Frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells correlate with disease activity in rheumatoid arthritis

Abstract

Objective. Rheumatoid arthritis (RA) is a common autoimmune disease that is primarily driven by effector T cells, particularly Th17 cells, which are mainly contained within CD4+CD161+ T cells. Thus, we aimed to explore whether the frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells were correlated with RA disease activity.

Methods. The surface phenotype and cytokine production of blood were analyzed by flow cytometry in 52 RA patients and 17 healthy controls. The disease activity was evaluated by the 28-joint disease activity score.

Results. The frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells were increased in RA patients, and they were elevated in patients with active disease status compared to patients with low disease status. Furthermore, their frequencies were positively correlated with disease activity parameters. Receiver operating characteristic curve analysis revealed that IL-17-producing CD4+CD161+ T cell levels were able to distinguish disease activity with 60.7 % sensitivity and 87.5 % specificity, while CD4+CD161+ T cell levels showed 92.9 % sensitivity and 66.7 % specificity.

Conclusion. These results support the hypothesis that Th17 cells are involved in the pathogenesis of RA and suggest that circulating CD4+CD161+ T cells are a potential biomarker of RA disease activity.     

In silico identified CCR4 antagonists target regulatory T cells and exert adjuvant activity in vaccination

Adjuvants are substances that enhance immune responses and thus improve the efficacy of vaccination. Few adjuvants are available for use in humans, and the one that is most commonly used (alum) often induces suboptimal immunity for protection against many pathogens. There is thus an obvious need to develop new and improved adjuvants. We have therefore taken an approach to adjuvant discovery that uses in silico modeling and structure-based drug-design. As proof-of-principle we chose to target the interaction of the chemokines CCL22 and CCL17 with their receptor CCR4. CCR4 was posited as an adjuvant target based on its expression on CD4(+)CD25(+) regulatory T cells (Tregs), which negatively regulate immune responses induced by dendritic cells (DC), whereas CCL17 and CCL22 are chemotactic agents produced by DC, which are crucial in promoting contact between DC and CCR4(+) T cells. Molecules identified by virtual screening and molecular docking as CCR4 antagonists were able to block CCL22- and CCL17-mediated recruitment of human Tregs and Th2 cells. Furthermore, CCR4 antagonists enhanced DC-mediated human CD4(+) T cell proliferation in an in vitro immune response model and amplified cellular and humoral immune responses in vivo in experimental models when injected in combination with either Modified Vaccinia Ankara expressing Ag85A from Mycobacterium tuberculosis (MVA85A) or recombinant hepatitis B virus surface antigen (rHBsAg) vaccines. The significant adjuvant activity observed provides good evidence supporting our hypothesis that CCR4 is a viable target for rational adjuvant design.     

CD4+CD25+T细胞与支气管哮喘

本文就CD4^＋CD25^＋T细胞的调节机制，CD4^＋CD25^＋T细胞与变态反应的关系，以及CD4^＋CD25^＋T细胞在支气管哮喘发病机制中的作用作一综述。     

地塞米松对哮喘小鼠CD4+CD25+调节性T细胞及IL-4、IL-10水平的影响

目的 探讨地塞米松对哮喘小鼠CD4+ CD25+调节性T细胞及IL-4、IL-10水平的影响.方法 30只雄性BALB/c小鼠随机分为三组:正常对照组、哮喘组和地塞米松组.利用卵清白蛋白腹腔注射和雾化吸入制备哮喘模型;通过流式细胞仪检测各组小鼠脾脏单个核细胞CD4+ CD25+调节性T细胞占CD4+T细胞的百分比;使用免疫组织化学方法分析各组IL-4在小鼠肺组织中的表达情况;用酶联免疫吸附试验检测各组小鼠血清IL-10的水平.结果 哮喘组脾脏单个核细胞CD4+CD25+调节性T细胞百分比及IL-10的表达水平较正常对照组和地塞米松组降低(P＜0.05),哮喘组IL-4水平较正常对照组和地塞米松组增高(P＜0.05).结论 地塞米松的抗炎作用可能通过上调CD4+ CD25+调节性T细胞、调节性T细胞亚群失衡的途径来实现.     

