原发免疫性血小板减少症患者外周血Th22细胞水平的变化及意义

目的:观察Th22细胞在原发免疫性血小板减少症(ITP)患者外周血中的变化.方法:用流式细胞术分别检测ITP初诊组、治疗后完全反应(CR)组及正常对照组外周血中Th22细胞所占比例.用酶联免疫吸附实验(ELISA)分别检测各组外周血中IL-22、TGF-β、TNF-α、IL-6的表达水平.用半定量RT-PCR分别检测各组外周血中IL-22 mRNA的表达水平.并逐层分析比较.结果:ITP初诊组和治疗后组Th22细胞比例、外周血中IL-22、TNF-α、IL-6的表达水平以及IL-22mRNA的表达水平均明显高于正常对照组(P＜0.01),治疗后组Th22细胞比例、IL-22、TNF-α、IL-6、IL-22 mRNA的表达水平均低于初诊组(P＜0.05);ITP初诊组和治疗后组TGF-β水平分别为(3.27±1.02)ng/L、(5.41±1.69)ng/L明显低于正常对照组(9.65±2.78) ng/L(P＜0.01),且初诊组TGF-β的水平低于治疗后组(P＜0.05).结论:ITP患者体内Th22细胞数量增加、TNF-α、IL-6的表达升高、TGF-β的表达减低.     

Elevated Th22 Cells Correlated with Th17 Cells in Patients with Rheumatoid Arthritis

Background

T-helper (Th) 22 and Th17 cells are implicated in the pathogenesis of autoimmune diseases. The roles of Th22 cells in the pathophysiology of rheumatoid arthritis (RA) remain unsettled.

Materials and Methods

CD4⁺IFN??^?IL17^?IL-22⁺ T cells (Th22 cells), CD4⁺IFN??^?IL-22^?IL17⁺ T cells (pure Th17 cells), CD4⁺IL17⁺ T cells (Th17 cells), and CD4⁺IFN??⁺ T cells (Th1 cells) in RA, osteoarthritis patients, and healthy controls were examined by flow cytometry. Plasma IL-22 and IL-17 levels were examined by enzyme-linked immunosorbent assay.

Results

Th22 cells, pure Th17 cells, Th17 cells, and interleukin-22 were significantly elevated in RA patients compared with osteoarthritis and healthy controls, but there were no significant differences regarding Th1 cells and interleukin-17. Th22 cells showed a positive correlation with interleukin-22 as well as pure Th17 cells or Th17 cells in RA patients. Additionally, the percentages of Th22 cells, pure Th17 cells as well as Th17 cells correlated positively with both C-reactive protein levels and 28-joints disease activity score.

Conclusion

Together, our results indicated a possible role of Th22 pure Th17 cells and Th17 cells in RA, and blockade of the interleukin-22 may be a reasonable therapeutic strategy for RA.     

Elevated levels of Th17 cells and Th17-related cytokines are associated with disease activity in patients with inflammatory bowel disease

Objective

Interleukin-17(IL-17)-producing T helper(Th)17 cells are considered as a new subset of cells critical to the development of inflammatory bowel disease (IBD). We aimed to investigate the distribution of Th17 cells, the expressions of Th17-related cytokines (IL-17, IL-21 and IL-22) and their association with disease activity in IBD patients.

Methods

We collected intestinal tissue biopsies from 40 patients with active ulcerative colitis (UC), 20 patients with active Crohn’s disease (CD) and 20 healthy controls. The distribution of Th17 cells and expressions of Th17-related cytokines in colonic tissues were evaluated by a standard immunohistochemical procedure. Serum IL-17, IL-21 and IL-22 levels were determined by ELISA. Pearson’s and Spearman’s correlation analyses were performed to analyze the correlation between the number of Th17 cells, the expressions of Th17-related cytokines and disease activity index, endoscopic and histological grading, and CRP and PLT levels, respectively.

Results

Compared with healthy controls, the number of Th17 cells and the expressions of IL-17, IL-21 and IL-22 were significantly increased in active IBD patients (P < 0.05). In addition, Pearson’s and Spearman’s correlation analyses showed that the number of Th17 cells and the expressions of Th17-related cytokines were correlated with disease activity index, endoscopic and histological grading, CRP and PLT levels (P < 0.05).

