Antitumor progenitor exhausted CD8+ T cells are sustained by TCR engagement.

The durability of an antitumor immune response is mediated in part by the persistence of progenitor exhausted CD8+ T cells (Tpex). Tpex serve as a resource for replenishing effector T cells and preserve their quantity through self-renewal. However, it is unknown how T cell receptor (TCR) engagement affects the self-renewal capacity of Tpex in settings of continued antigen exposure. Here we use a Lewis lung carcinoma model that elicits either optimal or attenuated TCR signaling in CD8+ T cells to show that formation of Tpex in tumor-draining lymph nodes and their intratumoral persistence is dependent on optimal TCR engagement. Notably, attenuated TCR stimulation accelerates the terminal differentiation of optimally primed Tpex. This TCR-reinforced Tpex development and self-renewal is coupled to proximal positioning to dendritic cells and epigenetic imprinting involving increased chromatin accessibility at Egr2 and Tcf1 target loci. Collectively, this study highlights the critical function of TCR engagement in sustaining Tpex during tumor progression.

Regulatory T cells function in established systemic inflammation and reverse fatal autoimmunity.

The immunosuppressive function of regulatory T (Treg) cells is dependent on continuous expression of the transcription factor Foxp3. Foxp3 loss of function or induced ablation of Treg cells results in a fatal autoimmune disease featuring all known types of inflammatory responses with every manifestation stemming from Treg cell paucity, highlighting a vital function of Treg cells in preventing fatal autoimmune inflammation. However, a major question remains whether Treg cells can persist and effectively exert their function in a disease state, where a broad spectrum of inflammatory mediators can either inactivate Treg cells or render innate and adaptive pro-inflammatory effector cells insensitive to suppression. By reinstating Foxp3 protein expression and suppressor function in cells expressing a reversible Foxp3 null allele in severely diseased mice, we found that the resulting single pool of rescued Treg cells normalized immune activation, quelled severe tissue inflammation, reversed fatal autoimmune disease and provided long-term protection against them. Thus, Treg cells are functional in settings of established broad-spectrum systemic inflammation and are capable of affording sustained reset of immune homeostasis.

A distal Foxp3 enhancer enables interleukin-2 dependent thymic Treg cell lineage commitment for robust immune tolerance.

Activation of the STAT5 transcription factor downstream of the Interleukin-2 receptor (IL-2R) induces expression of Foxp3, a critical step in the differentiation of regulatory T (Treg) cells. Due to the pleiotropic effects of IL-2R signaling, it is unclear how STAT5 acts directly on the Foxp3 locus to promote its expression. Here, we report that IL-2 - STAT5 signaling converged on an enhancer (CNS0) during Foxp3 induction. CNS0 facilitated the IL-2 dependent CD25+Foxp3- precursor to Treg cell transition in the thymus. Its deficiency resulted in impaired Treg cell generation in neonates, which was partially mitigated with age. While the thymic Treg cell paucity caused by CNS0 deficiency did not result in autoimmunity on its own, it exacerbated autoimmune manifestations caused by disruption of the Aire gene. Thus, CNS0 enhancer activity ensures robust Treg cell differentiation early in postnatal life and cooperatively with other tolerance mechanisms minimizes autoimmunity.

Control of the inheritance of regulatory T cell identity by a cis element in the Foxp3 locus.

In multicellular organisms, specialized functions are delegated to distinct cell types whose identity and functional integrity are maintained upon challenge. However, little is known about the mechanisms enabling lineage inheritance and their biological implications. Regulatory T (Treg) cells, which express the transcription factor Foxp3, suppress fatal autoimmunity throughout the lifespan of animals. Here, we show that a dedicated Foxp3 intronic element CNS2 maintains Treg cell lineage identity by acting as a sensor of the essential Treg cell growth factor IL-2 and its downstream target STAT5. CNS2 sustains Foxp3 expression during division of mature Treg cells when IL-2 is limiting and counteracts proinflammatory cytokine signaling that leads to the loss of Foxp3. CNS2-mediated stable inheritance of Foxp3 expression is critical for adequate suppression of diverse types of chronic inflammation by Treg cells and prevents their differentiation into inflammatory effector cells. The described mechanism may represent a general principle of the inheritance of differentiated cell states.