Artificial antigen-presenting cells plus IL-15 and IL-21 efficiently induce melanoma-specific cytotoxic CD8+ CD28+ T lymphocyte responses

ObjectiveTo develop a novel artificial antigen-presenting system for efficiently inducing melanoma-specific CD8⁺ CD28⁺ cytotoxic T lymphocyte (CTL) responses.MethodsCell-sized Dynabeads® M-450 Epoxy beads coated with H-2K^b: Ig-TRP2_180-188and anti-CD28 antibody were used as artificial antigen-presenting cells (aAPCs) to induce melanoma-specific CD8⁺CD28⁺ CTL responses with the help of IL-21 and IL-15. Dimer staining, proliferation, ELISPOT, and cytotoxicity experiments were conducted to evaluate the frequency and activity of induced CTLs.ResultsDimer staining demonstrated that the new artificial antigen-presenting system efficiently induced melanoma TRP2-specific CD8⁺CD28⁺CTLs. Proliferation and ELISPOT assays indicated that the induced CTLs rapidly proliferate and produce increased IFN- γ under the stimulation of H-2K^b: Ig-TRP2-aAPCs, IL-15, and IL-21. In addition, cytotoxicity experiments showed that induced CTLs have specific killing activity of target cells.ConclusionsThe new artificial antigen-presenting system including aAPCs plus IL-21 and IL-15 can induce a large number of antigen-specific CD8⁺ CD28⁺ CTLs against the melanoma. Our study provides evidence for a novel adoptive immunotherapy against tumors.     

Discordant memory B cell and circulating anti-Env antibody responses in HIV-1 infection

Long-lived memory B cells (B_Mem) provide an archive of historic Ab responses. By contrast, circulating Abs typically decline once the immunogen is cleared. Consequently, circulating Abs can underestimate the nature of cognate humoral immunity. On the other hand, the B_Mem pool should provide a comprehensive picture of Ab specificities that arise over the entire course of infection. To test this hypothesis, we compared circulating Ab and B_Mem from natural virus suppressors who control HIV-1 without therapy and maintain a relatively intact immune system. We found high frequencies of B_Mem specific for the conserved neutralizing CD4 induced or CD4 binding site epitopes of gp120, whereas low Ab titers to these determinants were detected in contemporaneous plasma. These data suggest that plasma Ab repertoires can underestimate the breadth of humoral immunity, and analyses of B_Mem should be included in studies correlating Ab specificity with protective immunity to HIV-1.     

CD4+CXCR5+ T cells activate CD27+IgG+ B cells via IL-21 in patients with hepatitis C virus infection

BACKGROUND: Chronic hepatitis C virus (HCV) infection causes the skewing and activation of B cell subsets, but the characteristics of IgG+ B cells in patients with chronic hepa-titis C (CHC) infection have not been thoroughly elucidated. CD4+CXCR5+ follicularhelperT(Tfh)cells,viainterleukin (IL)-21 secretion, activate B cells. However, the role of CD4+CXCR5+T cellsintheactivationof IgG+ BcellsinCHCpatientsis not clear.
METHODS: The frequency of IgG+ B cells, including CD27?IgG+B and CD27+IgG+ B cells,the expression of the activation markers (CD86 and CD95) in IgG+ B cells, and the percentage of circu-lating CD4+CXCR5+ T cells were detected by flow cytometry in CHC patients (n=70) and healthy controls (n=25). The con-centrations of serum IL-21 were analyzed using ELISA. The role of CD4+CXCR5+ T cells in the activation of IgG+ B cells was investigated using a co-culture system.
RESULTS: A significantly lower proportion of CD27+IgG+ B cells with increased expression of CD86 and CD95 was observed in CHC patients.The expression of CD95 was negatively correlated with the percentage of CD27+IgG+ B cells, and it contributed to CD27+IgG+ B cell apoptosis. Circulating CD4+CXCR5+ T cells and serum IL-21 were significantly increased in CHC patients. Moreover, circulating CD4+CXCR5+ T cells from CHC patients induced higher expressions of CD86 and CD95 in CD27+IgG+B cells in a co-culture system; the blockade of the IL-21 decreased the expression levels of CD86 and CD95 in CD27+IgG+ B cells.
CONCLUSIONS: HCV infection increased the frequency of CD4+CXCR5+ T cells and decreased the frequency of CD27+IgG+B cells. CD4+CXCR5+ T cells activated CD27+IgG+ B cells via the secretion of IL-21.     

