流式细胞术检测慢性乙型肝炎患者外周血Th17和T_C17细胞

目的建立流式细胞术检测外周血中Th17、TC17细胞的方法,并探讨慢性乙型肝炎(CHB)患者外周血Th17细胞、TC17细胞变化的的意义。方法用三色流式细胞术检测了24例CHB患者和15例健康对照者外周血中Th17细胞和TC17细胞。结果用流式细胞仪结合不同荧光素标记的CD3、CD8和IL-17A单克隆抗体可准确检测外周血中Th17、TC17细胞的数量。对照组中Th17、TC17的检测结果分别为(2.13±0.50)%(1.6%～3.3%)和(0.27±0.13)%(0.2%～0.6%);CHB患者组中分别为(5.58±4.52)%(1.6%～19.0%)和(1.05±0.93)%(0.2%～3.1%)。CHB患者外周血中Th17、TC17细胞显著高于对照组(P=0.007、P=0.004)。结论流式细胞术检测外周血Th17、TC17细胞方法可靠;CHB患者外周血中Th17细胞、TC17细胞与对照组比较显著增高。     

桥本甲状腺炎患者外周血滤泡辅助T细胞水平及临床意义

目的 检测桥本甲状腺炎(HT)患者外周血滤泡辅助性T细胞(follicular helper T cells,Tfh)水平,探讨Tfh与HT发病的关系.方法 采用电化学发光法检测HT患者及对照组外周血FT3、FT4、TSH、甲状腺自身抗体水平,采用流式细胞术分析各组试验对象的外周血Tfh的水平,同时进行相关性分析.结果 与对照组相比,桥本甲状腺炎患者外周血Tfh比例显著升高(P＜0.05);桥本甲状腺炎患者外周血Tfh与甲状腺自身抗体TPO-Ab呈正相关(P＜0.05),但与Tg-Ab无相关性(P＞0.05).结果 HT患者外周血中Tfh可能参与桥本甲状腺炎的发生和发展.     

肺结核患者外周血Th17/Treg的表达及其病理意义

为探讨肺结核患者外周血Th17与Treg的表达及其病理意义,收集2017年1月至2018年6月青海省第四人民医院收治的80例肺结核患者作为结核组,同期收集80例健康成人作为对照组。观察两组外周血Th17与Treg及其细胞因子的表达情况,同时分析Th17与Treg及其细胞因子与肺结核患者病情严重程度的相关性。结果显示,与对照组比较,结核组Th17降低[(2.69±0.93)%vs(1.94±0.91)%,P=0.000],Treg升高[(7.88±2.65)%vs(9.60±2.59)%,P=0.000],IL-17降低[(32.89±8.92) pg/mL vs(25.72±9.62) pg/mL,P=0.000],IL-10升高[(22.80±6.81) pg/mL vs(26.61±7.62) pg/mL,P=0.001]。肺结核患者病灶数目、直径总和与Th17、IL-17呈显著负相关(P0.05),与Treg、IL-10呈显著正相关(P0.05)。与无空洞形成的患者比较,空洞形成的患者Th17降低[(2.06±0.62)%vs(1.76±0.68)%,P=0.045],Treg升高[(8.79±2.12)%vs(10.81±2.41)%,P=0.000],IL-17和IL-10差异无统计学意义(P0.05)。提示外周血中Th17与Treg在肺结核患者中表达异常,与结核病情严重度相关。     

慢性肝病患者外周血淋巴细胞亚群的变化

本文采用流式细胞仪对５１例慢性肝病患者外周血淋巴细胞亚群进行了测定。结果发现慢性活动性肝为（ＣＡＨ）、慢性迁延性肝炎（ＣＰＨ）和肝炎后肝硬化（ＨＣ）患者与正常对照相比，ＮＫ数量明显增加（Ｐ〈０．０１），Ｄ４＾＋和ＣＤ３＾＋数量均明显降低（Ｐ〈０．０１），ＣＡＨ的ＣＤ８＾＋数量明显增加（Ｐ〈０．０１），且ＣＤ４＾＋／ＣＤ８＾＋比值明显下降（Ｐ〈０．０１），ＨＣ的ＣＤ４＾＋／ＣＤ８＾＋比值下降（Ｐ〈０     

Treg/Th17与慢性阻塞性肺疾病的研究进展

慢性阻塞性肺疾病(chronicobstructivepulmonarydisease,COPD)是气道的炎性病症,以慢性进展和不可逆的气流受限为特征.研究表明自身免疫反应参与本病的发病机制.调节性T细胞(regulatoryT-cells,Treg)和辅助性T细胞17(Thelper17cells,Th17)是CD4+T淋巴细胞的两个亚群,是已知的重的免疫调节细胞,牵涉到多种炎性和自身免疫性疾病的发病机理,并在气道炎症和慢性阻塞性肺病的发病机制中起到重作用.两在功能和分化过程中相互拮抗,该平衡关系的维持或破坏对于免疫或炎症性疾病的发病具有重意义.     