An essential role for the IL-2 receptor in Treg cell function.

Regulatory T cells (Treg cells), which have abundant expression of the interleukin 2 receptor (IL-2R), are reliant on IL-2 produced by activated T cells. This feature indicates a key role for a simple network based on the consumption of IL-2 by Treg cells in their suppressor function. However, congenital deficiency in IL-2R results in reduced expression of the Treg cell lineage-specification factor Foxp3, which has confounded experimental efforts to understand the role of IL-2R expression and signaling in the suppressor function of Treg cells. Using genetic gain- and loss-of-function approaches, we found that capture of IL-2 was dispensable for the control of CD4+ T cells but was important for limiting the activation of CD8+ T cells, and that IL-2R-dependent activation of the transcription factor STAT5 had an essential role in the suppressor function of Treg cells separable from signaling via the T cell antigen receptor.

Hippo Kinases Mst1 and Mst2 Sense and Amplify IL-2R-STAT5 Signaling in Regulatory T Cells to Establish Stable Regulatory Activity.

Interleukin-2 (IL-2) and downstream transcription factor STAT5 are important for maintaining regulatory T (Treg) cell homeostasis and function. Treg cells can respond to low IL-2 levels, but the mechanisms of STAT5 activation during partial IL-2 deficiency remain uncertain. We identified the serine-threonine kinase Mst1 as a signal-dependent amplifier of IL-2-STAT5 activity in Treg cells. High Mst1 and Mst2 (Mst1-Mst2) activity in Treg cells was crucial to prevent tumor resistance and autoimmunity. Mechanistically, Mst1-Mst2 sensed IL-2 signals to promote the STAT5 activation necessary for Treg cell homeostasis and lineage stability and to maintain the highly suppressive phosphorylated-STAT5+ Treg cell subpopulation. Unbiased quantitative proteomics revealed association of Mst1 with the cytoskeletal DOCK8-LRCHs module. Mst1 deficiency limited Treg cell migration and access to IL-2 and activity of the small GTPase Rac, which mediated downstream STAT5 activation. Collectively, IL-2-STAT5 signaling depends upon Mst1-Mst2 functions to maintain a stable Treg cell pool and immune tolerance.

Inflammation-induced repression of chromatin bound by the transcription factor Foxp3 in regulatory T cells.

The transcription factor Foxp3 is indispensable for the ability of regulatory T cells (Treg cells) to suppress fatal inflammation. Here we characterized the role of Foxp3 in chromatin remodeling and the regulation of gene expression in actively suppressive Treg cells in an inflammatory setting. Although genome-wide occupancy of regulatory elements in DNA by Foxp3 was similar in resting Treg cells and those activated in vivo, Foxp3-bound enhancer elements in the DNA were poised for repression only in activated Treg cells. Following activation, Foxp3-bound sites showed diminished accessibility of chromatin and selective deposition of histone H3 trimethylated at Lys27 (H3K27me3), which was associated with recruitment of the histone methyltransferase Ezh2 and downregulation of the expression of nearby genes. Thus, Foxp3 poises its targets for repression by facilitating the formation of repressive chromatin in Treg cells upon their activation in response to inflammatory cues.

Characterization of the MT-2 Treg-like cell line in the presence and absence of forkhead box P3 (FOXP3).

