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.     

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.     

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。     

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.     

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.     

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.     

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.     

Th17细胞介导肿瘤免疫的双向性及其意义

Th17细胞作为众多CD4+辅助性T细胞的一种,其标志性特点是能分泌炎性因子白细胞介素(IL-17)参与炎症反应.随着人们对炎症的深入了解,发现Th17细胞不只是发挥炎性效应,其在肿瘤的发生、发展及抗肿瘤方面也有重要的作用.研究发现,在多种肿瘤中存在Th17细胞及其相关的细胞因子,Th17细胞分泌炎性因子IL-17主动增强相关的抗肿瘤信号通路,调控抗肿瘤免疫应答.另外,固有免疫细胞IL-17+γδT细胞以及IL-17+ Foxp3T细胞不仅分泌IL-17发挥促肿瘤效应,还能协同Th17细胞参与到抗肿瘤免疫反应中,共同调控机体的抗肿瘤免疫反应.因而研究Th17细胞参与肿瘤应答的机制以及与IL-17+γδT细胞和IL-17+ Foxp3T细胞在肿瘤免疫中的相互作用关系具有重要意义.     

Th17和 Treg 细胞及其相关细胞因子在结核性胸膜炎患者中的变化及意义

目的检测健康人和结核性胸膜炎患者外周血Th17细胞和调节性T细胞（ Treg细胞）（ CD4＋CD25＋Foxp3＋）在CD4＋T细胞中的表达率以及IL-17、IL-23、IL-6、TGF-β血清水平和患者胸水中的IL-17、IL-23、IL-6、TGF-β水平,研究Th17细胞和调节性T细胞以及IL-17、IL-23、IL-6、TGF-β在结核性胸膜炎发病机制中的作用。方法使用流式细胞术检测患者以及健康对照人群外周血Th17细胞和调节性T细胞表达率,ELSIA方法定量检测血清以及胸水中IL-17、IL-23、IL-6、TGF-β水平,使用SPSS17．0统计学软件,分析健康人和结核性胸膜炎患者上述指标之间的差异以及各指标间的相关性。结果结核性胸膜炎患者外周血Th17细胞表达率（1．02％±0．20％）明显高于健康人外周血Th17细胞表达率（0．89％±0．13％,P＝0．002＜0．05）;结核性胸膜炎患者外周血调节性T细胞表达率（4．64％±0．77％）明显低于健康人外周血调节性T细胞表达率（5．10％±0．90％,P＝0．000＜0．05）;结核性胸膜炎患者Th17/Treg细胞的比率（0．25±0．07）明显高于健康人（0．17±0．05,P＝0．000＜0．05）;结核性胸膜炎患者外周血IL-17（17．49 ng/L±3．94 ng/L）和IL-23（90．42 ng/L±23．06 ng/L）水平和胸水中IL-17（26．13 ng/L±5．98 ng/L）和IL-23（122．26 ng/L±31．71 ng/L）水平显著高于对照组外周血IL-17（14．45 ng/L±3．81 ng/L）和IL-23（77．55 ng/L±20．26 ng/L）的水平,P值分别为0．022、0．039、0．000、0．000;患者胸水中IL-17和IL-23浓度也显著高于本人血液中的IL-17和IL-23浓度,P值为0．000和0．000;患者胸水中IL-6的浓度（5．31 ng/L±0．74 ng/L）显著高于患者血液中IL-6的浓度（4．54 ng/L±1．02 ng/L）和对照组血液中IL-6的浓度（4．26 ng/L±0．91 ng/L）,P值分别为0．003和0．000,患者血液与对照组血液中IL-6的浓度没有显著差别（P＝0．274）;对照组外周血液TGF-β浓度（3．95 ng/L±0．79 ng/L）显著高于患者外周血液TGF-β浓度（3．32 ng/L±0．80 ng/L）及胸水中TGF-β浓度（3．12±0．77）,P值分别为0．005和0．000,患者血液及胸水中TGF-β水平之间没有显著差别（P＝0．365）;结核性胸膜炎患者外周血Th17细胞的表达率与其Treg细胞在外周血的表达率呈显著负相关（r＝-0．684, P＝0．000＜0．05）,结核性胸膜炎患者外周血Th17细胞的表达率与其外周血中的IL-17、IL-23、IL-6水平呈明显的正相关（r＝0．479,0．441,0．326,P＝0．013,0．015,0．017）;患者血液中TGF-β水平与Treg细胞在外周血的表达率呈明显的正相关（r＝0．297,P＝0．024）,与Th17细胞表达率没有明显相关性（r＝0．091,P＝0．659）。结论 Th17和Treg细胞可能参与了结核性胸膜炎的免疫病理机制,有关细胞因子的变化可能参与了Th17和Treg细胞变化的调控以及炎症反应, Th17和Treg细胞以及有关细胞因子的变化可能是结核性胸膜炎重要的免疫病理机制。     