CD4+CD25+调节性T细胞与支气管哮喘

支气管哮喘是一种常见的慢性呼吸道疾病,其免疫发病机制尚不十分清楚。CD4 CD25 调节性T细胞是一种特殊的调节性T细胞,参与自身免疫调节,维持自身免疫耐受。本文就CD4 CD25 调节性T细胞的特性及与支气管哮喘的发病机制、治疗、预后的研究进展做一综述。     

支气管哮喘大鼠CD4^＋CD25^＋T细胞的变化及地塞米松干预作用的研究

目的研究支气管哮喘（简称哮喘）大鼠模型支气管肺泡灌洗液（BALF）、血液、脾脏CD4^＋CD25^＋T细胞的变化,及地塞米松对CD4^＋CD25^＋T细胞的影响。方法50只SD大鼠随机分为5组,空白对照（A）组,哮喘（B）组,地塞米松1（C）组、地塞米松2（D）组,地塞米松3（E）组。A组第1天给予腹腔注射生理盐水1ml,第15～21天每天给予生理盐水雾化。B、C、D、E组用卵蛋白建立哮喘大鼠模型,第1天,每只大鼠腹腔注射抗原1ml（卵蛋白1mg＋灭活百日咳杆菌9×10。个＋氢氧化铝干粉100mg）混悬液,第15～21天给予1％的卵蛋白雾化30min,C、D、E组于雾化后分别给予腹腔注射地塞米松0．2mg／kg、1mg／kg、2mg／kg。采用流式细胞仪检测的方法,观察大鼠体内BALF、外周血、脾脏CD4^＋CD25^＋T细胞的变化及使用不同剂量地塞米松后对其的影响。结果B组BALF、外周血、脾脏CD4^＋CD25^＋T细胞表达占CD4^＋T细胞的百分比分别是（42．21±5．62）％、（12．69±2．70）％、（11．15±1．05）％,A组结果分别是（18．76±5．85）％、（6．21±1．73）％、（7．85±2．13）％。B组与A组比较,差异均具有统计学意义（P〈0．01,P〈0．01,P〈0．05）;C组、D组、E组BALF中CD4^＋CD25^＋T细胞占CD4^＋T细胞的百分比表达分别是（10．49±4．03）oA、（13．28±5．12）％、（7．51±5．39）％,显著低于A组和B组,（P〈0．05,P〈0．01）;外周血中,C组（6．03±1．43）％、D组（4．88±0．95）％与A组（6．21±1．73）％比较,差异无统计学意义,E组（3．49士0．62）％与C组、A组比较,差异有统计学意义（P〈0．05）。脾脏中,c组（7．25±1．82）%、D组（8．63±3．18）％与A组（7．85±2．13）％比较,差异无统计学意义,E组（3．38±1．37）％与C组、D组、A组比较,差异有统计学意义（P〈0．05）。结论CD4^＋CD25^＋T细胞在哮喘大鼠体内有明显的优势表达,可能是哮喘发病的机制之一。地塞米松可以抑制CD4^＋CD25^＋T细胞的表达。BALF内CD4^＋CD25^＋T细胞的变化与外周血和脾脏的变化具有一致性,监测外周血或脾脏CD4^＋CD25^＋T细胞变化可了解肺部情况。     

