Treg Differentiation Research Articles

STING is an intrinsic checkpoint inhibitor that restrains the TH17 cell pathogenic program.

SUMMARYExternal and intrinsic factors regulate the transcriptional profile of T helper 17 (TH17) cells, thereby affecting their pathogenic potential and revealing their context-dependent plasticity. The stimulator of interferon genes (STING), a component of the intracellular DNA-sensing pathway, triggers immune responses but remains largely unexplored in T cells. Here, we describe an intrinsic role of STING in limiting the TH17 cell pathogenic program. We demonstrate that non-pathogenic TH17 cells express higher levels of STING than those activated under pathogenic conditions. Activation of STING induces interleukin-10 (IL-10) production in TH17 cells, decreasing IL-17A and IL-23R expression in a type I interferon (IFN)-independent manner. Mechanistically, STING-induced IL-10 production partially requires aryl hydrocarbon receptor (AhR) signaling, while the decrease of IL-17A expression occurs due to a reduction of Rorγt transcriptional activity. Our findings reveal a regulatory function of STING in the TH17 cell activation program, proposing it as a valuable target to limit TH17-cell-mediated inflammation.

ROCK2 Specific Inhibition Attenuates DOCA Salt‐Induced Cardiac Fibrosis and Renal T Cell Infiltration

Hypertension, or an elevated blood pressure, is the primary modifiable risk factor for cardiovascular disease, the number one cause of mortality worldwide. We previously demonstrated that Th17 activation and interleukin 17A (IL‐17A)/IL‐21 production is integral for the full development of a hypertensive phenotype as well as the renal and vascular damage associated with hypertension. Rho‐associated coiled‐coil containing protein kinase 2 (ROCK2) serves as a molecular switch upregulating Th17 and inhibiting regulatory T cell (Treg) differentiation of CD4+ T cells. Initially, we measured ROCK2 expression and found that ROCK2 is upregulated in splenic and renal CD4+ T cells in hypertensive mice, confirming the potential for influencing hypertension‐associated T cell activation. We then hypothesized that excessive T cell ROCK2 activation, leading to increased Th17/Treg ratios, contributes to hypertension and ultimately, end‐organ damage. We first showed in vitro that KD025, an orally bioavailable ROCK2 inhibitor, inhibits Th17 proliferation and IL‐17A/IL‐21 production. To determine if hypertensive stimuli such as endothelial stretch increase T cell ROCK2 expression, we cultured human aortic endothelial cells exposed to 5% (normotensive) or 10% (hypertensive) stretch with circulating human T cells and HLA‐DR+ antigen presenting cells. Hypertensive stretch increased T cell ROCK2 expression 2‐fold. We then tested the effect of ROCK2 inhibition in vivo on DOCA (deoxycorticosterone) salt‐induced hypertension. DOCA‐salt (uninephrectomy + 100mg DOCA pellet + 1% NaCl drinking water, n=19) was initiated in all animals allowing for hypertension to develop for 10 days, followed by treatment with the ROCK2 specific inhibitor KD025 (50mg/kg i.p. daily starting on Day 11, n=11) for the remainder of the 21‐day protocol. KD025 treatment significantly attenuated cardiac hypertrophy (8.7±0.29 vs 7.6±0.21 mg/grams body weight, p=0.005) and left ventricular (LV) fibrosis (4.9±1.7 vs 2.4±1.3% LV area, p=0.008) by the termination of the study. Flow cytometric analysis revealed that KD025 treatment attenuated renal leukocyte infiltration (414.6±81.5 vs 317.1±43.6 cells/mg tissue, p=0.057) primarily due to an attenuation in the number of CD3+ T cell infiltrates (58.99±5.55 vs 43.45±3.77 cells/mg tissue, p=0.019). Additional studies were performed using the Angiotensin II (Ang II, 490ng/kg/min) model of hypertension which demonstrated that KD025 treatment starting on Day 1 of Ang II infusion significantly blunted the hypertensive response as early as one week of Ang II treatment (139.4±8.5 vs 108±4.8 mmHg systolic blood pressure, p<0.05, n=5‐6). This protection was accompanied by a decreased Th17/Treg ratio (1.66±0.11 vs 1.31±0.13, p=0.029) of renal T cells in vivo, confirming the effects of KD025 observed in the prior in vitro experiments. These data indicate that the ROCK2‐specific inhibitor KD025 may be a novel therapeutic for the treatment of hypertension and the associated tissue damage.