强直性脊柱炎患者外周血中Th17细胞水平研究

目的建立流式细胞术检测外周血Th17的方法,并探讨强直性脊柱炎患者外周血Th17变化的意义。方法采用四色流式细胞术检测16例AS患者(AS组)和16例健康志愿者(对照组)外周血Th17细胞比例。结果用流式细胞仪结合不同荧光素标记的CD3、CD8、CD45和IL-17A单克隆抗体可准确检测外周血中Th17细胞的数量。对照组中Th17的检测结果分别为(1.58±1.09)%;AS患者组中分别为(3.06±2.24)%,AS患者外周血中Th17细胞显著高于对照组(P=0.004)。结论患者外周血中Th17细胞数量与对照组比较显著升高,该种细胞可能参与了AS的发生和发展。     

ESAT-6- and CFP-10-specific Th1, Th22 and Th17 cells in tuberculous pleurisy may contribute to the local immune response against Mycobacterium tuberculosis infection

Th1 cell-mediated adaptive immune response is very important but may not be sufficient to control Mycobacterium tuberculosis (M. tuberculosis) infection. The roles of the various T cell subsets and cytokines in the inflammatory processes are not clearly elucidated. We investigated whether Th1, Th22 and Th17 cells mediated cellular immunity at the local site of M. tuberculosis infection in patients with tuberculous pleurisy (TBP). The results showed that the cytokines IFN-γ and IL-22 but not IL-17 were elevated in tubercular pleural fluid. Following stimulation with immune-dominant peptides of early secreted antigenic target-6 (ESAT-6), culture filtrate protein-10 (CFP-10) or Bacille Calmette-Guerin, pleural fluid mononuclear cells expressed high levels of cytokines IFN-γ, IL-22 and IL-17 as revealed by mRNA and protein measurements. In addition, we showed that cytokines IFN-γ, IL-22 and IL-17 were produced in M. tuberculosis-specific immune response by distinct subsets of CD4+ T cells with the phenotype of CD45RA-CD62L-CCR7+CD27+ . Our results demonstrated for the first time that ESAT-6- and CFP-10-specific Th1, Th22 and Th17 cells existed in the patients with TBP and might play an essential role against M. tuberculosis infection. The findings of this study raised the possibility of unravelling the critical targets for therapeutic intervention in chronic inflammatory diseases such as TBP.     

Bordetella pertussis infection induces a mucosal IL-17 response and long-lived Th17 and Th1 immune memory cells in nonhuman primates

Despite near universal vaccine coverage, the bacterial pathogen Bordetella pertussis has re-emerged as a major public health concern. We recently developed a baboon (Papio anubis) model of pertussis that provides an excellent model of human pertussis. Using this model, the immune response to pertussis was characterized by measuring cytokines in the nasopharyngeal mucosa of infected baboons. Notably, we observed mucosal expression of interleukin-17 (IL-17) as well as IL-6, IL-23, and several cytokines and chemokines that are orchestrated by IL-17 immune responses. We also found substantial populations of circulating B. pertussis-specific Th17 and Th1 cells in convalescent animals >2 years post-infection consistent with a role in immunological memory to pertussis. Collectively, these data shed important light on the innate and adaptive immune responses to pertussis in a primate infection model and suggest that Th17 and Th1 immune responses contribute to the immunity conferred by natural pertussis infection.