CD4+ forkhead box P3 (FOXP3)+ regulatory T cells (Tregs) are essential in maintaining immune tolerance and suppressing excessive immune responses. Tregs also contribute to tissue repair processes distinct from their roles in immune suppression. For these reasons, Tregs are candidates for targeted therapies for inflammatory and autoimmune diseases, and in diseases where tissue damage occurs. MT-2 cells, an immortalized Treg-like cell line, offer a model to study Treg biology and their therapeutic potential. In the present study, we use clustered regularly interspaced palindromic repeats (CRISPR)-mediated knockdown of FOXP3 in MT-2 cells to understand the transcriptional and functional changes that occur when FOXP3 is lost and to compare MT-2 cells with primary human Tregs. We demonstrate that loss of FOXP3 affects the transcriptome of MT-2 cells and that FOXP3's potential downstream targets include a wide range of transcripts that participate in the cell cycle, promote growth and contribute to inflammatory processes, but do not wholly simulate previously reported human primary Treg transcriptional changes in the absence of FOXP3. We also demonstrate that FOXP3 regulates cell cycling and proliferation, expression of molecules crucial to Treg function and MT-2 cell-suppressive activities. Thus, MT-2 cells offer opportunities to address regulatory T-cell functions in vitro.

Evaluation of the safety and efficacy of a fractional picosecond 1064 nm laser for post-acne erythema in adult Chinese patients.

OBJECTIVE: To evaluate the efficacy and safety of fractional 1064 nm picosecond Nd:YAG laser (FPNYL) in the treatment of post-acne erythema (PAE) of adult Chinese. MATERIALS AND METHODS: A total of 22 patients received 1 session of treatment and were followed up at the eighth week. Primary outcomes were measured by the Clinician erythema assessment scale (CEAS). Secondary outcomes included a global aesthetic improvement scale (GAIS) and patients' assessment of satisfaction on a five-point scale. Pain scores and adverse effects were also evaluated. RESULTS: Twenty-two patients with Fitzpatrick skin types III and IV were enrolled in the study and completed all treatments and follow-up visits. The mean CEAS scores fell from 2.74 ± 0.80 to 1.95 ± 0.75 (p < 0.05). The mean GAIS of PAE improvement was 2.46 ± 0.68. Erythema percentile scores by VISIA increased from 32.63 ± 7.0 to 45.75 ± 11.45 (t = 5.442, p = 0). The patient satisfaction score was 1.86 ± 1.17. The pain scores were 3.27 ± 1.17 for the FPNYL treatment (varied from 2 to 6). There were moderate erythema and oedema, which last for 3.84 ± 0.78 days. There were overall 68.18% (15/22) patients who felt pruritus in different degrees and 27.27% patients who encountered acne eruptions (white head type). No scar, hyperpigmentation or hypopigmentation was found. CONCLUSION: Treatment with fractional 1064 nm picosecond Nd:YAG laser is effective and safe for PAE of Chinese patients.

Controlled Synthesis of V-Doped Heterogeneous Ni3S2/NiS Nanorod Arrays as Efficient Hydrogen Evolution Electrocatalysts.

The development of low-cost and high-efficient electrocatalysts toward hydrogen evolution reaction (HER) is highly desirable and challenging. Herein, the novel V-doped Ni3S2/NiS heterostructure nanorod arrays grown on nickel foam (VNS/NF-WM) are synthesized via a facile methanol-assisted hydrothermal method. We demonstrate that the morphology, phase composition, and crystallinity of VNS/NF are well modulated by tuning the ratios of water/methanol solvent. The optimized VNS/NF-WM-heterostructured nanorod (volume ratio of water/methanol is 3:1) exhibits superior HER electrocatalytic activity with low overpotentials of 85 and 218 mV to yield current density values of 10 and 100 mA cm-2, respectively, meanwhile sustaining an excellent stability with almost an unchanged current density of 10 mA cm-2 for 60 h. Our work offers fresh insights into the rational design of highly active and stable earth-abundant-heterostructured electrocatalysts for the hydrogen fuel production.

A mechanism for expansion of regulatory T-cell repertoire and its role in self-tolerance.

