RESUMEN
B cells can present antigens to CD4+ T cells, but it is thought that dendritic cells (DCs) are the primary initiators of naive CD4+ T cell responses. Nanoparticles, including virus-like particles (VLPs), are attractive candidates as carriers for vaccines and drug delivery. Using RNA phage Qß-derived VLP (Qß-VLP) as a model antigen, we found that antigen-specific B cells were the dominant antigen-presenting cells that initiated naive CD4+ T cell activation. B cells were sufficient to induce T follicular helper cell development in the absence of DCs. Qß-specific B cells promoted CD4+ T cell proliferation and differentiation via cognate interactions and through Toll-like receptor signaling-mediated cytokine production. Antigen-specific B cells were also involved in initiating CD4+ T cell responses during immunization with inactivated influenza virus. These findings have implications for the rational design of nanoparticles as vaccine candidates, particularly for therapeutic vaccines that aim to break immune tolerance.
Asunto(s)
Células Presentadoras de Antígenos/inmunología , Linfocitos B/inmunología , Linfocitos T CD4-Positivos/inmunología , Inmunización/métodos , Vacunas contra la Influenza/inmunología , Animales , Presentación de Antígeno/inmunología , Antígenos Virales/química , Antígenos Virales/inmunología , Diferenciación Celular/inmunología , Citocinas/inmunología , Citocinas/metabolismo , Subtipo H1N1 del Virus de la Influenza A/inmunología , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Nanopartículas/química , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/metabolismo , Receptores Toll-Like/inmunología , Vacunas de Productos Inactivados/inmunologíaRESUMEN
Zika virus (ZIKV) infection caused neurological complications and male infertility, leading to the accumulation of antigen-specific immune cells in immune-privileged organs (IPOs). Thus, it is important to understand the immunological responses to ZIKV in IPOs. We extensively investigated the ZIKV-specific T cell immunity in IPOs in Ifnar1-/- mice, based on an immunodominant epitope E294-302 tetramer. The distinct kinetics and functions of virus-specific CD8+ T cells infiltrated into different IPOs were characterized, with late elevation in the brain and spinal cord. Single epitope E294-302-specific T cells can account for 20-60% of the total CD8+ T cells in the brain, spinal cord, and testicle and persist for at least 90 days in the brain and spinal cord. The E294-302-specific TCRαßs within the IPOs are featured with the majority of clonotypes utilizing TRAV9N-3 paired with diverse TRBV chains, but with distinct αß paired clonotypes in 7 and 30 days post-infection. Specific chemokine receptors, Ccr2 and Ccr5, were selectively expressed in the E294-302-specific CD8+ T cells within the brain and testicle, indicating an IPO-oriented migration of virus-specific CD8+ T cells after infection. Overall, this study adds to the understanding of virus-specific CD8+ T cell responses for controlling and clearing ZIKV infection in IPOs.IMPORTANCEThe immune-privileged organs (IPOs), such as the central nervous system and testicles, presented pathogenicity and inflammation after Zika virus (ZIKV) infection with infiltrated CD8+ T cells. Our data show that CD8+ T cells keep up with virus increases and decreases in immune-privileged organs. Furthermore, our study provides the first ex vivo comparative analyses of the composition and diversity related to TCRα/ß clonotypes across anatomical sites and ZIKV infection phases. We show that the vast majority of TCRα/ß clonotypes in tissues utilize TRAV9N-3 with conservation. Specific chemokine expression, including Ccr2 and Ccr5, was found to be selectively expressed in the E294-302-specific CD8+ T cells within the brain and testicle, indicating an IPO-oriented migration of the virus-specific CD8+ T cells after the infection. Our study adds insights into the anti-viral immunological characterization and chemotaxis mechanism of virus-specific CD8+ T cells after ZIKV infection in different IPOs.