Lymph node-targeted long-acting butyrate micelles induce regulatory immune modulation

Abstract Regulatory T cells (Tregs) play essential roles in maintaining immune homeostasis and preventing autoimmunity and allergy. Peripherally derived Tregs are induced most efficiently in the lymph nodes (LNs), yet current therapies are not effectively delivered to these sites. The short-chain fatty acid butyrate facilitates extrathymic differentiation of Tregs and has been considered a promising therapeutic candidate. However, there are challenges to its use, including its foul odor, quick metabolism, and low efficiency with traditional oral delivery. Herein, we designed butyrate-prodrug micelles that deliver butyrate to peripheral LNs through subcutaneous (SC) administration. We have developed two polymeric micelles, one with a neutral charge (termed NtL-ButM) and one with a negative charge (termed Neg-ButM), which both contain 28 wt% butyrate and have similar sizes of ~40 nm. In a biodistribution study, we observed that Neg-ButM, not NtL-ButM, accumulated in draining LNs for over one month. At the cellular level, the Neg-ButM were taken up mostly by the macrophages and dendritic cells in the LNs and inhibited LPS-induced activation of those cells. In antibiotic-treated mice, three weekly doses of Neg-ButM significantly increased the numbers of CD25+ Foxp3+ CD4+ Tregs in draining LNs compared to either PBS or NtL-ButM treatment. Furthermore, we demonstrated that, when combined with intragastrically administered peanut proteins, the SC-injected Neg-ButM protected from anaphylactic responses in a mouse model of peanut allergy. This approach provides a potential Treg-based therapy against food allergies. Supported by a seed grant from the Chicago Immunoengineering Innovation Center of the University of Chicago.

Interleukin-2 signaling engages de novo cholesterol biosynthesis in regulatory T cells

Abstract In the absence of IL-2R signaling, Treg homeostasis is severely disrupted. This is accompanied by a decrease in the expression of enzymes related to cholesterol biosynthesis, but the function of this pathway in Tregs is unknown. To investigate this, we selectively ablated Sterol Regulatory Binding Element Protein 2 by crossing Srebp2flox/flox mice to Foxp3cre mice (Srebp2cKO). Consistent with published studies, homozygous Srebp2cKO mice are outwardly normal. However, when Srebp2cKO Tregs are placed in competition with wild-type Tregs, they are found at a reduced frequency in the spleen, lymph nodes and gut mucosa. In this setting, Srebp2cKO Tregs have reduced Foxp3 expression, suggesting that their function and/or stability may be impacted by altered cholesterol biosynthesis. These defects persist in lymphoid and non-lymphoid tissues when Tregs in competition are treated with an IL-2R agonist. In addition, a marked reduction in effector Treg differentiation is associated with Srebp2cKO Tregs, as expression of CD39, CD73, CD103, Klrg1 and ICOS is reduced while Bcl-2 and CD62L are increased. In response to IL-2R signaling, Srebp2cKO Tregs do not optimally upregulate CD25, which suggests a function for cholesterol biosynthesis in supporting the high affinity IL-2R. These results are consistent with a model whereby IL-2R signaling coordinates Treg cholesterol biosynthesis, which contributes to effector Treg development and raises the possibility that cholesterol biosynthesis could be a target in the therapeutic manipulation of Tregs. Supported by grants from NIH (F30 AI157211-01A1, RO1 AI148675-01)

Inhibition of distinct glycolytic enzymes produces differential effects on CD4 T cell function

