoassay, and analysis of the correlation between the number of endometrial glands and the de-gree of fibrosis. Results The number of endometrial glandswas significantly smallerin the dual injury group (3.59±1.20) than in the mechanical injury (11.66±2.34) and LPSinfection group(11.59±1.47)(P<0.05), while the proportion of fibrosis area wassignificantly higher in the former group(0.65±0.03) than in the lattertwo(0.30±0.07 and 0.32±0.08)(P<0.05).The level of serum IL-21 was signifi-cantly increased in the dual injury group ([286.21±27.80]pg/mL) as compared with those in the control ( [ 118.65 ±22.55 ] pg/mL ) , mechanical injury([176.20±19.05]pg/mL), and LPS infection group ([187.98±16.51]pg/mL) (P<0.05), with a positive correlation be-tween the IL-21 level and theproportion of fibrosis area ( r=0.271, P<0.05) . Conclusion A rat model ofIUAwas successfully established by mechanical injury and lipopolysaccharide (LPS) infection.The evi-dent increase of serum IL-21 in the IUA model was positively correlated with the percentage of fibrosis area, suggesting that IL-21 may be involved indirectly in the formation of IUA.
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Objective:To explore the expression of IL-21 and the relationship with adenine nucleotide translocator antibody, ( ANT antibody ) in different courses of viral myocarditis ( VMC ) mice, and to explore the role and significance of IL-21 in VMC.Methods:To induced VMC mice model, BALB/c male mice were peritoneally injected ( IP ) with Coxsackievirus B3, mice peritoneally injected with PBS were taken as the controls.On 0,1,2,3,4 and 6 w after IP,the level of IL-21 mRNA in mice myocardic tissue was determined by real-time RT-PCR.The expression of IL-21 protein and ANT antibody in serum was evaluated by EILSA.Results:Comparing with the controls,the expression of IL-21 mRNA and protein increased from 1st week after IP,reaching the peak on the 2nd week,and maintaining a higher trend until 6th week.All the results at each time except that of 0 week were higher than those of controls(P<0.05).The levels of ANT antibody were also increased from 1st to 6th week,reaching the peak on the 2nd week all the results at each time except that of 0 week were higher than those of controls(P<0.05).Pearson analysis showed that there was rela-tionship between IL-21 and ANT antibody( r=0.88,P<0.01).Conclusion:The datas show that local significantly increased levels of IL-21 in myocardium and serum, and have positive relationship with the level of adenine nucleotide translocator antibody, which indicates that IL-21 may play a role in the pathogenesis of mice virus myocarditis.
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Objective To test the level of follicular helper T cells (Tfh) and interleukin (IL)-21,CXCL13 in the peripheral blood of patients with ankylosing spondylitis (AS),and to analyze the relationship between Tfh and clinic features and explore the possible immunological pathogenesis of AS.Methods The Tfh cells were obtained from patients and normal controls and detected by flow cytometry.While the levels of IL-21,CXCL13 in patients and normal controls were measured by enzyme-linked immunosorbent assay (ELISA) tests.Data analysis were performed by Student's t-test,Rank-sum test,Spearman's correlation test.Results The expression of CD4+CXCR5 qCOS + cells (Tfh) (mean rank 33.71) and IL-21 [(299±27) ng/L],CXCL13 [(5.8±1.0) μg/L] in the peripheral blood of AS was significantly higher than normal controls [mean rank 23.54,(176±26) ng/L,(4.2±0.8) μg/L] (Z=-2.258,t=17.221,t=6.464,all P＜0.05).It was similar in AS with peripheral joint involvement compared with AS of non-peripheral joint involvement,and there was no difference between AS patients with positive HLA-B27 and those without HLA-B27.Mean -while,no correlation was found between the expression of Tfh,IL-21,CXCL13 and level of ESR,CRP,BASDAI.And there was no significant correlation between the expression of Tfh and IL-21,CXCL13 (P＞0.05).Conclusion The expression of Tfh and the levels of IL-21,CXCL13 are increased significantly,but are not closely relatedto disease activity.These results indicate that the abnormality of Tfh may play an important role in the pathogenesis of AS.
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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.