CD4+ T cells in lung tissue predict responsiveness to cyclosporin A in interstitial pneumonia

OBJECTIVE: A limited number of case studies have demonstrated the steroid-sparing and disease stabilization effects of cyclosporin A (CsA) combined with corticosteroid in patients with chronic interstitial pneumonia (IP). Although CsA is known to inhibit the proliferation and function of CD4+ T cells, it is not clear what type of IP is responsive to CsA administration. METHODOLOGY: In order to evaluate whether any lymphocyte subsets in alveolar tissue predict the responsiveness to CsA, morphometric analysis was performed on lung tissue obtained from six IP patients who were treated with a combination of CsA and steroid because the therapeutic effect of steroid alone had been limited. RESULTS: Cell densities of CD4+ T cells were significantly decreased in the alveolar tissues obtained from CsA responders (n = 3) compared to non-responders (n = 3) (120 +/- 86/mm2 vs 503 +/- 172/mm2, P=0.0495). CD4/CD8 ratios of CsA responders were also significantly lower than those of non-responders (0.315 +/- 0.070 vs 1.975 +/- 0.965, P=0.0463). However, cell densities of CD8+ T cell and areas of B cells aggregates were similar in both groups. CONCLUSIONS: The present observations, contrary to expectation, reveal that lower cell densities of CD4+ T cells and lower CD4/CD8 ratios are associated with responsiveness to CsA in combination therapy, suggesting that pharmacological actions other than suppression of CD4+ T cells explain the efficacy of CsA in patients with chronic IP.     

支气管哮喘大鼠CD4+CD25+T细胞的变化及地塞米松干预作用的研究

目的 研究支气管哮喘(简称哮喘)大鼠模型支气管肺泡灌洗液(BALF)、血液、脾脏CD4+CD25+T细胞的变化,及地塞米松对CD4+CD25+T细胞的影响.方法 50只SD大鼠随机分为5组,空白对照(A)组,哮喘(B)组,地塞米松1(C)组、地塞米松2(D)组,地塞米松3(E)组.A组第l天给予腹腔注射生理盐水l ml,第15～21天每天给予生理盐水雾化.B、C、D、E组用卵蛋白建立哮喘大鼠模型,第1天,每只大鼠腹腔注射抗原l ml(卵蛋白1 mg+灭活百日咳杆菌9×106个+氢氧化铝干粉100 mg)混悬液,第15～21天给予1%的卵蛋白雾化30 min,C、D、E组于雾化后分别给予腹腔注射地塞米松0.2 mg/kg、1 mg/kg、2 mg/kg.采用流式细胞仪检测的方法 ,观察大鼠体内BALF、外周血、脾脏CD4+CD25+T细胞的变化及使用不同剂量地塞米松后对其的影响.结果 B组BALF、外周血、脾脏CD4+CD25+T细胞表达占CD4+T细胞的百分比分别是(42.21±5.62)%、(12.69±2.70)%、(11.15±1.05)%,A组结果 分别是(18.76±5.85)%、(6.21±1.73)%、(7.85±2.13)%.B组与A组比较,差异均具有统计学意义(P＜0.01,P＜0.01,P＜0.05);C组、D组、E组BALF中CD4+CD25+T细胞占CD4+T细胞的百分比表达分别是(10.49±4.03)%、(13.28±5.12)%、(7.51±5.39)%,显著低于A组和B组,(P＜0.05,P＜0.01);外周血中,C组(6.03±1.43)%、D组(4.88±0.95)%与A组(6.21±1.73)%比较,差异无统计学意义,E组(3.49±0.62)%与C组、A组比较,差异有统计学意义(P＜0.05).脾脏中,C组(7.25±1.82)%、D组(8.63±3.18)%与A组(7.85±2.13)%比较,差异无统计学意义,E组(3.38±1.37)%与C组、D组、A组比较,差异有统计学意义(P＜0.05).结论 CD4+CD25+T细胞在哮喘大鼠体内有明显的优势表达,可能是哮喘发病的机制之一.地塞米松可以抑制CD4+CD25+T细胞的表达.BALF内CD4+CD25+T细胞的变化与外周血和脾脏的变化具有一致性,监测外周血或脾脏CD4+CD25+T细胞变化可了解肺部情况.     