Abstract Background When T cells are activated, they upregulate glycolysis and take on a requisite Warburg phenotype. In this study, we evaluated the effect of inhibiting two distinct glycolytic enzymes, GAPDH and PGAM, on CD4 T cell differentiation and effector activities in order to gain a mechanistic understanding of the relationship between glycolysis and T cell function. Methods To evaluate the effect of GAPDH inhibition on T cell function, we cultured naïve murine CD4+ T cells under Th1 or Treg polarizing conditions and treated with either the GAPDH inhibitor heptelidic acid or the PGAM inhibitor EGCG. We then assessed either IFNγ production or identified CD25+Foxp3+ cells in response to either Th1 or Treg polarization, respectively. To evaluate the impact of GAPDH inhibition in vivo, we used the MOG35–55 experimental autoimmune encephalomyelitis (EAE) mouse model of multiple sclerosis (MS) and compared neurological function in mice treated with vehicle or heptelidic acid. Results We found that GAPDH inhibition had a potent anti-inflammatory effect on T cells in vitro, drastically reducing IFNγ production at both the transcriptional and protein levels. Interestingly, PGAM inhibition had the opposite effect, potently blocking Treg differentiation. In the EAE mouse model of MS, heptelidic acid attenuated the disease course, indicating that GAPDH inhibition is anti-inflammatory in vivo. Discussion GAPDH inhibition was anti-inflammatory in vitro and in vivo, while PGAM inhibition had the opposite effect. These findings suggest that the functional consequences of glycolysis inhibition depend on the specific enzymes targeted, which has both mechanistic and therapeutic implications for the role of glycolysis in immune cell function.

PD-1 restricts the development and effector function of tissue regulatory T cells in experimental autoimmune encephalomyelitis

Abstract PD-1 restricts T cell effector functions, regulates T cell tolerance, and maintains immune homeostasis, thereby playing pivotal roles in cancer, autoimmune and infectious diseases. Our studies and others show that PD-1 can restrain Treg suppressive function in autoimmunity and cancer. Here, we investigate the role of Treg intrinsic PD-1 in regulating pathogenic and protective CD4+ T cell responses in experimental autoimmune encephalomyelitis (EAE) by inducing PD-1 deletion on FoxP3+ cells prior to disease development. This deletion of PD-1 only in Treg protects mice from severe disease, and both flow cytometry and transcriptional analysis of central nervous system (CNS) CD4+ T cells show enrichment of tissue effector Treg (eTreg) subsets at peak of disease. We find that PD-1 restrains the generation and suppressive function of these effector Treg. This tissue eTreg signature is also observed in human and mouse Treg subsets from the tumor microenvironment, and are potent suppressor populations. Further studies are underway to determine how PD-1 impacts TCR signaling and FoxP3 transcriptional co-activators in Treg cells. Our study provides deeper insights into how PD-1 regulates T cell tolerance, underscoring the important role of PD-1 in regulating Treg differentiation and suppressive function during autoimmune diseases, and provide mechanistic insights for PD-1 modulation in cancer and autoimmunity. Supported by NIH P01AI108545

Protein Kinase Inhibitor-Mediated Immunoprophylactic and Immunotherapeutic Control of Colon Cancer.

Immunotherapy has allowed major advances in oncology in the past years, in particular with the development of immune checkpoint inhibitors, but the clinical benefits are still limited, particularly in colorectal cancer (CRC). Our scientific approach is based on the search for innovative immunotherapy with a final goal that aims to induce an effective antitumor immune response in CRC. Here, we focused on a multikinase inhibitor, H89. We carried out in vivo experiments based on syngeneic mouse models of colon cancer in BALB/c mice and chemically colon tumorigenesis. Flow cytometry, RNAseq, RT-qPCR, antibody-specific immune cell depletion, and Western blot were used to identify the immune cell type involved in the preventive and antitumor activity of H89. We demonstrated that H89 delays colon oncogenesis and prevents tumor growth. This latter effect seems to involve NK cells. H89 also inhibits colon tumor growth in a T-cell-dependent manner. Analysis of the immune landscape in the tumor microenvironment showed an increase of CD4+ Th1 cells and CD8+ cytotoxic T cells but a decrease of CD4+ Treg cell infiltration. Mechanistically, we showed that H89 could promote naïve CD4+ T-cell differentiation into Th1, a decrease in Treg differentiation, and an increase in CD8+ T-cell activation and cytotoxicity ex vivo. Furthermore, H89 induced overexpression of genes involved in antitumor immune response, such as IL-15RA, which depletion counteracts the antitumor effect of H89. We also found that H89 regulated Akt/PP2A pathway axis, involved in TCR and IL-15 signaling transduction. Our findings identify the H89 as a potential strategy for immune system activation leading to the prevention and treatment of CRC.