Conclusions

Th17 cells and Th17-related cytokines (IL-17, IL-21 and IL-22) were increased in the intestinal mucosa in active IBD patients and may play an important role in disease activity and mucosal damage.     

Increased number of Th22 cells and correlation with Th17 cells in peripheral blood of patients with IgA nephropathy

T-helper (Th) 22 and Th17 cells are involved in the pathogenesis of autoimmune diseases. However, the role of Th22 and correlation with Th17 cells in the pathophysiology of IgA nephropathy (IgAN) remain unknown. In our study, Th22 and Th17 cells in peripheral blood of IgAN patients, non-IgA mesangial proliferative glomerulonephritis (non-IgA MsPGN) patients, and healthy controls were measured by flow cytometry. The concentration of plasma interleukin-22 (IL-22) was examined by enzyme linked immunosorbent assay (ELISA). The results showed that Th22 cells, Th17 cells, and plasma IL-22 were significantly elevated in IgAN patients compared with non-IgA MsPGN patients and healthy controls. Th22 cells showed a positive correlation with the levels of plasma IL-22 in IgAN patients. Moreover, a significantly positive correlation between Th22 cells and Th17 in IgAN patients was observed. Furthermore, IgAN patients with proteinuria showed a higher percentage of Th22 cells than IgAN patients without proteinuria. Our data demonstrated that IgAN had increased frequencies of peripheral Th22, Th17 cells and plasma IL-22, indicating that Th22 along with Th17 cells are involved in the immune responses of IgAN.     

CCR6 defines a subset of activated memory T cells of Th17 potential in immune thrombocytopenia

Current researches have determined the significance of C-C chemokine receptor (CCR)6 expression as either a marker of T helper cells (Th) or an effector and regulator of T cell function. However, the roles of CCR6 in the pathogenesis of immune thrombocytopenia (ITP) are unclear. In this study, we aimed to investigate the phenotype and functional characteristics of circulating CCR6⁺ T cells in blood from chronic ITP patients and healthy controls. We found that the frequency of CCR6⁺CD4⁺ cells was higher in ITP patients than in healthy controls. Anti-CD3/anti-CD28 stimulation induced rapid expansion of CCR6⁺CD4⁺ cells in ITP patients. CCR6⁺CD4⁺ cells had a phenotype of activated cells and predominantly expressed CD45RO. Forkhead box protein P3 (FoxP3) and CD25-positive cells were exclusively detected within the CCR6⁺CD4⁺ cells. In ITP patients, CCR6⁺ regulatory T cells (T_reg) were decreased and positively correlated with platelet counts and transforming growth factor (TGF)-β plasma levels. In contrast to CCR6^– counterparts, CCR6⁺CD4⁺ cells produced higher levels of interleukin (IL)-17A. The frequency of CCR6⁺ Th17 was higher in ITP patients and positively correlated with IL-17A levels in supernatant. Most importantly, CCR6⁺CD4⁺ cell subpopulations, but not CCR6⁻CD4⁺, were closely correlated to treatment response of ITP patients. These findings suggest that circulating CCR6⁺CD4⁺ cells in ITP patients have characteristics of activated memory Th17 phenotype and could be used to monitor disease activity and treatment response.     

COPD 患者Th17 细胞和Treg 细胞介导的免疫应答变化及免疫失衡与肺功能的关系研究

目的:探讨慢性阻塞性肺疾病(COPD)患者外周血Th17 细胞和调节性T 细胞(Treg)及其介导的免疫应答变化与患者肺功能的关系。方法:选取本院呼吸内科收治的90 例COPD 患者(COPD 组)和同期年龄、性别基本匹配的45 例健康体检对象作为对照组,并根据COPD 病情进行亚组分析,分别测定各组研究对象肺功能指标、Th17 细胞、Treg 细胞及血清炎症因子水平。结果:在COPD 患者间,随着病情加重,FEV1%、FVC%、FEV1/ FVC 比值中度COPD 组<重度COPD 组<极重度组(P<0.05);在COPD 患者间,随着病情加重,外周血中Th17 细胞、Th17/ Treg 比值中度COPD 组<重度COPD 组<极重度组(P<0.05),外周血Treg 细胞比值中度COPD 组>重度COPD 组>极重度组(P<0.05),且外周血Th17 细胞、Treg 细胞相关细胞因子变化具有一致性;COPD 患者的肺功能指标FEV1%、FVC%、FEV1/ FVC 与外周血中Th17 细胞、Th17/ Treg 比值呈显著的负相关关系(P<0.05),肺功能指标FEV1%、FVC%、FEV1/ FVC 与外周血Treg 比值呈显著的正相关关系(P<0.05)。结论:COPD外周血Th17/ Treg 比值与患者肺功能下降具有显著的相关性。     

