Selective IL-27 production by intestinal regulatory T cells permits gut-specific regulation of TH17 cell immunity.

Regulatory T cells (Treg cells) are instrumental in establishing immunological tolerance. However, the precise effector mechanisms by which Treg cells control a specific type of immune response in a given tissue remains unresolved. By simultaneously studying Treg cells from different tissue origins under systemic autoimmunity, in the present study we show that interleukin (IL)-27 is specifically produced by intestinal Treg cells to regulate helper T17 cell (TH17 cell) immunity. Selectively increased intestinal TH17 cell responses in mice with Treg cell-specific IL-27 ablation led to exacerbated intestinal inflammation and colitis-associated cancer, but also helped protect against enteric bacterial infection. Furthermore, single-cell transcriptomic analysis has identified a CD83+CD62Llo Treg cell subset that is distinct from previously characterized intestinal Treg cell populations as the main IL-27 producers. Collectively, our study uncovers a new Treg cell suppression mechanism crucial for controlling a specific type of immune response in a particular tissue and provides further mechanistic insights into tissue-specific Treg cell-mediated immune regulation.

miR-15/16 clusters restrict effector Treg cell differentiation and function.

Effector regulatory T cells (eTregs) exhibit distinct homeostatic properties and superior suppressor capacities pivotal for controlling immune responses mediated by their conventional T cell counterpart. While the role of microRNAs (miRNAs) in Tregs has been well-established, how miRNAs regulate eTregs remains poorly understood. Here, we demonstrate that miR-15/16 clusters act as key regulators in limiting eTreg responses. Loss of miR-15/16 clusters leads to increased eTreg frequencies with enhanced suppressor function. Consequently, mice with Treg-specific ablation of miR-15/16 clusters display attenuated immune responses during neuroinflammation and upon both infectious and non-infectious challenges. Mechanistically, miR-15/16 clusters exert their regulatory effect in part through repressing IRF4, a transcription factor essential for eTreg differentiation and function. Moreover, miR-15/16 clusters also directly target neuritin, an IRF4-dependent molecule, known for its role in Treg-mediated regulation of plasma cell responses. Together, we identify an miRNA family that controls an important Treg subset and further demonstrate that eTreg responses are tightly regulated at both transcriptional and posttranscriptional levels.

A Distance Transformation Deep Forest Framework With Hybrid-Feature Fusion for CXR Image Classification.

Detecting pneumonia, especially coronavirus disease 2019 (COVID-19), from chest X-ray (CXR) images is one of the most effective ways for disease diagnosis and patient triage. The application of deep neural networks (DNNs) for CXR image classification is limited due to the small sample size of the well-curated data. To tackle this problem, this article proposes a distance transformation-based deep forest framework with hybrid-feature fusion (DTDF-HFF) for accurate CXR image classification. In our proposed method, hybrid features of CXR images are extracted in two ways: hand-crafted feature extraction and multigrained scanning. Different types of features are fed into different classifiers in the same layer of the deep forest (DF), and the prediction vector obtained at each layer is transformed to form distance vector based on a self-adaptive scheme. The distance vectors obtained by different classifiers are fused and concatenated with the original features, then input into the corresponding classifier at the next layer. The cascade grows until DTDF-HFF can no longer gain benefits from the new layer. We compare the proposed method with other methods on the public CXR datasets, and the experimental results show that the proposed method can achieve state-of-the art (SOTA) performance. The code will be made publicly available at https://github.com/hongqq/DTDF-HFF.

Selective IL-27 production by intestinal regulatory T cells permits gut-specific regulation of Th17 immunity.

Regulatory T (Treg) cells are instrumental in establishing immunological tolerance. However, the precise effector mechanisms by which Treg cells control a specific type of immune response in a given tissue remains unresolved. By simultaneously studying Treg cells from different tissue origins under systemic autoimmunity, here we show that IL-27 is specifically produced by intestinal Treg cells to regulate Th17 immunity. Selectively increased intestinal Th17 responses in mice with Treg cell-specific IL-27 ablation led to exacerbated intestinal inflammation and colitis-associated cancer, but also helped protect against enteric bacterial infection. Furthermore, single-cell transcriptomic analysis has identified a CD83+TCF1+ Treg cell subset that is distinct from previously characterized intestinal Treg cell populations as the main IL-27 producers. Collectively, our study uncovers a novel Treg cell suppression mechanism crucial for controlling a specific type of immune response in a particular tissue, and provides further mechanistic insights into tissue-specific Treg cell-mediated immune regulation.

miR-494-5p mediates the antioxidant activity of EPA by targeting the mitochondrial elongation factor 1 gene MIEF1 in HepG2 cells.

