Molecular aspects of regulatory T cell development

Naturally arising CD4(+) regulatory T cells are critical mediators of peripheral self tolerance and immune homeostasis. Emerging evidence suggests that these cells are a dedicated T cell lineage constitutively acting in a dominant, cell extrinsic manner to regulate immune function. These observations are driving a reevaluation of our understanding of the regulation of immune homeostasis and peripheral tolerance. But despite a growing body of phenomenological data, an understanding of the molecular mechanisms underlying CD4(+) regulatory T cell development has only recently begun to emerge. This review highlights the major molecular pathways that have been implicated to date in the development of this T cell lineage.     

Th17 and regulatory T cell balance in autoimmune and inflammatory diseases

This review focuses on the biology of T helper 17 (Th17) and regulatory T (Treg) cells and their role in inflammatory diseases, such as rheumatoid arthritis. Th17 cells represent a pro-inflammatory subset whereas Treg cells have an antagonist effect. Their developmental pathways are reciprocally interconnected and there is an important plasticity between Th17 and Treg cells. These features implicate that the Th17/Treg balance plays a major role in the development and the disease outcomes of animal model and human autoimmune/inflammatory diseases. During these diseases, this balance is disturbed and this promotes the maintenance of inflammation. Targeting the Th17/Treg imbalance can be performed at different levels such as inhibition of pro-inflammatory cytokines and their receptors, of pathogenic cells or their specific signaling pathways. Conversely, direct effects include administration or induction of protective cells, or stimulation of their specific pathways. Several clinical trials are underway and some positive results have been obtained.     

Composition and plasticity of triple-negative breast carcinoma-infiltrating regulatory T cells

Low Foxp3⁺ regulatory T-cell (Treg) presence in the tumor-infiltrating lymphocytes (TILs) is considered favorable in breast cancer, and numerous CD25-targeting agents have been applied in the attempt to remove Foxp3⁺ Treg cells, which typically present CD4⁺CD25^+/hi surface phenotype. However, CD25 is not Treg-exclusive and can be upregulated by effector T cells. Hence, CD25 depletion may cause the elimination of activated T cells that are responding to tumor-specific antigens. In this study, the composition and function of CD4⁺CD25⁺ cells inside the microenvironment of triple-negative breast carcinoma (TNBC) were investigated. Directly ex vivo, the Foxp3⁺ Treg cells represented a minor subset in total CD4⁺CD25⁺ TILs. Significant differences were observed in the expression of Treg-associated molecules between CD4⁺CD25⁺Foxp3⁺ TILs and CD4⁺CD25⁺Foxp3⁻ TILs. While both the CD4⁺CD25⁺Foxp3⁺ and the CD4⁺CD25⁺Foxp3⁻ TILs could express CTLA-4 and LAG-3, the expression levels were significantly higher in CD4⁺CD25⁺Foxp3⁺ TILs than in CD4⁺CD25⁺Foxp3⁻ TILs. Upon TCR stimulation, the expression of TGF-beta was significantly higher in CD4⁺CD25⁺Foxp3⁺ TILs, while the expression of IL-10 was significantly higher in CD4⁺CD25⁺Foxp3⁻ TILs. These differences were conserved in the blood counterparts of these cells. Interestingly, the level of CD25⁺Foxp3⁺ cells in circulating CD4⁺ T cells was positively correlated with the level of CD25⁺Foxp3⁺ cells in CD4⁺ TILs, but the level of CD25⁺Foxp3⁻ cells in circulating CD4⁺ T cells was not associated with the level of CD25⁺Foxp3^- cells in CD4⁺ TILs. Th17-polarizing medium could readily remodel CD4⁺CD25⁺Foxp3⁻, but not CD4⁺CD25⁺Foxp3⁺, T cells into RORgammat and IL-17-expressing T cells, demonstrating stronger plasticity of the former subset. Together, these data demonstrated that the CD4⁺CD25⁺ TILs were composed of distinctive Foxp3⁻ and Foxp3⁺ cells, with the former representing the major subset. The antigen specificity and effector molecule expression of the CD4⁺CD25⁺Foxp3⁻ thus require further analyses.     