哮喘-慢性阻塞性肺疾病患者外周血Treg/Th17失衡的研究

为探究哮喘-慢性阻塞性肺疾病(asthma-chronic obstructive pulmonary disease overlap,ACO)患者外周血 Treg/Th17失衡对疾病的诊断价值,收集于阜阳市第二人民医院就诊的ACO患者52例(ACO组),哮喘急性发作期患者34例(哮喘组),慢性阻塞性肺疾病(chron...     

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。     

γδT细胞、Th17细胞和IL-17在慢性乙型肝炎患者外周血中表达的意义

目的:了解γδT细胞、Th17细胞和IL-17在慢性乙型肝炎（CHB）患者外周血中的表达情况,探讨γδT细胞、Th17细胞和IL-17在CHB发病机制中可能的作用。方法:随机选取2015年11月至2016年4月本院门诊或住院收治的慢性无症状HBV携带者（AsC）20例、CHB轻度患者20例、CHB中和重度患者20例、同...     

子宫内膜异位症患者腹腔Th17细胞和Th17.1细胞检测及致炎活性分析

目的:鉴别和分离子宫内膜异位症患者腹腔冲洗液中的辅助性T细胞17(Th17)和辅助性T细胞17.1(Th17.1),并比较二者致炎活性。方法:收集中重型子宫内膜异位症患者的腹腔冲洗液,根据细胞表面趋化因子受体CC趋化因子受体6(CCR6)、CC趋化因子受体10 (CCR10)、CC趋化因子受体4(CCR4)、CXC趋化因子受体3(CXCR3)表达差异,利用流式细胞术鉴别不同的腹腔T细胞亚群,检测其白介素-17A(IL-17A)、干扰素-γ(IFN-γ)和T-bet转录因子(T-bet)的表达,同时结合实时定量逆转录聚合酶链反应检测RAR相关孤儿受体C (RORC)、肿瘤坏死因子-α(TNF-α)、粒细胞-巨噬细胞集落刺激因子(CSF2)、白介素1受体Ⅰ型(IL1R1)等基因的mRNA水平,鉴别Th17和Th17.1细胞,并比较其致炎活性的差异。结果:腹腔Th17细胞的表型为CD3⁺CD4⁺CCR6⁺CCR10^-CCR4⁺CXCR3^-/lo, Th17.1细...     

类风湿性关节炎患者外周血Th17细胞/调节性T细胞比例增加并与疾病活动相关

目的通过检测类风湿性关节炎(RA)患者外周血Th17细胞、调节性T细胞(Treg)的相对数量,探讨其在RA发病机制中的作用及其临床意义。方法选取符合入组标准的RA患者60例及健康体检者15例,采用流式细胞术检测Th17细胞/Treg占外周血CD4+细胞百分比,比较其在RA患者与健康体检者间的差异,分析其表达与肿胀关节数、压痛关节数、28个关节疾病活动度评分(DAS28)、疼痛视觉模拟评分(VAS)、红细胞沉降率(ESR)、C反应蛋白(CRP)的关系。结果与健康对照组比较,RA组患者外周血Th17细胞数量明显增加、Treg数量明显减少。RA患者外周血中Th17细胞数量与肿胀关节数、压痛关节数、DAS28分值及CRP呈正相关;RA患者外周血中Treg的数量与肿胀关节数、压痛关节数、疼痛VAS、DAS28分值、ESR及CRP等病情活动指标无明显相关性。结论 RA患者外周血Th17细胞增加,Treg减少,以Th17占优势,Th17细胞数量增加与RA病情活动有关。     

Age-associated alteration in naive and memory Th17 cell response in humans

Th17 cells produce IL-17 that plays an important role in host defense. However, little is known about whether aging affects human Th17 cells. Here we demonstrated that healthy elderly people (age ≥ 65) had a decreased frequency of IL-17-producing cells in memory CD4(+) T cells compared to healthy young people (age ≤ 40) while both groups had similar frequencies of IFN-γ-producing cells in the same memory cell subset as measured by flow cytometry. In contrast, the healthy elderly had increased differentiation of IL-17-producing effector cells but not IFN-γ-producing cells from naive CD4(+) T cells compared to the healthy young. The results of ELISA also showed similar findings with increased IL-17 production from naive CD4(+) T cells and decreased IL-17 production from memory CD4(+) T cells in the elderly compared to the young. These findings indicate that aging differentially affects naive and memory Th17 cell responses in humans.     