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Linfocitos T CD8-positivos , Privilegio Inmunológico , Infección por el Virus Zika , Animales , Masculino , Ratones , Encéfalo/inmunología , Encéfalo/virología , Linfocitos T CD8-positivos/inmunología , Receptor de Interferón alfa y beta/genética , Virus Zika , Infección por el Virus Zika/inmunología , Ratones Noqueados , Testículo/inmunología , Testículo/virologíaRESUMEN
Nonsense-mediated RNA decay (NMD) is a highly conserved and selective RNA turnover pathway that depends on the endonuclease SMG6. Here, we show that SMG6 is essential for male germ cell differentiation in mice. Germ-cell conditional knockout (cKO) of Smg6 induces extensive transcriptome misregulation, including a failure to eliminate meiotically expressed transcripts in early haploid cells, and accumulation of NMD target mRNAs with long 3' untranslated regions (UTRs). Loss of SMG6 in the male germline results in complete arrest of spermatogenesis at the early haploid cell stage. We find that SMG6 is strikingly enriched in the chromatoid body (CB), a specialized cytoplasmic granule in male germ cells also harboring PIWI-interacting RNAs (piRNAs) and the piRNA-binding protein PIWIL1. This raises the possibility that SMG6 and the piRNA pathway function together, which is supported by several findings, including that Piwil1-KO mice phenocopy Smg6-cKO mice and that SMG6 and PIWIL1 co-regulate many genes in round spermatids. Together, our results demonstrate that SMG6 is an essential regulator of the male germline transcriptome, and highlight the CB as a molecular platform coordinating RNA regulatory pathways to control sperm production and fertility.
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Endorribonucleasas , Gránulos de Ribonucleoproteína de Células Germinales , Espermatogénesis , Transcriptoma , Animales , Masculino , Ratones , Células Germinativas/metabolismo , ARN Interferente Pequeño/genética , Espermátides/metabolismo , Espermatogénesis/genética , Endorribonucleasas/metabolismoRESUMEN
Odontoblastic differentiation of human stem cells from the apical papilla (hSCAPs) is crucial for continued root development and dentin formation in immature teeth with apical periodontitis (AP). Fat mass and obesity-associated protein (FTO) has been reported to regulate bone regeneration and osteogenic differentiation profoundly. However, the effect of FTO on hSCAPs remains unknown. This study aimed to identify the potential function of FTO in hSCAPs' odontoblastic differentiation under normal and inflammatory conditions and to investigate its underlying mechanism preliminarily. Histological staining and micro-computed tomography were used to evaluate root development and FTO expression in SD rats with induced AP. The odontoblastic differentiation ability of hSCAPs was assessed via alkaline phosphatase and alizarin red S staining, qRT-PCR, and Western blotting. Gain- and loss-of-function assays and online bioinformatics tools were conducted to explore the function of FTO and its potential mechanism in modulating hSCAPs differentiation. Significantly downregulated FTO expression and root developmental defects were observed in rats with AP. FTO expression notably increased during in vitro odontoblastic differentiation of hSCAPs, while lipopolysaccharide (LPS) inhibited FTO expression and odontoblastic differentiation. Knockdown of FTO impaired odontoblastic differentiation, whereas FTO overexpression alleviated the inhibitory effects of LPS on differentiation. Furthermore, FTO promoted the expression of secreted modular calcium-binding protein 2 (SMOC2), and the knockdown of SMOC2 in hSCAPs partially attenuated the promotion of odontoblastic differentiation mediated by FTO overexpression under LPS-induced inflammation. This study revealed that FTO positively regulates the odontoblastic differentiation ability of hSCAPs by promoting SMOC2 expression. Furthermore, LPS-induced inflammation compromises the odontoblastic differentiation of hSCAPs by downregulating FTO, highlighting the promising role of FTO in regulating hSCAPs differentiation under the inflammatory microenvironment.