Elevated circulating Th17 and follicular helper CD4+ T cells in patients with rheumatoid arthritis

It remains not fully elucidated the potential functions of Th17 cells and follicular helper T (Tfh) cells and secreting cytokines in the pathogenesis of rheumatoid arthritis (RA) and their association with disease activity. In this study, the frequencies of Th17 and Tfh cells were determined by flow cytometry, and the levels of interleukin (IL)‐17, IL‐21, and IL‐22 were measured by ELISA in RA patients with different disease activities. The dynamic changes of cell subsets were also detected in response to disease‐modify antirheumatic drugs (DMARDs) therapy. The percentages of CD3⁺CD4⁺IL‐17A⁺ (Th17) cells and CD3⁺CD4⁺CXCR5⁺ICOS^high (Tfh) cells, as well as the concentrations of IL‐17, IL‐21, and IL‐22 were significantly elevated in RA patients than those in healthy individuals. Furthermore, Tfh cells, IL‐21, and IL‐22 in the serum was positively correlated with the values of disease activity score. Concentrations of IL‐21 and IL‐22 in the serum were remarkably reduced following the DMARDs therapies. Our data suggested that Th17 cells, Tfh cells as well as the secreting cytokines may be involved in the pathogenesis of RA. The frequency of circulating Tfh cells and the productions of IL‐21 and IL‐22 were associated with the disease activity of RA patients, and might be potential therapeutic targets for treatment of RA.     

支气管哮喘患者外周血Th17 与Th9 细胞的变化及临床意义

目的:探讨Th17、Th9 细胞水平在支气管哮喘患者中的表达及其临床意义。方法:选取67 例哮喘患者为研究对象,30 例健康体检者为对照组,根据全球哮喘防治倡议(GINA)将哮喘患者分为间歇与轻度持续组20 例,中度持续组25 例和重度持续组22 例。收集患者临床资料,完成哮喘控制测试(ACT)评分、肺功能和呼出气一氧化氮(FeNO)值测定,流式细胞仪检测外周血PBMC 中CD3+ CD8- IL-17+ 、CD3+ CD8- IL-9+细胞水平。结果:与健康对照组相比,哮喘患者Th17、Th9 细胞水平均明显升高(P<0.05);中重度哮喘患者Th17 及Th9 细胞水平显著高于间歇与轻度哮喘组(P<0.05),而间歇与轻度哮喘组患者与健康对照组比较,差异无统计学意义(P>0.05)。哮喘患者外周血Th17 及Th9 细胞的表达均与FeNO 值正相关(r 值依次为0.501、0.438;P<0.05),与ACT 评分、肺功能FEV1、FEV1/ FVC 及PEF 负相关(r 值依次为-0.441、-0.362;-0.307、-0.377;-0.419、-0.411;-0.428、-0.24;P<0.05),同时Th17 与Th9 细胞在哮喘患者外周血中的表达率正相关(r 值为0.443,P<0.05)。结论:Th17、Th9 细胞在哮喘患者中高表达,具有协同致炎作用,且与症状严重程度相关,提示Th17 及Th9 细胞对哮喘患者病情评估有潜在的参考价值。     

乙肝相关慢加急性肝衰竭患者Th17细胞水平及其临床意义

目的探讨乙肝相关慢加急性肝衰竭（HBV-ACLF）外周血Th17细胞的表达及其与临床转归的关系。方法采用流式细胞术分析外周血中Th17细胞比例,flowcytomix技术检测血清中IL-17的水平,荧光PCR检测HBV-DNA载量,ELISA检测HBeAg的状态,分析Th17细胞及IL-17的表达与HBV病毒及预后的关系。结果与健康对照组及慢乙肝组相比,HBV-ACLF患者Th17细胞及IL-17水平明显升高（P〈0.01）,且Th17细胞与IL-17水平成正相关;进一步分析发现不同病毒复制水平及HBeAg状态的HBV-ACLF患者Th17细胞及IL-17的差异无统计学意义;与预后好转患者相比,预后不佳HBV-ACLF患者Th17细胞及IL-17水平明显增高（P〈0.05）,且IL-17的水平与终末期肝病评分成正相关。结论 Th17细胞可能参与了HBV-ACLF的发病机制,且IL-17的表达越高可能提示患者的预后不佳。     