Eicosapentaenoic acid (EPA) shows antioxidant activity, which may be attributed to its regulatory effect on microRNA expression. Our preliminary study indicated that EPA upregulated miR-494-5p, which was possibly involved in the regulation of cellular stress responses. The current study aimed to address whether miR-494-5p was targeted by EPA to regulate cellular oxidative stress and its possible functional mechanism. The results showed that miR-494-5p mediated the antioxidant effect of EPA and miR-494-5p reduction deteriorated EPA-induced increase in the cellular antioxidant capacity of HepG2 cells. Moreover, the mitochondrial elongation factor 1 (MIEF1) gene was a target gene of miR-494-5p. Both miR-494-5p overexpression and MIEF1 knockdown significantly enhanced cellular antioxidant capacity, as indicated by a reduction in the reactive oxygen species level and an increase in the total cellular antioxidant capacity, along with enhancing antioxidant enzymes. Thus, miR-494-5p and MIEF1 had opposite effects on cellular antioxidant capacity. Furthermore, their regulatory effects on oxidative stress may have been attributed to modulation of mitochondrial function, biogenesis and homeostasis. Taken together, the findings indicated that miR-494-5p mediated EPA activity and promoted cellular antioxidant capacity by inhibiting the expression of MIEF1, which further modulated mitochondrial structure and activity. This study may provide novel insights into the post-translational regulation of antioxidation reactions, which involves the coordinated control of mitochondria.

Evaluation of retinal vascular density and related factors using OCTA in children and adolescents with myopia without maculopathy.

BACKGROUND: Myopia is the most common ophthalmic condition worldwide with a rapidly increasing prevalence. This study aimed to compare the retinal microvasculature in the superficial capillary plexus (SCP) in children and adolescents with mild and moderate/high myopia using optical coherence tomography angiography, determine the relationship between retinal parameters and axial length (AL), and understand the occurrence and progression of myopia in microcirculation. METHODS: This prospective observational study included 39 participants with mild myopia and 33 participants with moderate/high myopia. Vessel density (VD) and perfusion density (PD) in the SCP, the foveal avascular zone (FAZ), and AL were compared between the groups and the relationship between these retinal parameters and AL was assessed. RESULTS: No difference in SCP VD or PD was observed between the two groups. The FAZ did not differ significantly between groups whereas significant differences in age, height, refractive status, and AL were observed. Significantly shorter AL was observed in participants with mild myopia compared with the moderate/high myopia group. Age was positively correlated with height (r = 0.852) and refractive status was negatively correlated with AL (r = -0.588). AL was positively correlated with VD (r = 0.317) and PD (r = 0.308) in the SCP and AL was negatively correlated with the FAZ (r = -0.434). CONCLUSIONS: This study revealed that superficial foveal microvessel density was unaffected in children and adolescents without pathological myopia. However, myopia progression was associated with a change in AL, and an AL increase altered macular blood flow.

MiR-125b-5p is targeted by curcumin to regulate the cellular antioxidant capacity.

As a natural polyphenolic food supplement and the principal curcuminoid in turmeric, curcumin shows antioxidant, anti-inflammatory, and antitumor activities. However, its specific functional mechanism remains unclear. Our preliminary study indicated that miR-125b-5p was downregulated by a curcumin extract. This study aimed to determine whether miR-125b-5p is involved in the antioxidant regulation of curcumin. The results showed that miR-125b-5p overexpression had a pro-oxidant effect by reducing the cellular antioxidant capacity, as well as decreasing the activities of catalase (CAT) and superoxide dismutase (SOD) in the normal liver cell line LO2. However, miR-125b-5p repression significantly increased the cellular antioxidant capacity and enhanced the activities of CAT and SOD. Further investigation demonstrated that the cellular antioxidant capacity induced by curcumin extract was inhibited by miR-125b-5p overexpression. Thus, curcumin may exhibit antioxidant effects by repressing miR-125b-5p expression, which provides new insights into the molecular antioxidant mechanism of curcumin and other functional food components.

Analysis of macular blood flow changes in thyroid associated ophthalmopathy.