T-cell receptor (TCR) signalling has a key role in determining T-cell fate. Precursor cells expressing TCRs within a certain low-affinity range for complexes of self-peptide and major histocompatibility complex (MHC) undergo positive selection and differentiate into naive T cells expressing a highly diverse self-MHC-restricted TCR repertoire. In contrast, precursors displaying TCRs with a high affinity for 'self' are either eliminated through TCR-agonist-induced apoptosis (negative selection) or restrained by regulatory T (Treg) cells, whose differentiation and function are controlled by the X-chromosome-encoded transcription factor Foxp3 (reviewed in ref. 2). Foxp3 is expressed in a fraction of self-reactive T cells that escape negative selection in response to agonist-driven TCR signals combined with interleukin 2 (IL-2) receptor signalling. In addition to Treg cells, TCR-agonist-driven selection results in the generation of several other specialized T-cell lineages such as natural killer T cells and innate mucosal-associated invariant T cells. Although the latter exhibit a restricted TCR repertoire, Treg cells display a highly diverse collection of TCRs. Here we explore in mice whether a specialized mechanism enables agonist-driven selection of Treg cells with a diverse TCR repertoire, and the importance this holds for self-tolerance. We show that the intronic Foxp3 enhancer conserved noncoding sequence 3 (CNS3) acts as an epigenetic switch that confers a poised state to the Foxp3 promoter in precursor cells to make Treg cell lineage commitment responsive to a broad range of TCR stimuli, particularly to suboptimal ones. CNS3-dependent expansion of the TCR repertoire enables Treg cells to control self-reactive T cells effectively, especially when thymic negative selection is genetically impaired. Our findings highlight the complementary roles of these two main mechanisms of self-tolerance.

Cd271 mediates proliferation and differentiation of epidermal stem cells to support cutaneous burn wound healing.

Burn wounds can significantly reduce the quality of life of patients with respect to their physiology and psychology and can even threaten their lives. Many treatments have been proposed, including stem cell therapy but no effective method can as yet cure such damage. Our study highlights the role of Cd271 in epidermal stem cells (eSC) during the healing of burn wounds. The expression of Cd271 increases together with burn wound healing. Injection of Cd271-over-expressing eSC into wounds promotes the healing rate in a mouse burn model. Over-expression of Cd271 enhances the abilities of eSC with regard to their differentiation, proliferation and migration and even their resistance to apoptosis in vitro. These results are in accordance with a hypothesis suggesting that Cd271 promotes the healing of skin burn wounds by improving the potential of eSC for differentiation, proliferation and migration. Our findings shed light on the role of Cd271 in wound healing and may provide new therapeutic approaches for curing burn wounds of the skin.

Fullerenols Revisited: Highly Monodispersed Photoluminescent Nanomaterials as Ideal Building Blocks for Supramolecular Chemistry.

Fullerenols have been known for decades; however, the photoluminescent (PL) properties and applications in supramolecular chemistry of these molecules have not been well addressed. Herein, a strategy has been developed to purify the as-prepared fullerenols, and the photoluminescence and capabilities of these molecules to form supramolecular self-assemblies were systematically studied. It was found that fullerenols show wavelength-dependent emission with an absolute fluorescent quantum yield of approximately 3.5 %. The PL characteristics are reminiscent of carbon dots, especially those obtained by using the top-down method. The studied fullerenols can be used to detect Cu2+ ions with a limit of detection down to 3.50 µm. In addition, the amphiphilicity of the fullerenols can be readily tuned by ionic complexation with cationic surfactants, such as 1-tetradecyl-3-methylimidazoliumbromide (C14 mimB) and tetradecyltrimethylammonium bromide (TTAB). On increasing the concentration of the surfactant, the transition of aggregates was induced from highly ordered vesicles to honeycomb-structured crystals and finally to giant micelles. After the formation of supramolecular self-assemblies, enhancement of photoluminescence was observed, which can be ascribed to the suppression of intramolecular vibrations and motion combined with the loosely packed self-assembly array. This study provides a facile way to generate PL nanoarchitectures, which may find applications in fluorescent sensing, drug delivery, and optoelectronics.

A Novel Strategy for Improving the Aeromagnetic Compensation Performance of Helicopters.