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Lipopolisacáridos , Osteogénesis , Humanos , Animales , Ratas , Ratas Sprague-Dawley , Microtomografía por Rayos X , Inflamación/genética , Proteínas de Unión al Calcio , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato/genéticaRESUMEN
Presynaptic increase in striatal dopamine is the primary dopaminergic abnormality in schizophrenia, but the underlying mechanisms are not understood. Here, we hypothesized that increased expression of endogenous GDNF could induce dopaminergic abnormalities that resemble those seen in schizophrenia. To test the impact of GDNF elevation, without inducing adverse effects caused by ectopic overexpression, we developed a novel in vivo approach to conditionally increase endogenous GDNF expression. We found that a 2-3-fold increase in endogenous GDNF in the brain was sufficient to induce molecular, cellular, and functional changes in dopamine signalling in the striatum and prefrontal cortex, including increased striatal presynaptic dopamine levels and reduction of dopamine in prefrontal cortex. Mechanistically, we identified adenosine A2a receptor (A2AR), a G-protein coupled receptor that modulates dopaminergic signalling, as a possible mediator of GDNF-driven dopaminergic abnormalities. We further showed that pharmacological inhibition of A2AR with istradefylline partially normalised striatal GDNF and striatal and cortical dopamine levels in mice. Lastly, we found that GDNF levels are increased in the cerebrospinal fluid of first episode psychosis patients, and in post-mortem striatum of schizophrenia patients. Our results reveal a possible contributor for increased striatal dopamine signalling in a subgroup of schizophrenia patients and suggest that GDNF-A2AR crosstalk may regulate dopamine function in a therapeutically targetable manner.
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Dopamina , Esquizofrenia , Animales , Ratones , Dopamina/metabolismo , Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Factor Neurotrófico Derivado de la Línea Celular Glial/farmacología , Esquizofrenia/metabolismo , Cuerpo Estriado/metabolismo , Transducción de SeñalRESUMEN
Tooth development is a complex process that is tightly controlled by circadian rhythm. Melatonin (MT) is a major hormonal regulator of the circadian rhythm, and influences dentin formation and odontoblastic differentiation during tooth development; however, the underlying mechanism remains elusive. This study investigated how MT regulates odontoblastic differentiation, with a special focus on its regulation of mitochondrial dynamics. In rat dental papilla cells (DPCs), we found that MT promotes odontoblastic differentiation concurrently with enhanced mitochondrial fusion, while disruption of mitochondrial fusion by depleting optic atrophy 1 (OPA1) impairs MT-mediated differentiation and mitochondrial respiratory functions. Through RNA sequencing, we discovered that MT significantly upregulated malic enzyme 2 (ME2), a mitochondrial NAD(P)+ -dependent enzyme, and identified ME2 as a critical MT downstream effector that orchestrates odontoblastic differentiation, mitochondrial fusion, and respiration functions. By detecting the spatiotemporal expression of ME2 in developing tooth germs, and using tooth germ reconstituted organoids, we also provided in vivo and ex vivo evidence that ME2 promotes dentin formation, indicating a possible involvement of ME2 in MT-modulated tooth development. Collectively, our findings offer novel understandings regarding the molecular mechanism by which MT affects cell differentiation and organogenesis, meanwhile, the critical role of ME2 in MT-regulated mitochondrial functions is also highlighted.
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Melatonina , Animales , Ratas , Diferenciación Celular , Pulpa Dental , Melatonina/metabolismo , Dinámicas Mitocondriales , Odontoblastos/metabolismo , Respiración , Malato Deshidrogenasa/metabolismoRESUMEN
The emergence of Transformer has led to the rapid development of video understanding, but it also brings the problem of high computational complexity. Previously, there were methods to divide the feature maps into windows along the spatiotemporal dimensions and then calculate the attention. There are also methods to perform down-sampling during attention computation to reduce the spatiotemporal resolution of features. Although the complexity is effectively reduced, there is still room for further optimization. Thus, we present the Windows and Linear Transformer (WLiT) for efficient video action recognition, by combining Spatial-Windows attention with Linear attention. We first divide the feature maps into multiple windows along the spatial dimensions and calculate the attention separately inside the windows. Therefore, our model further reduces the computational complexity compared with previous methods. However, the perceptual field of Spatial-Windows attention is small, and global spatiotemporal information cannot be obtained. To address this problem, we then calculate Linear attention along the channel dimension so that the model can capture complete spatiotemporal information. Our method achieves better recognition accuracy with less computational complexity through this mechanism. We conduct extensive experiments on four public datasets, namely Something-Something V2 (SSV2), Kinetics400 (K400), UCF101, and HMDB51. On the SSV2 dataset, our method reduces the computational complexity by 28% and improves the recognition accuracy by 1.6% compared to the State-Of-The-Art (SOTA) method. On the K400 and two other datasets, our method achieves SOTA-level accuracy while reducing the complexity by about 49%.