CD4+ T cells mediate mucosal and systemic immune responses to experimental hookworm infection

Hookworm infection is associated with anaemia and malnutrition in many resource‐limited countries. Ancylostoma hookworms have previously been shown to modulate host cellular immune responses through multiple mechanisms, including reduced mitogen‐mediated lymphocyte proliferation, impaired antigen presentation/processing, and relative reductions in CD4⁺ T cells in the spleen and mesenteric lymph nodes. Syrian hamsters were depleted of CD4⁺ for up to 9 days following intraperitoneal injection (200 μg) of a murine anti‐mouse CD4 monoclonal IgG (clone GK1·5). CD4⁺ T‐cell‐depleted hamsters infected with the hookworm Ancylostoma ceylanicum exhibited a threefold higher mean intestinal worm burden and more severe anaemia than animals that received isotype control IgG. In addition, depletion of CD4⁺ T cells was associated with impaired cellular and humoral (serum and mucosal) immune responses to hookworm antigens. These data demonstrate an effector role for CD4⁺ T cells in hookworm immunity and disease pathogenesis. Ultimately, these studies may yield important insights into the relationship between intestinal nematode infections and diseases that are associated with CD4⁺ T‐cell depletion, including HIV.     

肺结核患者外周血CD_4~+、CD_8~+T细胞Blimp-1表达下降

目的研究活动性肺结核患者外周血单个核细胞(PBMCs)Blimp-1的表达及临床意义。方法采集31例活动期肺结核患者和45位健康对照组外周血,纯化PBMCs,用结核分枝杆菌ESAT-6和CFP-10混合性抗原肽库刺激,通过细胞表面标记和细胞内细胞因子染色技术,采用流式技术检测CD+4、CD+8T细胞Blimp-1的表达。结果与对照组比较,肺结核患者PBMCs中的CD+4、CD+8T细胞亚群分布出现显著性下降,且肺结核患者CD+4T细胞中Blimp-1的表达比例(%)下降(肺结核组89.5%(83.8%,95.7%),对照组94.5%(89.8%,98.7%),P0.05),且CD+4、CD+8T细胞中Blimp-1的表达量(平均荧光强度)也显著性下降(CD+4T细胞:肺结核组9.28(7.5,18.9),对照组15.4(11,25.4),P0.05);CD+8T细胞:肺结核组9.01(6.08,14.7),对照组14.2(9.53,23.1),P0.05)。结论活动期肺结核CD+4、CD+8T细胞群内Blimp-1的表达下降可能会使效应性和调节性T细胞的分化出现异常。Blimp-1可能参与结核病的疾病进程,这为研究结核病的诊断和治疗提供了线索。     

结核性胸膜炎患者胸液中CD4~+ CD25~+ FoxP3~+调节T细胞抑制抗结核免疫

目的研究结核性胸膜炎患者胸液中CD4+CD25+FoxP3+调节T细胞是否增多,这些调节T细胞是否抑制结核的特异细胞免疫反应。方法使用细胞分离、流式细胞分析及体外细胞培养作细胞增殖及增殖抑制等实验方法,对15例结核性胸膜炎患者及17例健康正常人群胸液及外周血白细胞中CD4+CD25+FoxP3+调节T细胞的量及特征作研究。结果结核性胸膜炎患者胸液中CD4+CD25+FoxP3+调节T细胞明显高于患者及健康人群外周血。在体外,结核性胸膜炎患者胸液中单核细胞对BCG刺激产生γ-干扰素(IFN-γ)的能力明显强于患者及健康人群外周血中单核细胞;把这些调节T细胞从胸液单核细胞中清除,增强了结核患者胸液单核细胞对BCG刺激产生IFN-γ;从结核患者胸液分离的这些调节T细胞能抑制结核患者Th1细胞产生IFN-γ。结论结核性胸膜炎患者胸液CD4+CD25+FoxP3+调节T细胞增多,抑制结核性胸膜炎患者Th1细胞免疫反应,从而参与了结核性胸膜炎的发病。     