An aeromagnetic survey is an important method in magnetic anomaly detection and geophysical prospecting. The magnetic field is typically measured by optically pumped magnetometers (OPM) installed on the aircraft. The measurement accuracy of the OPM is easily affected by the platform-generated magnetic fields. Therefore, aeromagnetic compensation is necessary. The traditional compensation model only considers the permanent, induced, and eddy current interference magnetic field of the aircraft platform. However, the interference field produced by the avionics system, and the relative motion between the aircraft and the magnetometer, are still not taken into account. To address this issue, we proposed a novel strategy to eliminate the additional interference of the platform with two OPMs. Among them, the OPM located farther away from the aircraft serves as a sensing magnetometer, whereas the near OPM serves as a reference magnetometer. The coherent noise suppression method is used to process the residual magnetic field interference after compensation. By establishing the interference magnetic transfer function between the two sensors, the interference field can be suppressed. The results of the experiments demonstrate the effectiveness of the novel strategy, and the standard deviation of residual interference drops from 0.065 nT to 0.045 nT.

Spectral Coefficient Analyses of Word-Initial Stop Consonant Productions Suggest Similar Anticipatory Coarticulation for Stuttering and Nonstuttering Adults.

A longstanding hypothesis about the sensorimotor mechanisms underlying stuttering suggests that stuttered speech dysfluencies result from a lack of coarticulation. Formant-based measures of either the stuttered or fluent speech of children and adults who stutter have generally failed to obtain compelling evidence in support of the hypothesis that these individuals differ in the timing or degree of coarticulation. Here, we used a sensitive acoustic technique-spectral coefficient analyses-that allowed us to compare stuttering and nonstuttering speakers with regard to vowel-dependent anticipatory influences as early as the onset burst of a preceding voiceless stop consonant. Eight adults who stutter and eight matched adults who do not stutter produced C1VC2 words, and the first four spectral coefficients were calculated for one analysis window centered on the burst of C1 and two subsequent windows covering the beginning of the aspiration phase. Findings confirmed that the combined use of four spectral coefficients is an effective method for detecting the anticipatory influence of a vowel on the initial burst of a preceding voiceless stop consonant. However, the observed patterns of anticipatory coarticulation showed no statistically significant differences, or trends toward such differences, between the stuttering and nonstuttering groups. Combining the present results for fluent speech in one given phonetic context with prior findings from both stuttered and fluent speech in a variety of other contexts, we conclude that there is currently no support for the hypothesis that the fluent speech of individuals who stutter is characterized by limited coarticulation.

Entropy and Entransy Dissipation Analysis of a Basic Organic Rankine Cycles (ORCs) to Recover Low-Grade Waste Heat Using Mixture Working Fluids.

Mixture working fluids can reduce effectively energy loss at heat sources and heat sinks, and therefore enhance the organic Rankine cycle (ORC) performance. The entropy and entransy dissipation analyses of a basic ORC system to recover low-grade waste heat using three mixture working fluids (R245fa/R227ea, R245fa/R152a and R245fa/pentane) have been investigated in this study. The basic ORC includes four components: an expander, a condenser, a pump and an evaporator. The heat source temperature is 120 °C while the condenser temperature is 20 °C. The effects of four operating parameters (evaporator outlet temperature, condenser temperature, pinch point temperature difference, degree of superheat), as well as the mass fraction, on entransy dissipation and entropy generation were examined. Results demonstrated that the entransy dissipation is insensitive to the mass fraction of R245fa. The entropy generation distributions at the evaporator for R245/pentane, R245fa/R152a and R245fa/R227ea are in ranges of 66-74%, 68-80% and 66-75%, respectively, with the corresponding entropy generation at the condenser ranges of 13-21%, 4-17% and 11-21%, respectively, while those at the expander for R245/pentane, R245fa/R152a and R245fa/R227ea are approaching 13%, 15% and 14%, respectively. The optimal mass fraction of R245fa for the minimum entropy generation is 0.6 using R245fa/R152a.

Whole-genome sequencing of cultivated and wild peppers provides insights into Capsicum domestication and specialization.