BACKGROUND: To evaluate the changes in macular superficial retinal vessel density and their relation with visual acuity in thyroid-associated ophthalmopathy (TAO) patients with different severity. METHODS: This cross-sectional observational study included 70 TAO patients and 70 healthy controls. Only data from the right eyes were analyzed. TAO patients were divided into 7 subgroups according to the NOSPECS score. Foveal avascular zone (FAZ), vascular density (VD), and perfusion density (PD) of macular 1 mm diameter and 6 mm diameter areas were measured by optical coherence tomography angiography (OCTA). RESULTS: In TAO patients, significant increases were found in macular and foveal vascular densities (FVD) and perfusion densities (FPD) while a significant decrease was found in the FAZ area when compared with the control group (p < 0.05). Spearman correlation analysis and multiple linear regression analysis showed that TAO severity grade was negatively correlated with FVD (ß = -1.150, p = 0.032), FPD (ß = -0.024, p = 0.042), MVD (ß = -0.583, p = 0.020) and MPD (ß = -0.011, p = 0.010). Clinical activity score (CAS) score showed positive correlation with FVD (ß = 0.794, p = 0.035) and FPD(ß = 0.017, p = 0.041). FVD (ß = -0.009, p = 0.033), MVD(ß = -0.034, p < 0.001), FPD(ß = -0.416, p = 0.039) and MPD(ß = -2.428, p < 0.001) all showed negative correlation with best corrected visual acuity (BCVA). CONCLUSIONS: There was an overall increase in superficial macular blood flow in TAO patients compared with healthy controls and the blood flow decreased as TAO got worse. Superficial macular flow density was negatively correlated with BCVA.

MRDFF: A deep forest based framework for CT whole heart segmentation.

Automatic whole heart segmentation plays an important role in the treatment and research of cardiovascular diseases. In this paper, we propose an improved Deep Forest framework, named Multi-Resolution Deep Forest Framework (MRDFF), which accomplishes whole heart segmentation in two stages. We extract the heart region by binary classification in the first stage, thus avoiding the class imbalance problem caused by too much background. The results of the first stage are then subdivided in the second stage to obtain accurate cardiac substructures. In addition, we also propose hybrid feature fusion, multi-resolution fusion and multi-scale fusion to further improve the segmentation accuracy. Experiments on the public dataset MM-WHS show that our model can achieve comparable accuracy in about half the training time of neural network models.

Gut epithelial IL-27 confers intestinal immunity through the induction of intraepithelial lymphocytes.

IL-27 controls a diverse range of immune responses in many disease settings. Here, we identify intestinal epithelial cells (IECs) as one of the major IL-27 cellular sources in the gut-associated tissue. Unlike IL-27 secreted by innate immune cells, gut epithelial IL-27 is dispensable for T-bet+ regulatory T cell (T reg cell) differentiation or IL-10 induction. Rather, IEC-derived IL-27 specifically promotes a distinct CD8αα+CD4+ intraepithelial lymphocyte (IEL) population that acquires their functional differentiation at the intestinal epithelium. Loss of IL-27 in IECs leads to a selective defect in CD8αα+CD4+ IELs over time. Consequently, mice with IEC-specific IL-27 ablation exhibited elevated pathogen burden during parasitic infection, and this could be rescued by transfer of exogenous CD8αα+CD4+ IELs. Collectively, our data reveal that in addition to its known regulatory properties in preventing immune hyperactivity, gut epithelial IL-27 confers barrier immunity by inducing a specific IEL subset and further suggest that IL-27 produced by different cell types plays distinct roles in maintaining intestinal homeostasis.

A direct slicing technique for the 3D printing of implicitly represented medical models.

In conventional medical image printing methods, volumetric medical data needs to be conversed into STereo Lithography (STL) format, the most commonly used format for representing geometric models for 3D printing. However, this STL conversion process is not only time consuming, but more importantly, it often leads to the loss of accuracy. It has become a critical factor hindering the printing efficiency and precision of organ models. By examining the key characteristics of discrete medical volume data, this paper proposes a direct slicing technique for printing implicitly represented 3D medical models. The proposed method mainly consists of three algorithms: (1) A layer-based contour extraction algorithm for discrete volume data; (2) An inner shell construction algorithm based on discrete point differential indentation; (3) An infill generation algorithm based on the constructed virtual contour and scan lines. The proposed method has been applied to the slicing of several organ models for experiments, and the ratios of time cost and memory cost between the conventional method and the proposed method are about 4-100 and 1.1 to 1.4 respectively, which demonstrate that the proposed method has a great improvement in both time and space performance when compared with the conventional STL-based method. Our technique extends the direct input format of geometric models for additive manufacturing. That is, discrete volume data can be used as a direct input for additive manufacturing without conversion to STL format.

miR-155 promotes T reg cell development by safeguarding medullary thymic epithelial cell maturation.