RESUMEN
Few-shot object detection (FSOD) is proposed to solve the application problem of traditional detectors in scenarios lacking training samples. The meta-learning methods have attracted the researchers' attention for their excellent generalization performance. They usually select the same class of support features according to the query labels to weight the query features. However, the model cannot possess the ability of active identification only by using the same category support features, and feature selection causes difficulties in the testing process without labels. The single-scale feature of the model also leads to poor performance in small object detection. In addition, the hard samples in the support branch impact the backbone's representation of the support features, thus impacting the feature weighting process. To overcome these problems, we propose a multi-scale feature fusion and attentive learning (MSFFAL) framework for few-shot object detection. We first design the backbone with multi-scale feature fusion and channel attention mechanism to improve the model's detection accuracy on small objects and the representation of hard support samples. Based on this, we propose an attention loss to replace the feature weighting module. The loss allows the model to consistently represent the objects of the same category in the two branches and realizes the active recognition of the model. The model no longer depends on query labels to select features when testing, optimizing the model testing process. The experiments show that MSFFAL outperforms the state-of-the-art (SOTA) by 0.7-7.8% on the Pascal VOC and exhibits 1.61 times the result of the baseline model in MS COCO's small objects detection.
RESUMEN
KEY MESSAGE: A novel major QTL for FHB resistance was mapped to a 6.8 Mb region on chromosome 2D in a Chinese wheat cultivar Ji5265, and diagnostic KASP markers were developed for detecting it in a worldwide wheat collection. Fusarium head blight (FHB) is a serious disease in wheat (Triticum aestivum L.) and causes significant reductions in grain yield and quality worldwide. Breeding for FHB resistance is the most effective strategy to minimize the losses caused by FHB; therefore, identification of major quantitative trait loci (QTLs) conferring FHB resistance and development of diagnostic markers for the QTLs are prerequisites for marker-assisted selection (MAS). Ji5265 is a Chinese wheat cultivar resistant to FHB in multiple environments. An F6 population of 179 recombinant inbred lines (RILs) was developed from Ji5265 × Wheaton. The population was genotyped by genotyping-by-sequencing (GBS) and phenotyped for FHB Type II resistance in greenhouses. A major QTL, designated as QFhb-2DL, was mapped in a 6.8 Mb region between the markers GBS10238 and GBS12056 on the long arm of chromosome 2D in Ji5265 and explained ~ 30% of the phenotypic variation for FHB resistance. The effect of QFhb-2DL on FHB resistance was validated using near-isogenic lines (NILs) derived from residual heterozygotes from an F6 RIL of Ji5265 × Wheaton. The two flanking markers were converted into Kompetitive allele-specific PCR (KASP) markers (KASP10238 and KASP12056) and validated to be diagnostic in a collection of 2,065 wheat accessions. These results indicate that QFhb-2DL is a novel major QTL for resistance to FHB spread within a spike (Type II) and the two KASP markers can be used for MAS to improve wheat FHB resistance in wheat breeding programs.