As an economic crop, pepper satisfies people's spicy taste and has medicinal uses worldwide. To gain a better understanding of Capsicum evolution, domestication, and specialization, we present here the genome sequence of the cultivated pepper Zunla-1 (C. annuum L.) and its wild progenitor Chiltepin (C. annuum var. glabriusculum). We estimate that the pepper genome expanded ∼0.3 Mya (with respect to the genome of other Solanaceae) by a rapid amplification of retrotransposons elements, resulting in a genome comprised of ∼81% repetitive sequences. Approximately 79% of 3.48-Gb scaffolds containing 34,476 protein-coding genes were anchored to chromosomes by a high-density genetic map. Comparison of cultivated and wild pepper genomes with 20 resequencing accessions revealed molecular footprints of artificial selection, providing us with a list of candidate domestication genes. We also found that dosage compensation effect of tandem duplication genes probably contributed to the pungent diversification in pepper. The Capsicum reference genome provides crucial information for the study of not only the evolution of the pepper genome but also, the Solanaceae family, and it will facilitate the establishment of more effective pepper breeding programs.

Signal transduction by the Fat cytoplasmic domain.

The large atypical cadherin Fat is a receptor for both Hippo and planar cell polarity (PCP) pathways. Here we investigate the molecular basis for signal transduction downstream of Fat by creating targeted alterations within a genomic construct that contains the entire fat locus, and by monitoring and manipulating the membrane localization of the Fat pathway component Dachs. We establish that the human Fat homolog FAT4 lacks the ability to transduce Hippo signaling in Drosophila, but can transduce Drosophila PCP signaling. Targeted deletion of conserved motifs identifies a four amino acid C-terminal motif that is essential for aspects of Fat-mediated PCP, and other internal motifs that contribute to Fat-Hippo signaling. Fat-Hippo signaling requires the Drosophila Casein kinase 1ε encoded by discs overgrown (Dco), and we characterize candidate Dco phosphorylation sites in the Fat intracellular domain (ICD), the mutation of which impairs Fat-Hippo signaling. Through characterization of Dachs localization and directed membrane targeting of Dachs, we show that localization of Dachs influences both the Hippo and PCP pathways. Our results identify a conservation of Fat-PCP signaling mechanisms, establish distinct functions for different regions of the Fat ICD, support the correlation of Fat ICD phosphorylation with Fat-Hippo signaling, and confirm the importance of Dachs membrane localization to downstream signaling pathways.

Apoptosis and death receptor signaling in diaphragm of burnt rats.

BACKGROUND: Respiratory dysfunction is a frequent complication after severe burn injury. Respiratory muscle atrophy may induce respiratory dysfunction due to insufficient inspiratory motive power. Accumulated evidence suggests that apoptosis is very important in skeletal muscle atrophy in multiple pathologic conditions. Therefore, we hypothesize that myonuclear apoptosis contributes to diaphragm atrophy induced by burn injury, and death receptor signaling activation plays a role in this process. METHODS: Wistar rats in the burn-injured group were subjected to a full-thickness scald injury around 40% of total body surface area. Diaphragm samples were examined for myonuclear apoptosis by transmission electron microscope, terminal deoxynucleotidyl transferase-mediated nick end labeling assay, and immunohistochemistry for caspase-3. Serum level of apoptotic ligands were assessed by ELISA. Activation of death receptor signaling was examined by Western blotting. RESULTS: Burn injury resulted in significant reductions of diaphragm muscle mass and myofiber cross-section area. Apoptosis in diaphragm appeared from day 1 and peaked on day 4 after injury. The level of soluble TNF-related apoptosis-inducing ligand and the ratio of Fas ligand to soluble Fas in serum significantly increased after burn injury. In diaphragm of burnt animals, the expressions of proapoptotic proteins, such as cleaved caspase-8, cleaved caspase-3, and Bax-to-Bcl-2 ratio were upregulated, whereas expression of pAkt, an antiapoptotic protein, was downregulated. Immunohistochemistry revealed that the most of the caspase-3 was expressed in myofiber nuclei and their surrounding cytoplasm area in tissue sections. CONCLUSIONS: Severe burn injury induces myonuclear apoptosis in diaphragm, which could be a contributor to diaphragm muscle atrophy. Activation of death receptor signaling may be a mechanism of apoptosis in diaphragm.