Your Regulatory T Cells Are What You Eat: How Diet and Gut Microbiota Affect Regulatory T Cell Development

Modern industrial practices have transformed the human diet over the last century, increasing the consumption of processed foods. Dietary imbalance of macro- and micro-nutrients and excessive caloric intake represent significant risk factors for various inflammatory disorders. Increased ingestion of food additives, residual contaminants from agricultural practices, food processing, and packaging can also contribute deleteriously to disease development. One common hallmark of inflammatory disorders, such as autoimmunity and allergies, is the defect in anti-inflammatory regulatory T cell (Treg) development and/or function. Treg represent a highly heterogeneous population of immunosuppressive immune cells contributing to peripheral tolerance. Tregs either develop in the thymus from autoreactive thymocytes, or in the periphery, from naïve CD4+ T cells, in response to environmental antigens and cues. Accumulating evidence demonstrates that various dietary factors can directly regulate Treg development. These dietary factors can also indirectly modulate Treg differentiation by altering the gut microbiota composition and thus the production of bacterial metabolites. This review provides an overview of Treg ontogeny, both thymic and peripherally differentiated, and highlights how diet and gut microbiota can regulate Treg development and function.

2′, 4′-Dihydroxy-2,3-dimethoxychalcone: A pharmacological inverse agonist of RORγt ameliorating Th17-driven inflammatory diseases by regulating Th17/Treg

Multiple sclerosis, inflammatory bowel disease and organ transplant rejection are related to Th17 cell development and inflammatory respond. RORγt, a specific transcription factor regulating Th17 cell differentiation, is a pivotal target for the treatment of diseases. However, the clinical application of RORγt inverse agonists reported so far has been hindered due to limited efficacy and toxic side effects. Plant-derived natural products with drug-like properties and safety are wide and valuable resources for candidate drug discovery. Herein, structure-based virtual screening was used to find out 2′,4′-Dihydroxy-2,3-dimethoxychalcone (DDC), a chalcone derivative rich in plants and food, located in the binding pocket of RORγt and targeted to inhibit RORγt activity. DDC repressed murine Th17 differentiation and promoted Treg differentiation remarkably in a dose-dependent manner. In addition, DDC treatment improved experimental autoimmune encephalomyelitis recovery, ameliorated experimental colitis severity, and prevented graft rejection significantly. Mechanically, DDC indirectly stabilized Foxp3 expression by inhibiting RORγt activity and the expression of its target gene profile in vitro and in vivo, which realized its regulation of Th17/Treg balance. In conclusion, our study provides a scientific basis that DDC, as an inverse agonist of RORγt with simple structure, rich sources, low cost, high efficiency, and low toxicity, has great potential for the development of a novel effective immunomodulator for the treatment of Th17-mediated inflammatory diseases.

Tumor-derived exosomes drive pre-metastatic niche formation in lung via modulating CCL1+ fibroblast and CCR8+ Treg cell interactions.