Increased Th17 cells and interleukin-17 contribute to immune activation and disease aggravation in patients with chronic hepatitis B virus infection

T helper17 (Th17) cells have been demonstrated to participate in the pathogenesis of hepatitis B virus (HBV) associated liver damage. However, the contribution of Th17 cells to immune activation and disease aggravation in patients with HBV infection is not fully clear. In this study, we investigated the Th17 cells frequencies and interleukin-17 (IL-17) mRNA expressions in peripheral blood mononuclear cells (PBMCs), intrahepatic IL-17-positive cells accumulation, as well as serum IL-17 levels in asymptomatic chronic HBV carriers (AsC), and patients with chronic hepatitis B (CHB) and HBV related acute-on-chronic liver failure (ACLF). Furthermore, the dynamic changes of Th17 cells frequencies and IL-17 concentration in different prognostic ACLF patients were observed. As result, the intrahepatic and peripheral Th17 cells and serum IL-17 concentration were both significantly higher in CHB and HBV related ACLF patients than that in AsC and normal control groups, and increased gradually with immune inflammation aggravation from AsC, CHB to ACLF. Moreover, in ACLF patients, peripheral Th17 cells frequencies were positively correlated with international normalized ratio (INR) and model of end-stage liver disease (MELD) score. Especially the survival patients had an initially lower Th17 cells frequencies and IL-17 levels which gradually decreased following condition improvement as compared with higher baseline level followed by gradually increasing trend in the non-survivals. In conclusion, Th17 cells can be contributed to the immune activation and disease aggravation in patients with chronic HBV infection. This may places Th17 cells as a potential blocking target for controlling CHB and ACLF.     

Elevated interleukin-27 enhances the polarization of Th1/Tc1 cells and the production of proinflammatory cytokines in primary immune thrombocytopenia

Primary immune thrombocytopenia (ITP) is an acquired, organ-specific, autoimmune disease with many immune dysfunctions. Interleukin-27 (IL-27) can regulate T cell differentiation. However, it is unclear whether IL-27 correlates with the dysfunctions of T cell differentiation in ITP patients. Thus, to determine the roles of IL-27 in ITP, we studied the expression of IL-27/IL-27 receptor in ITP patients. The results indicated that the levels of IL-27 in the plasma of untreated active ITP patients were higher than in normal controls. We next evaluated the contribution of IL-27 to T cell differentiation. Our results indicated that IL-27 increased T-bet expression, inhibited GATA-3 and ROR-γt expression, and promoted the secretion of tumor necrosis factor-α, interferon-γ, and granzyme B of peripheral blood mononuclear cells from ITP patients. Also, we confirmed that IL-27 induced the differentiation of T helper (Th)-1 and Tc1 cells. In conclusion, IL-27 might play an important role in the pathogenesis of ITP by inducing the polarization of Th1/Tc1 cells and the production of proinflammatory cytokines.     

Th17细胞在子痫前期患者外周血表达增加

观察正常妊娠妇女和子痫前期疾病患者外周血CD4+T细胞中Th17细胞和特异性转录因子维A酸相关孤独受体(retinoid-related orphan nuclear receptor,RORγt)的表达和意义。实验组为子痫前期疾病患者25例,正常妊娠妇女20例,未孕妇女20例。采集研究对象外周血,酶联免疫吸附试验(ELISA)检测Th17细胞相关细胞因子IL-17A,IL-6,TNF-α的表达,Ficoll法分离外周血单个核细胞(PBMC),免疫磁珠分选CD4+T淋巴细胞,逆转录-聚合酶联反应(RT-RCR)半定量检测CD4+T细胞中Th17细胞特异性转录因子RORγt的表达,流式细胞术检测CD4+IL-17+T细胞比例。子痫前期患者外周血清中IL-6、IL-17A的含量分别为(31.72±13.34)ng/L、(2.61±1.64)ng/L,高于正常妊娠组水平,差异有显著统计学意义(P<0.01),TNF-α在子二组间的表达分别为(18.00±8.64)ng/L和(11.69±3.68)ng/L,差异有统计学意义(P<0.05);RORγt mRNA在子痫前期组的表达高于正常妊娠组,净光密度值差异有显著统计学差异(P<0.01),CD4+IL-17+T细胞在子痫前期组的表达为(1.83±0.42)%,高于正常妊娠组(0.87±0.26)%,差异有显著统计学意义(P<0.01)。子痫前期患者外周血CD4+T细胞中RORγt mRNA、Th17细胞以及Th17细胞相关细胞因子表达异常,可能在疾病的发病机理中起到重要作用。     