Complement activation by malignant B cells from patients with chronic lymphocytic leukaemia (CLL).

It has previously been reported that the expression of the complement receptors CR1 (CD35) and CR2 (CD21) on malignant B cells in CLL is reduced compared with the expression on normal B cells, while deposition of complement C3 fragments, as a consequence of alternative pathway (AP) activation of complement, is observed on mononuclear cells from patients with B CLL. Following our demonstration that normal B cells are capable of activating the AP of complement in a CR2-dependent fashion, we have chosen to re-examine the complement-activating ability of B CLL cells in relation to their altered phenotype with respect to CR2 and the complement regulatory membrane proteins, CR1, decay accelerating factor (DAF) (CD55) and membrane cofactor protein (MCP) (CD46). Flow cytometry was used to measure expression of complement receptors and regulatory proteins on CD5+ B cells from CLL patients, as well as the deposition of C3 fragments occurring both in vivo and after in vitro AP activation. We have confirmed the reduced expression of CR1 and CR2 on CLL cells and have shown that AP activation in the presence of homologous, normal serum was reduced on B CLL cells compared with normal B cells. The degree of AP activation correlated directly with CR2 expression. In addition, we observed that CLL cells bear in vivo-deposited C3d,g, although at a significantly lower level than normal B cells.     

Blood dendritic cells in patients with chronic lymphocytic leukaemia

Myeloid and plasmacytoid dendritic cells (MDC, PDC) play a key role in the initiation of immune responses. We found a reduction of both DC subsets in 42 patients with chronic lymphocytic leukaemia (CLL) at diagnosis (P<0.0001 and 0.0001 vs. controls, respectively), likely related to the high secretion of CCL22 and CXCL12 (P=0.04 and 0.008 vs. controls, respectively) by leukaemic cells. However, CD14+ monocytes from CLL patients could give rise to functional IL-12p70-secreting monocyte-derived DCs, capable of inducing a type 1 polarization immunostimulatory profile. These monocyte-derived DCs from CLL patients efficiently migrate in response to CCL19/MIP-3beta chemokine, suggesting that functional autologous DCs can be generated for immunotherapeutic purposes to circumvent DC defects in CLL.     

过敏性哮喘患者调节性T细胞对Th17细胞和Th9细胞的影响

目的:深入研究过敏性哮喘病人(轻到中度)急性发作期的调节性T细胞对Th17和Th9细胞功能影响的异常。方法:招募30例过敏性哮喘病人(轻到中度)急性发作期和30例健康者,采集外周静脉血30 ml,分离PBMC,然后应用磁珠法体外分离CD4+CD25+CD127-/low调节性T细胞(Tregs)和CD4+CD25-效应T细胞(选取分离纯度90%以上的标本继续下一步试验)。在PHA刺激下体外培养,检测效应T细胞增殖情况和Th17细胞、Th9细胞特异性基因(RORC和PU.1)表达及特异性细胞因子(IL-17和IL-9)分泌情况,检测哮喘病人Th17细胞和Th9细胞的异常情况;并给予Tregs干预,检测Tregs对Th17细胞和Th9细胞分化的影响。结果:哮喘组和健康对照组的Tregs均可以抑制CD4+CD25-效应细胞增殖反应,但哮喘组Tregs的抑制功能较健康对照组明显下降(P=0.03)。哮喘病人RORC表达和IL-17分泌均高于对照组(P0.05),Tregs对RORC表达的抑制能力在哮喘组有所下降(P0.05),但对IL-17分泌的抑制能力在哮喘组和对照组没有显著差别(P0.05);哮喘病人IL-9分泌无论是否加入Tregs干预均高于对照组(P0.05),但PU.1表达只有在加入Tregs干预后才高于对照组(P=0.04);Tregs对PU.1表达和IL-9分泌的抑制能力在哮喘组和对照组没有显著差别(P0.05)。结论:过敏性哮喘病人(轻到中度)急性发作期Tregs数量和功能下降,而Th17细胞和Th9细胞的特异性基因表达和特异性细胞因子生成增加,哮喘病人的Tregs在体外对Th17细胞的抑制功能有所下降,但尚未发现Tregs对Th9细胞的抑制功能明显降低。     