Since the lung is one of the most common sites for cancer metastasis, it could provide a suitable microenvironment for pre-metastatic niche (PMN) formation to facilitate tumor cell colonization. Regulatory T cells (Tregs) are an immunosuppressive cell type found ubiquitously in tumors and may play a crucial role in PNM formation. In this study, we investigated tumor-derived exosome (TDE)-induced Treg differentiation in the lung PMN as well as the underlying mechanisms. TDEs were isolated from the Lewis lung carcinoma cell line (LLC-exo) and their effects on mouse pulmonary fibroblasts was investigated in vitro as well as on lung tumor formation and metastasis in a pre-injected mouse model. Immune cell populations in the lung were analyzed by flow cytometry. Expression of CCL1 and CCR8 was evaluated by immunofluorescence staining, qRT-PCR and Western blot analyses. Cytokine expression was measured using mouse cytokine arrays and ELISA. The number of CD4+ FoxP3+ Tregs was significantly increased in lungs in a LLC-exo pre-injected mouse model. Lung fibroblasts secreted increased amounts of CCL1 after co-culture with LLC-exo, which induced Treg differentiation by activating its specific receptor CCR8, ultimately contributing to the establishment of an immunologically tolerant PMN. Moreover, inhibiting the release of LLC-exo by GW4869, or blocking the CCL1-CCR8 axis using AZ084, suppressed Tregs differentiation and tumor metastasis in the lung. Collectively, our study provides a novel mechanism by which Tregs are activated to form an immunologically tolerant PMN and demonstrates a critical link among lung fibroblasts, Tregs and metastatic tumor cells.

A novel strategy to investigate the factors regulating the Treg to Tfr transition during acute viral infection

Follicular regulatory T cells (Tfrs), a specialized subset of regulatory T cells (Tregs), have a particular role in the control of follicular helper T cell-driven germinal center (GC) responses. Following differentiation signals similar to those received by follicular helper T cells (Tfhs), Tfrs gain expression of characteristic chemokine receptors and transcription factors, such as CXCR5 and Bcl-6, allowing them to migrate into the B-cell follicle and perform in situ suppression. Thus, together with Tfhs, Tfrs help maintaining an optimized GC-reaction. However, the mechanism underlying the Treg-to-Tfr transition remains obscure. Here, we established a highly reproducible protocol for investigating the differentiation of Tregs into Tfrs by constructing spleen-chimeric mice combined with retrovirus transduction. We demonstrated that using this strategy, over 4 folds of Tregs could differentiate into Tfrs in Bcl-6 overexpression group compared to control counterparts (Migr1), and Bcl-6 could efficiently promote Tfr differentiation during acute viral infection. Hence, this method provides us an easy access to investigate the factors that regulate the differentiation program that converts Tregs into Tfrs, which will enhance our understanding of the networks regulating GC-reaction and shed new light on the molecular basis of immune homeostasis.

Genetic Pathways Activated in Heart Allograft Quilty Lesions

<h3>Purpose</h3> Quilty Lesions (QL) are enigmatic polytypic inflammatory cellular aggregates seen in some heart transplant biopsies. The study was designed to investigate genetic pathways that might be activated in heart transplant biopsies with QL. <h3>Methods</h3> Nine heart allograft biopsies from one institution were included in the study. Tissue was stained for H/E and biopsies separated as having QL (5) vs. no QL (4). Total RNA was then extracted from FFPE. RNA extraction and gene analysis was performed using the Nanostring n-Counter Human Organ Transplant Immunology Panel (Seattle, WA). <h3>Results</h3> The heat map (Figure 1) shows a clear separation between biopsies with QL and biopsies without QL. Pathways that were upregulated in biopsies with QL included: NK-kappa Beta, cytokine signaling, chemokine signaling, cytotoxicity, type II interferon, Toll-like receptor, B-cell receptor, T-cell checkpoint, lymphocyte trafficking, oxidative stress, MAPK, Class II Ag presentation, complement system, Class I antigen presentation, TNF family, NLR family, cell cycle regulation, mTOR, inflammasomes, TGF-beta, epigenetics and transcription, T-reg differentiation, Th1 differentiation, Th2 differentiation, Th17 differentiation, cytosolic DNS sensing, innate immune system, adaptive immune system, hematopoiesis, T-cell receptor signaling, cell-ECM interaction. In contrast, the following pathways were downregulated in biopsies with QL: autophagy, angiogenesis, tissue homeostasis. Biopsies with QL showed a predominance of adaptive type Th2 response over Th1genes (3.88x vs. 3.15x). Individual regulatory genes (all p< 0.001) that were higher in biopsies with QL included TGFbeta1, IL10RA, Foxp3 and JAK3. <h3>Conclusion</h3> Heart allograft biopsies with QL showed a clearly differential expression of genes compared to biopsies without QL. Pathways and individual upregulated genes suggest a shift towards regulation and acceptance of the allograft. QL, far from being passive bystanders, may have a positive immunomodulatory function in cardiac allografts.