骨髓增生异常综合征患者外周血Th22细胞水平的变化及意义

目的:研究Th22细胞在骨髓增生异常综合征(MDS)患者外周血中的表达水平,并探讨其意义.方法:①用流式细胞术分别检测MDS较高危组、较低危组及正常对照组外周血中Th22细胞所占比例.②用酶联免疫吸附实验(ELISA)分别检测各组外周血中IL-22、TGF-β、TNF-α、IL-6的表达水平.③用半定量RT-PCR分别检测各组外周血中Th22细胞的相关转录因子IL-22 mRNA的表达水平,并逐层分析比较.结果:MDS较高危组、较低危组Th22细胞比例、IL-22、TNF-α、IL-6、TGF-β、IL-22 mRNA的表达水平分别为(0.53±0.13)％、(13.51±3.87) ng/L、(146.74±29.74) ng/L、(77.58±16.23) ng/L、(5.21±1.65)ng/L、0.29±0.06和(0.85±0.21)％、(16.13±4.82) ng/L、(182.34±37.55) ng/L、(95.62±20.28) ng/L、(7.46±2.17) ng/L、0.38 ±0.07,均低于正常对照组的(1.24±0.31)％、(19.72±6.55) ng/L、(238.32±50.41) ng/L、(137.95 ±.27.08) ng/L、(9.65±2.78) ng/L、0.49±0.09,差异有统计学意义(P＜0.05),MDS较低危组Th22细胞比例、IL-22、TNF-d、IL-6、TGF-β、IL-22 mRNA的表达水平均高于较高危组,差异有统计学意义(P＜0.05).结论:MDS患者体内Th22细胞数量减少,可能与MDS的发生发展呈负相关,提示MDS患者体内TNF-α、IL-6的表达减低及负向免疫细胞增多可能抑制Th22细胞的分化发育.     

ITP患者脾切除前后外周血Th亚群细胞因子的变化及意义

目的: 探讨辅助性T淋巴细胞(T helper lymphocyte,Th)亚群细胞因子在原发性免疫性血小板减少症(ITP)患者脾切除术前后的表达及意义。方法: 采用QuantiGene Plex (QGP)方法检测22例ITP患者腹腔镜脾切除术前后外周血Th1(IL-2和IFN-γ)、Th2(IL-4、IL-5、IL-6和IL-10)、Th3(TGF-β₁)和Th17(IL-17)细胞因子mRNA表达水平的变化。30例健康体检者作为对照组。结果: (1)术前组IL-2表达水平较对照组明显降低(P<0.05),IL-17表达水平较对照组明显升高(P<0.05),其它细胞因子表达水平在术前组和对照组之间无显著差异(P>0.05)。术后组TGF-β₁表达较术前组和正常对照组均明显升高(P<0.05);术后组IL-2 表达较术前有所升高(P<0.05),但仍低于对照组(P<0.05)。其它细胞因子表达水平在术前组和术后组之间无显著性差异(P>0.05)。(2)术前IL-2与IFN-γ之间成正相关(r=0.647,P<0.01),术前其它细胞因子之间均无明显相关性(P>0.05)。术后3对细胞因子之间成正相关,分别是IL-2与IFN-γ(r=0.787, P<0.01),IL-17与IL-2(r=0.554,P<0.01),IL-17与IFN-γ(r=0.461,P<0.05);术后其它细胞因子之间均无明显相关性(P>0.05)。 结论: ITP患者存在Th亚群细胞因子失衡,脾切除术可改善ITP患者部分Th亚群细胞因子的失衡。     

Elevated Interleukin-21 Correlated to Th17 and Th1 Cells in Patients with Immune Thrombocytopenia

Background  

Interleukin-21 (IL-21) is critical in the development of autoimmune diseases. The role of IL-21 in the pathogenesis of immune thrombocytopenia (ITP) remains unknown.