During thymocyte development, medullary thymic epithelial cells (mTECs) provide appropriate instructive cues in the thymic microenvironment for not only negative selection but also the generation of regulatory T (T reg) cells. Here, we identify that miR-155, a microRNA whose expression in T reg cells has previously been shown to be crucial for their development and homeostasis, also contributes to thymic T reg (tT reg) cell differentiation by promoting mTEC maturation. Mechanistically, we show that RANKL stimulation induces expression of miR-155 to safeguard the thymic medulla through targeting multiple known and previously uncharacterized molecules within the TGFß signaling pathway, which is recognized for its role in restricting the maturation and expansion of mTECs. Our work uncovers a miR-155-TGFß axis in the thymic medulla to determine mTEC maturity and, consequently, the quantity of tT reg cells and suggests that miR-155 ensures proper tT reg cell development in both cell-intrinsic and -extrinsic manners.

MiR-23~27~24-mediated control of humoral immunity reveals a TOX-driven regulatory circuit in follicular helper T cell differentiation.

Follicular helper T (TFH) cells are essential for generating protective humoral immunity. To date, microRNAs (miRNAs) have emerged as important players in regulating TFH cell biology. Here, we show that loss of miR-23~27~24 clusters in T cells resulted in elevated TFH cell frequencies upon different immune challenges, whereas overexpression of this miRNA family led to reduced TFH cell responses. Mechanistically, miR-23~27~24 clusters coordinately control TFH cells through targeting a network of genes that are crucial for TFH cell biology. Among them, thymocyte selection-associated HMG-box protein (TOX) was identified as a central transcription regulator in TFH cell development. TOX is highly up-regulated in both mouse and human TFH cells in a BCL6-dependent manner. In turn, TOX promotes the expression of multiple molecules that play critical roles in TFH cell differentiation and function. Collectively, our results establish a key miRNA regulon that maintains optimal TFH cell responses for resultant humoral immunity.

Differential cell-intrinsic regulations of germinal center B and T cells by miR-146a and miR-146b.

Reciprocal interactions between B and follicular T helper (Tfh) cells orchestrate the germinal center (GC) reaction, a hallmark of humoral immunity. Abnormal GC responses could lead to the production of pathogenic autoantibodies and the development of autoimmunity. Here we show that miR-146a controls GC responses by targeting multiple CD40 signaling pathway components in B cells; by contrast, loss of miR-146a in T cells does not alter humoral responses. However, specific deletion of both miR-146a and its paralog, miR-146b, in T cells increases Tfh cell numbers and enhanced GC reactions. Thus, our data reveal differential cell-intrinsic regulations of GC B and Tfh cells by miR-146a and miR-146b. Together, members of the miR-146 family serve as crucial molecular brakes to coordinately control GC reactions to generate protective humoral responses without eliciting unwanted autoimmunity.

A Novel miR-24-TCF1 Axis in Modulating Effector T Cell Responses.

miR-23∼27∼24 was recently implicated in restricting Th2 immunity, as well as the differentiation and function of other effector T cell lineages. Interestingly, miR-24, unlike other family members, actually promotes Th1 and Th17 responses. In this article, we show that miR-24 drives the production of IFN-γ and IL-17 in T cells at least in part through targeting TCF1, a transcription factor known for its role in limiting Th1 and Th17 immunity. Surprisingly, whereas TCF1 was previously shown to promote Th2 responses through inducing GATA3, enforced TCF1 expression in miR-24-overexpressing T cells led to further downregulation of IL-4 and GATA3 expression, suggesting miR-24-mediated inhibition of Th2 immunity cannot be attributed to TCF1 repression by miR-24. Together, our data demonstrate a novel miR-24-TCF1 pathway in controlling effector cytokine production by T cells and further suggest miR-24 could function as a key upstream molecule regulating TCF1-mediated immune responses.

Excessive expression of miR-27 impairs Treg-mediated immunological tolerance.

MicroRNAs (miRs) are tightly regulated in the immune system, and aberrant expression of miRs often results in hematopoietic malignancies and autoimmune diseases. Previously, it was suggested that elevated levels of miR-27 in T cells isolated from patients with multiple sclerosis facilitate disease progression by inhibiting Th2 immunity and promoting pathogenic Th1 responses. Here we have demonstrated that, although mice with T cell-specific overexpression of miR-27 harbor dysregulated Th1 responses and develop autoimmune pathology, these disease phenotypes are not driven by miR-27 in effector T cells in a cell-autonomous manner. Rather, dysregulation of Th1 responses and autoimmunity resulted from a perturbed Treg compartment. Excessive miR-27 expression in murine T cells severely impaired Treg differentiation. Moreover, Tregs with exaggerated miR-27-mediated gene regulation exhibited diminished homeostasis and suppressor function in vivo. Mechanistically, we determined that miR-27 represses several known as well as previously uncharacterized targets that play critical roles in controlling multiple aspects of Treg biology. Collectively, our data show that miR-27 functions as a key regulator in Treg development and function and suggest that proper regulation of miR-27 is pivotal to safeguarding Treg-mediated immunological tolerance.

miR-23∼27∼24 clusters control effector T cell differentiation and function.