Intermittent Exposure of Hypercapnia Suppresses Allograft Rejection via Induction of Treg Differentiation and Inhibition of Neutrophil Accumulation.

Background: In the management of major burn wounds, allogeneic skin transplantation is a critical procedure to improve wound repair. Our previous works found that intermittent exposure to carbon dioxide leads to permissive hypercapnia (HCA) and prolongs skin allograft survival. However, the modulatory effects of HCA exposure on the immune system are not well understood. Objectives: Our purpose was to investigate how intermittent exposure to HCA can effectively reduce the immune reaction to allogeneic skin graft rejection. Methods: A fully major histocompatibility complex-incompatible skin transplant from BALB/c to C57BL/6 mice model was utilized. Immune cells from splenic and draining lymph nodes were analyzed by flow cytometry. Serum proinflammatory cytokines were analyzed by ELISA. Results: Serum levels of IFN-γ, IL-2, IL-6, and TNF-α were significantly decreased in the HCA group. Additionally, the percentage of CD8+ cells in draining lymph nodes was significantly lower in HCA than in the control group. Moreover, the generation rate of FoxP3+ regulatory T cells (Tregs) from spleen naïve CD4+ T cells was increased by intermittent exposure to carbon dioxide. The infiltrated neutrophils were also eliminated by HCA. Taken together, we concluded that intermittent hypercapnia exposure could effectively suppress skin rejection by stimulating Treg cell generation and suppressing immune reactions.

A peptide encoded by pri-miRNA-31 represses autoimmunity by promoting Treg differentiation.

Recent evidence has revealed that small polypeptides (containing fewer than 100 amino acids) can be translated from noncoding RNAs (ncRNAs), which are usually defined as RNA molecules that do not encode proteins. However, studies on functional products translated from primary transcripts of microRNA (pri-miRNA) are quite limited. Here, we describe a peptide termed miPEP31 that is encoded by pri-miRNA-31. miPEP31 is highly expressed in Foxp3+ regulatory T cells (Tregs ) and significantly promotes the differentiation of Tregs without affecting their inhibitory ability. Our results show that miPEP31 is a cell-penetrating peptide both in vitro and in vivo. miPEP31 downregulates miR-31 expression, enhances peripheral Treg induction, and dramatically suppresses experimental autoimmune encephalomyelitis. Mechanistically, we show that miPEP31 acts as a transcriptional repressor inhibiting the expression of miRNA-31, a negative regulator of Tregs . Our results reveal an indispensable role of miPEP31 in maintaining immune homeostasis by promoting Treg differentiation and also present a potential therapeutic peptide for modulating miRNA expression and treating autoimmune diseases.

Functions of animal microRNA-encoded peptides: the race is on!

Short open reading frame (sORF)-encoded peptides (SEPs) recently emerged as new key players in biology. Pioneering work first established that sORFs encoded by long non-coding RNAs (lncRNAs) are efficiently translated and produce functional peptides. In plants, primary transcripts of microRNAs (pri-miRNAs) also produce sORF-encoded peptides called miPEPs, which are involved in specific transcriptional autoregulatory feedback loops (Lauressergues etal, 2015). To what extend are such mechanisms conserved in other species, especially in animals? In this issue of EMBO reports, Zhou etal show that pri-miR-31 encodes a miPEP promoting Treg differentiation and downregulating pri-miR-31 expression (Zhou etal, 2022).