Priming of CD4+ T cells and development of CD4+ T cell memory; lessons for malaria

CD4 T cells play a central role in the immune response to malaria. They are required to help B cells produce the antibody that is essential for parasite clearance. They also produce cytokines that amplify the phagocytic and parasitocidal response of the innate immune system, as well as dampening this response later on to limit immunopathology. Therefore, understanding the mechanisms by which T helper cells are activated and the requirements for development of specific, and effective, T cell memory and immunity is essential in the quest for a malaria vaccine. In this paper on the CD4 session of the Immunology of Malaria Infections meeting, we summarize discussions of CD4 cell priming and memory in malaria and in vaccination and outline critical future lines of investigation. B. Stockinger and M.K. Jenkins proposed cutting edge experimental systems to study basic T cell biology in malaria. Critical parameters in T cell activation include the cell types involved, the route of infection and the timing and location and cell types involved in antigen presentation. A new generation of vaccines that induce CD4 T cell activation and memory are being developed with new adjuvants. Studies of T cell memory focus on differentiation and factors involved in maintenance of antigen specific T cells and control of the size of that population. To improve detection of T cell memory in the field, efforts will have to be made to distinguish antigen-specific responses from cytokine driven responses.     

CD4+CD25+调节性T细胞及其与移植物抗宿主病和移植物抗白血病的关系

CD4 CD25 调节性T细胞(Treg细胞)是CD4 T细胞的一个亚群,在维持机体自身免疫耐受,诱导移植耐受等方面发挥重要作用。移植物抗宿主病(GVHD)是异基因造血干细胞移植最严重的并发症之一。在动物骨髓移植模型中证实Treg细胞可以促进移植物植入,减少GVHD的发生率和严重度,但并没有消除移植物抗白血病(GVL)的作用。在人体有关Treg细胞对GVHD的影响因研究者分析Treg细胞采用的表型不同,其结论存在不一致性。本文就新近Treg细胞生物学特性,Treg细胞与GVHD和GVL的关系的研究进展进行综述。     

Increased frequency of CD4+PD-1+HLA-DR+ T cells is associated with disease progression in CLL

Chronic lymphocytic leukaemia (CLL) patients often have abnormal expansions of CD4⁺ and CD8⁺ T cells and this can be associated with progressive disease. To characterise the key T-cell populations involved in this phenomenon, we used flow cytometry and 11 phenotypic markers to study 74 CLL patients and 14 controls. T cells of CLL patients were more phenotypically complex than those of healthy controls with significant increases in the frequencies of CD4 and CD8 memory T cells expressing exhaustion-, activation- and senescence-associated markers. Multivariate analysis of 111 different T-cell subsets showed that high frequencies of four subsets (three CD8 and one CD4) were associated with shorter progression-free survival. The most significant association was with CD4⁺HLA-DR⁺PD-1⁺ T cells, and patients could be stratified into high- and low-risk groups based on the frequency of these T cells. The expansion of this CD4⁺ subset could not be accounted for by age, cytomegalovirus infection or increases in Treg cells. Overall, these results highlight two relatively simple biomarkers, percentage CD8⁺ and percentage CD4⁺PD-1⁺HLA-DR⁺ T cells, which can be used to risk-stratify CLL patients, independent of other tumour-associated markers. They also provide further evidence for the pivotal role of T cells in modulating the pathology of CLL.