Involvement of T helper type 17 and regulatory T cell activity in Citrobacter rodentium invasion and inflammatory damage

Citrobacter rodentium is a murine pathogen that transiently colonizes the lumen of the large intestine. C. rodentium induces colitis, but the relative importance and temporal induction of the T helper type 17 (Th17) and regulatory T cell (T_reg) pathways in protection from the infection and inflammation have not been assessed. Our aim was to investigate the key immunological signalling events associated with successful clearance of C. rodentium. Mice were challenged with luminescent-tagged C. rodentium and killed at days 3 (early infection), 10 (peak infection) and 21 (late infection) post-infection. Bioluminescent imaging and bacterial culture determined levels of C. rodentium. Distal colon mRNA expression of interleukin (IL)-17, IL-6, IL-1β, tumour necrosis factor (TNF)-α, forkhead box P3 (FoxP3) and ghrelin were assessed using real-time polymerase chain reaction. Results were compared with age-matched non-infected mice. Low levels of C. rodentium were found at day 3, high levels at day 10, with clearance from the majority of the mice by day 21. In the distal colon, there was up-regulation of TNF-α and FoxP3 throughout the study and increases in IL-6 and IL-17 during the peak and late stages of infection. Ghrelin expression was increased at the peak and late stages of infection. This study has characterized changes to the T helper cell pathways, following the course of C. rodentium infection in mice. There were significant immunological changes, with up-regulation of the Th17 and T_reg pathways in the distal colon and an increase in ghrelin expression compared with non-infected control mice. These changes may play a role in the pathology and clearance of C. rodentium.     

Mechanisms of T regulatory cell function

Regulatory T cells (Treg) play a pivotal role in tolerance to self-antigens and tissue grafts, and suppression of autoimmune reactions. These cells modulate the intensity and quality of immune reactions through attenuation of the cytolytic activities of reactive immune cells. Treg cells operate primarily at the site of inflammation where they modulate the immune reaction through three major mechanisms: a) direct killing of cytotoxic cells through cell-to-cell contact, b) inhibition of cytokine production by cytotoxic cells, in particular interleukin-2, c) direct secretion of immunomodulatory cytokines, in particular TGF-beta and interleukin-10. In addition to differential contributions of these mechanisms under variable inflammatory conditions, mechanistic complexity and diversity evolves from the diverse tasks performed by various Treg cell subsets in different stages of the immune reaction. Here we attempt to integrate the current experimental evidence to delineate the major suppressive pathways of Treg cells.     

树突状细胞对调节性T细胞的调控作用

树突状细胞(DCs)是目前已知的体内功能最强大的专职性抗原提呈细胞,具有启动免疫应答和诱导免疫耐受的双重特性.近年来树突状细胞对调节性T细胞的调控作用是免疫学领域的一个研究热点.Foxp3+ Tregs是一群同时具有免疫低反应性和免疫抑制性功能两大特征的T淋巴细胞,它在维持机体内环境稳定、预防自身免疫性疾病、抑制移植排斥反应等病理生理过程中发挥着重要作用.越来越多的研究结果证实DCs和Tregs二者在维持外周免疫耐受中存在着紧密联系,DCs可以诱导抗原特异性Tregs的生成并增加后者的抑制活性,其中参与该调节机制的分子主要包括相关细胞因子、Toll样受体、共刺激分子及维甲酸等.对DCs在接触共生和致病微生物时诱导和调控Tregs细胞有了一些新发现.     

The regulatory, inflammatory, and T cell programming roles of interleukin-2 (IL-2)

Signaling through IL-2 induces the activation of pathways that lead to the proliferation, survival and cytokine production of effector T cells. However, through negative feedback mechanisms, internalization of the IL-2 receptor, induction of activation-induced cell death, and the generation of regulatory T cells, IL-2 also promotes the suppression of inflammatory responses. In regulatory T cells, IL-2 signaling upregulates the expression of FoxP3. Regulatory T cell induction by TGF-beta also requires IL-2. Additionally, pro-inflammatory and pro-survival pathways involving PI3K upon IL-2 stimulation is inhibited by PTEN in regulatory T cells. Importantly, IL-2 signaling is key for the development, expansion and maintenance of regulatory T cells. However, gamma(c) cytokines can replace requirements for IL-2 in regulatory T cells, although not with the same efficacy. The dual roles of IL-2 in inflammation are demonstrated in that mice deficient in both FoxP3 and IL-2 display less severe symptoms compared to FoxP3 deficient mice. Finally, IL-2 not only plays a key role in the induction of effector T cells and regulatory T cells, it also inhibits IL-17 producing T cells. By understanding complex dynamics of IL-2 interactions in the inflammatory response, therapies may be developed or modified for regulating immune related diseases.     