川崎病急性期患者Th17细胞检测及意义

目的:探讨川崎病患者急性期Th17细胞的水平及意义。方法:急性期川崎病(KD)患儿43例,正常同年龄对照41例,采用流式细胞术检测外周血单个核细胞(PBMC)中Th17细胞占CD4+细胞的比例。荧光定量PCR(Real-time-PCR)检测RORγt和IL-23p19mRNA的表达。双抗体夹心酶联免疫吸附法(ELISA)检测血清IL-6水平。结果:急性期KD患儿外周血Th17细胞比例及其特异性转录因子RORγt的mRNA表达分别为:(1.285±0.625)%、(M:5.62,Q:7.16),显著高于同龄对照组(0.584±0.407)%、(M:1.79,Q:2.55),(P0.01)。KD患儿血清IL-6水平显著增高[病人:(M:36.42,Q:129.77)、对照:(M:2.39,Q:1.15)](P0.01),且与Th17细胞比例成正相关(r=0.88,P0.01)。IL-23p19的mRNA表达与正常对照比较无显著差异[病人:(M:2.12,Q:3.05)、对照:(M:1.73,Q:2.79)](P0.05)。结论:急性期川崎病患者Th17细胞数量、血清IL-6水平升高;Th17可能作为急性炎症的启动因素,参与川崎病免疫发病过程。     

T cell‐derived leptin contributes to increased frequency of T helper type 17 cells in female patients with Hashimoto's thyroiditis

Leptin modulates T cell function and plays an important role in autoimmune diseases. Our study aimed to explore the role of leptin and T helper type 17 (Th17) cells in Hashimoto's thyroiditis patients. Twenty‐seven patients with Hashimoto's thyroiditis (HT) and 20 healthy controls were enrolled into the current study. A modest increase of plasma leptin in HT patients and the CD4⁺ T cell‐derived leptin from HT patients was stronger than that from healthy controls. In HT patients, there are no statistically significant correlations between plasma leptin concentrations and the percentage of Th17 cells or the level of retinoic acid‐related orphan receptor γt (RORγt), but strong positive correlations were observed between CD4⁺ T cell‐derived leptin and the percentage of Th17 cells or the level of RORγt mRNA, and additionally significantly up‐regulated leptin, interleukin (IL)17 and RORγt mRNA levels in the thyroid tissue. Furthermore, neutralization of leptin decreases the frequency of Th17 cells in vitro. Current study has revealed an increased leptin involvment in Hashimoto's thyroiditis associated with an increased number of Th17 cells.     

Altered balance between self‐reactive T helper (Th)17 cells and Th10 cells and between full‐length forkhead box protein 3 (FoxP3) and FoxP3 splice variants in Hashimoto's thyroiditis

T helper type 17 (Th17) cells play a pathogenic role in autoimmune disease, while interleukin (IL)‐10‐producing Th10 cells serve a protective role. The balance between the two subsets is regulated by the local cytokine milieu and by the relative expression of intact forkhead box protein 3 (FoxP3) compared to FoxP3Δ2, missing exon 2. Th17 and Th10 cell differentiation has usually been studied using polyclonal stimuli, and little is known about the ability of physiologically relevant self‐antigens to induce Th17 or Th10 cell differentiation in autoimmune thyroid disease. We subjected mononuclear cells from healthy donors and patients with Hashimoto's thyroiditis (HT) or Graves' disease (GD) to polyclonal stimulation, or stimulation with human thyroglobulin (TG), human thyroid peroxidase (TPO), or Esherichia coli lipopolysaccharide (LPS). TPO and LPS induced increased differentiation of naive CD4⁺CD45RA⁺CD45R0^– T cells from HT patients into Th17 cells. Th10 cell proportions were decreased in HT after polyclonal stimulation, but were comparable to those of healthy donors after antigen‐specific stimulation. Taken together, our data show that an increased Th17 : Th10 ratio was found in HT patients after stimulation with thyroid‐specific self‐antigens. We also observed an elevated baseline production of IL‐6 and transforming growth factor (TGF)‐β1 and of mRNA encoding FoxP3Δ2 rather than intact FoxP3. This may contribute to the skewing towards Th17 cell responses in HT.