Transforming growth factor-β1 in regulatory T cell biology.

Transforming growth factor-β1 (TGF-β1) is inextricably linked to regulatory T cell (Treg) biology. However, precisely untangling the role for TGF-β1 in Treg differentiation and function is complicated by the pleiotropic and context-dependent activity of this cytokine and the multifaceted biology of Tregs. Among CD4+ T cells, Tregs are the major producers of latent TGF-β1 and are uniquely able to activate this cytokine via expression of cell surface docking receptor glycoprotein A repetitions predominant (GARP) and αv integrins. Although a preponderance of evidence indicates no essential roles for Treg-derived TGF-β1 in Treg immunosuppression, TGF-β1 signaling is crucial for Treg development in the thymus and periphery. Furthermore, active TGF-β1 instructs the differentiation of other T cell subsets, including TH17 cells. Here, we will review TGF-β1 signaling in Treg development and function and discuss knowledge gaps, future research, and the TGF-β1/Treg axis in the context of cancer immunotherapy and fibrosis.

Functional Diversities of Regulatory T Cells in the Context of Cancer Immunotherapy.

Regulatory T cells (Tregs) are a subset of CD4+ T cells with their immunosuppressive activities to block abnormal or excessive immune responses to self and non-autoantigens. Tregs express the transcription factor Foxp3, maintain the immune homeostasis, and prevent the initiation of anti-tumor immune effects in various ways as their mechanisms to modulate tumor development. Recognition of different phenotypes and functions of intratumoral Tregs has offered the possibilities to develop therapeutic strategies by selectively targeting Tregs in cancers with the aim of alleviating their immunosuppressive activities from anti-tumor immune responses. Several Treg-based immunotherapeutic approaches have emerged to target cytotoxic T lymphocyte antigen-4, glucocorticoid-induced tumor necrosis factor receptor, CD25, indoleamine-2, 3-dioxygenase-1, and cytokines. These immunotherapies have yielded encouraging outcomes from preclinical studies and early-phase clinical trials. Further, dual therapy or combined therapy has been approved to be better choices than single immunotherapy, radiotherapy, or chemotherapy. In this short review article, we discuss our current understanding of the immunologic characteristics of Tregs, including Treg differentiation, development, therapeutic efficacy, and future potential of Treg-related therapies among the general cancer therapy.

Immunopathogenesis of endometriosis: An overview of the role of innate and adaptive immune cells and their mediators.

Endometriosis is a chronic inflammatory disease associated with the growth and proliferation of endometrial-like tissues outside the uterus. Although the exact etiology and mechanism of the pathogenesis of the disease have not been fully elucidated, the immune system cells and the mediators produced by them can be named as effective factors in the onset and progression of the disease. We aim to attempt to review studies on the role of the immune system in endometriosis to better understand the pathogenesis of endometriosis. Abundant production of inflammatory mediators by neutrophils and macrophages and reduced cytotoxicity of defined cells promote endometriosis at the early stages of the disease. Following an increase in the inflammation of the environment, the body takes compensatory mechanisms to reduce inflammation and establish homeostasis. For this purpose, the body produces remodeling and anti-inflammatory factors leading to slow conversion of the inflammatory environment into a non-inflammatory environment with proliferative and immunosuppressive properties. Environmental conditions induce M2 macrophages, TH2 cells, and Tregs differentiation, promoting disease progression by producing angiogenic and immunosuppressive factors. However, the exact molecular mechanism involved in changing inflammatory to non-inflammatory conditions is not yet fully understood. Due to the common characteristics of endometriotic cells and cancer cells, most potential treatment options for endometriosis have been suggested due to the results of these methods in the treatment of cancer. In this pathway, immune system cells and soluble mediators can be used as targets.