Coordinated repression of gene expression by evolutionarily conserved microRNA (miRNA) clusters and paralogs ensures that miRNAs efficiently exert their biological impact. Combining both loss- and gain-of-function genetic approaches, we show that the miR-23∼27∼24 clusters regulate multiple aspects of T cell biology, particularly helper T (Th) 2 immunity. Low expression of this miRNA family confers proper effector T cell function at both physiological and pathological settings. Further studies in T cells with exaggerated regulation by individual members of the miR-23∼27∼24 clusters revealed that miR-24 and miR-27 collaboratively limit Th2 responses through targeting IL-4 and GATA3 in both direct and indirect manners. Intriguingly, although overexpression of the entire miR-23 cluster also negatively impacts other Th lineages, enforced expression of miR-24, in contrast to miR-23 and miR-27, actually promotes the differentiation of Th1, Th17, and induced regulatory T cells, implying that under certain conditions, miRNA families can fine tune the biological effects of their regulation by having individual members antagonize rather than cooperate with each other. Together, our results identify a miRNA family with important immunological roles and suggest that tight regulation of miR-23∼27∼24 clusters in T cells is required to maintain optimal effector function and to prevent aberrant immune responses.

A Single miRNA-mRNA Interaction Affects the Immune Response in a Context- and Cell-Type-Specific Manner.

MicroRNA (miRNA)-dependent regulation of gene expression confers robustness to cellular phenotypes and controls responses to extracellular stimuli. Although a single miRNA can regulate expression of hundreds of target genes, it is unclear whether any of its distinct biological functions can be due to the regulation of a single target. To explore in vivo the function of a single miRNA-mRNA interaction, we mutated the 3' UTR of a major miR-155 target (SOCS1) to specifically disrupt its regulation by miR-155. We found that under physiologic conditions and during autoimmune inflammation or viral infection, some immunological functions of miR-155 were fully or largely attributable to the regulation of SOCS1, whereas others could be accounted only partially or not at all by this interaction. Our data suggest that the role of a single miRNA-mRNA interaction is dependent on cell type and biological context.

IFNγ signaling endows DCs with the capacity to control type I inflammation during parasitic infection through promoting T-bet+ regulatory T cells.

IFNγ signaling drives dendritic cells (DCs) to promote type I T cell (Th1) immunity. Here, we show that activation of DCs by IFNγ is equally crucial for the differentiation of a population of T-bet+ regulatory T (Treg) cells specialized to inhibit Th1 immune responses. Conditional deletion of IFNγ receptor in DCs but not in Treg cells resulted in a severe defect in this specific Treg cell subset, leading to exacerbated immune pathology during parasitic infections. Mechanistically, IFNγ-unresponsive DCs failed to produce sufficient amount of IL-27, a cytokine required for optimal T-bet induction in Treg cells. Thus, IFNγ signalling endows DCs with the ability to efficiently control a specific type of T cell immunity through promoting a corresponding Treg cell population.

Function of miR-146a in controlling Treg cell-mediated regulation of Th1 responses.

Foxp3(+) regulatory T (Treg) cells maintain immune homeostasis by limiting different types of inflammatory responses. Here, we report that miR-146a, one of the miRNAs prevalently expressed in Treg cells, is critical for their suppressor function. The deficiency of miR-146a in Treg cells resulted in a breakdown of immunological tolerance manifested in fatal IFNγ-dependent immune-mediated lesions in a variety of organs. This was likely due to augmented expression and activation of signal transducer and activator transcription 1 (Stat1), a direct target of miR-146a. Likewise, heightened Stat1 activation in Treg cells subjected to a selective ablation of SOCS1, a key negative regulator of Stat1 phosphorylation downstream of the IFNγ receptor, was associated with analogous Th1-mediated pathology. Our results suggest that specific aspects of Treg suppressor function are controlled by a single miRNA and that an optimal range of Stat1 activation is important for Treg-mediated control of Th1 responses and associated autoimmunity.