Special regulatory T cell review: How I became a T suppressor/regulatory cell maven

I have briefly reviewed the factors that motivated me to change my views about the existence and importance of suppressor/regulatory T cells and to devote the majority of my laboratory efforts to this newly revitalized area of immunologic research. I am optimistic that manipulation of regulatory T-cell function will shortly be applicable to the clinic.     

Impairment of T helper and T regulatory cell responses at birth

Background: There is strong evidence that reduced exposures to microbial compounds triggering innate immune responses early in life are critical for the development of allergic illnesses. The underlying mechanisms remain unknown, but will include T‐cell responses either along T helper type 1 (Th1)/Th2 pathways or via T regulatory and Th17 cells. Yet, little is known about innate immune responses and the function of T regulatory/Th17 cells at birth. The aim of this study was to investigate T‐cell responses to innate (Lipid A/LpA, peptidoglycan/Ppg) and adaptive (phytohemagglutinin) stimuli at birth and to compare these findings with adult immune responses. Methods: Cord and peripheral blood mononuclear cells including T regulatory and Th17 cells from 25 neonates and 25 adults were examined for proliferation, cytokine secretion, surface, mRNA expression and functional suppression assays. Results: Proliferation and cytokine responses to innate stimuli were less mature at birth than in adulthood. T regulatory and Th17 cells were less expressed in cord than in adult blood (Ppg‐induced Foxp3, P = 0.001, LpA‐induced CD4⁺ CD25⁺ high, P = 0.02; Th17 : P < 0.0001). Mitogen‐induced suppression of T‐regulatory cells on T‐effector cell function was less efficient in cord than in adult blood (P = 0.01). At both ages, Th17 cells were correlated with Th1/Th2 cells (P < 0.01), but not with interleukin‐10 secretion following innate‐stimulation. Conclusion: Innate immune responses are immature at birth. Furthermore, the function of T regulatory and Th17 cells is impaired. Th17 cells in association with Th1/Th2 cells may be involved in early immuno‐modulation. Potent innate immune stimulation early in life can potentially contribute to protection from allergic diseases.     

小鼠派氏集合淋巴结T细胞活化和调节性T细胞的分析

目的： 通过对小鼠派氏集合淋巴结（PPs）、肠系膜淋巴结（MLNs）和腹股沟淋巴结（ILNs）的比较性研究，以分析PPs T细胞活化和调节性T细胞的功能特点。 方法： 无菌分离小鼠PPs、MLNs和ILNs，分别制备单个淋巴细胞悬液，运用流式细胞术结合荧光抗体染色技术来检测CD3+T细胞、CD3+CD4+辅助性T细胞和CD4+CD25＋调节性T细胞的比例；用多克隆刺激剂刀豆蛋白A（Con A）、佛波醇酯（PDB）和离子霉素（Ion） 刺激活化淋巴细胞，随后运用流式细胞术结合双色荧光抗体染色技术来检测T细胞早期活化标志CD69的表达水平。 结果： PPs内CD3+T细胞所占比例明显低于MLNs和ILNs，然而CD4+CD25＋调节性T细胞的比例明显高于MLNs和ILNs；在没有加入刺激剂的培养条件下，PPs来源的T细胞CD69的表达水平明显高于MLNs和ILNs来源的T细胞，MLNs来源的T细胞CD69的表达水平明显高于ILNs来源的T细胞；而在加入Con A或者单纯PDB的培养条件下，PPs来源的T细胞却表现出低反应性；在加入PDB＋Ion的培养条件下，3者来源的T细胞CD69的表达水平没有明显差别。 结论： PPs内CD3+T细胞所占比例较低，CD4+CD25＋调节性T细胞的比例较高，这可能是PPs整体T细胞低反应性的原因之一；PPs来源T细胞的高基础活化率可能与其不断接触小肠来源的食物和共生菌抗原有关；PPs来源T细胞选择性地对某些刺激剂表现出低反应性，提示这群细胞处于某种程度的无能状态。上述特征的生物学意义在于避免因为不断接受小肠来源的食物和共生菌抗原的刺激而发生病理性炎症的同时还保留对病原微生物抗原的应答。