Multi-material basis and multi-mechanisms of the Dahuang Zhechong pill for regulating Treg/Th1 balance in hepatocellular carcinoma

BackgroundDahuang Zhechong pill (DHZCP) improves the inhibitory immune status of mice with hepatocellular carcinoma (HCC) by regulating Treg/Th1 balance. Hypothesis/PurposeTo study the multi-material basis and multi-mechanisms of DHZCP against HCC by regulating Treg/Th1 balance in vitro and in vivo. MethodsUPLC-MS/MS was used to detect the dynamic changes in 29 characteristic components of different polar parts of DHZCP. H&E and TUNEL were used to check pathological condition in HCC mice. The number of CD4+T, CD8+T, Treg, Th1, and Th1-like Treg cells was counted by flow cytometry. TGF-β, IL-10, IFN-γ, and TNF-α content were detected by ELISA. α-Ketoglutarate and glutamine levels were detected by Trace1310/TSQ8000 GC-MS/MS. p-Smad2, and p-Smad3 protein levels were detected by WB, mRNA expression of Smad2, alanine-serine-cysteine transporter-2, glutaminase, and glutamate dehydrogenase were detected by RT-PCR. Simca-p multivariate data analysis software was used to evaluate the relationship between the different polar parts of DHZCP and the proportion of Treg cells. ResultsWater-soluble (PW) and ethyl acetate (PE) polar parts of DHZCP affected the HCC immune system by inhibiting the differentiation of Tregs, reversing the balance of Treg/Th1, and significantly reduced the tumor volume and weight. However, petroleum ether and n-butanol polar parts had no above actions. The changes in emodin, chrysophanol, aloe vera emodin, emodin-8-O-β-D-glycoside, gallic acid, naringenin, baicalein, wogonin, norwogonin, apigenin, chrysin, glycyrrhizin, formononetin, and palmitic acid were closely related to the changes of Treg cells, which is the main material basis of DHZCP inhibition of Treg differentiation. Additionally, PW mainly inhibit the differentiation of Treg cells by affecting the metabolism of hepatoma cells, improving tumor microenvironment acidity, and glutamine depletion. However, PE inhibited the differentiation of Treg cells mainly by regulating the TGF-β/Smad pathway. ConclusionIn this study, accurate analysis of multi-component was combined with pharmacodynamic evaluations to identify the pharmacodynamic substances of DHZCP in regulating Treg/Th1 balance, and clarified the multi-target mechanism of DHZCP to improve tumor immunity. The study style offers a novel approach for pharmacological research on TCM.

S1P analogues SEW2871, BAF312 and FTY720 affect human Th17 and Treg generation ex vivo

Multiple Sclerosis is an immune-mediated neurodegenerative disease. IL-23-mediated signaling and Th17 cells play critical roles in disease pathogenesis in murine models of disease and humans. Sphingosine 1 phosphate (S1P) regulates migration of several types of immune cells including Th17 cells. S1P analogues (fingolimod (FTY720) and Siponimod (BAF312)) have been approved and currently used for MS treatment. Immunomodulatory roles for FTY720 have been defined, however, how different S1P analogues impact human Th17 and Treg cell generation and cytokine production, and IL-23-mediated signaling have not yet been explored in detail. In the current study, we investigated the effects of S1P receptor 1 (S1P1) specific S1P analogue SEW2871, S1P1 and S1P5 specific BAF312, and non-selective FTY720 on human Th17 and Treg differentiation and IL-23-mediated signaling. All three S1P analogues directly inhibited Th17 cell differentiation ex vivo while increasing Treg differentiation from naive CD4 + T cells. All three S1P analogues suppressed IL-23-mediated STAT4, NF-kB and AKT activation. Lastly, all three S1P analogues also inhibited Dectin-1 expression by both mature and immature monocyte-derived dendritic cells (moDCs) and in turn curdlan-mediated production of IL-23p19, p40, IL-6 and IL-1β cytokines. Our results provide novel insight into the immunomodulatory roles of different S1P analogues on human Th17 and Treg cell biology.