Asunto(s)
Fusarium , China , Mapeo Cromosómico , Fitomejoramiento , Enfermedades de las Plantas/genética , Sitios de Carácter Cuantitativo , Triticum/genéticaRESUMEN
T cell antigen receptor (TCR) signaling is essential for the differentiation and maintenance of effector regulatory T (Treg) cells. However, the contribution of individual TCR-dependent genes in Treg cells to the maintenance of immunotolerance remains largely unknown. Here we demonstrate that Treg cells lacking E protein undergo further differentiation into effector cells that exhibit high expression of effector Treg signature genes, including IRF4, ICOS, CD103, KLRG-1, and RORγt. E protein-deficient Treg cells displayed increased stability and enhanced suppressive capacity. Transcriptome and ChIP-seq analyses revealed that E protein directly regulates a large proportion of the genes that are specific to effector Treg cell activation, and importantly, most of the up-regulated genes in E protein-deficient Treg cells are also TCR dependent; this indicates that E proteins comprise a critical gene regulatory network that links TCR signaling to the control of effector Treg cell differentiation and function.
Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Diferenciación Celular/inmunología , Receptores de Antígenos de Linfocitos T/inmunología , Transducción de Señal , Linfocitos T Reguladores/inmunología , Animales , Redes Reguladoras de Genes , Homeostasis , Ratones , Linfocitos T Reguladores/citologíaRESUMEN
Accurate and reliable stride length estimation modules play a significant role in Pedestrian Dead Reckoning (PDR) systems, but the accuracy of stride length calculation suffers from individual differences. This paper presents a stride length prediction strategy for PDR systems that can be adapted across individuals and broad walking velocity fields. It consists of a multi-gait division algorithm, which can divide a full stride into push-off, swing, heel-strike, and stance based on multi-axis IMU data. Additionally, based on the acquired gait phases, the correlation between multiple features of distinct gait phases and the stride length is analyzed, and multi regression models are merged to output the stride length value. In experimental tests, the gait segmentation algorithm provided gait phases division with the F-score of 0.811, 0.748, 0.805, and 0.819 for stance, push-off, swing, heel-strike, respectively, and IoU of 0.482, 0.69, 0.509 for push-off, swing, heel-strike, respectively. The root means square error (RMSE) of our proposed stride length estimation was 151.933, and the relative error for total distance in varying walking speed tests was less than 2%. The experimental results validated that our proposed gait phase segmentation algorithm can accurately recognize gait phases for individuals with wide walking speed ranges. With no need for parameter modification, the stride length method based on the fusion of multiple predictions from different gait phases can provide better accuracy than the estimations based on the full stride.
Asunto(s)
Marcha , Peatones , Talón , Humanos , Caminata , Velocidad al CaminarRESUMEN
Stride length estimation is one of the most crucial aspects of Pedestrian Dead Reckoning (PDR). Due to the measurement noise of inertial sensors, individual variances of pedestrians, and the uncertainty in pedestrians walking, there is a substantial error in the assessment of stride length, which causes the accumulated deviation of Pedestrian Dead Reckoning (PDR). With the help of multi-gait analysis, which decomposes strides in time and space with greater detail and accuracy, a novel and revolutionary stride estimating model or scheme could improve the performance of PDR on different users. This paper presents a diverse stride gait dataset by using inertial sensors that collect foot movement data from people of different genders, heights, and walking speeds. The dataset contains 4690 walking strides data and 19,083 gait labels. Based on the dataset, we propose a threshold-independent stride segmentation algorithm called SDATW and achieve an F-measure of 0.835. We also provide the detailed results of recognizing four gaits under different walking speeds, demonstrating the utility of our dataset for helping train stride segmentation algorithms and gait detection algorithms.
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Peatones , Algoritmos , Femenino , Marcha , Análisis de la Marcha , Humanos , Masculino , Caminata , Velocidad al CaminarRESUMEN
Long-chain noncoding RNA (lncRNA) is a new class of molecular regulators in heart development and disease. However, the role of specific lncRNA in cardiac fibrosis remains to be fully explored. This study aimed to investigate the role and potential mechanism of lncRNA MHRT in myocardial fibrosis after myocardial infarction (MI).Cardiac fibroblasts (CFs) were isolated from a mouse model of MI. The expression levels of MHRT and miR-3185 in the hearts of MI and CFs mice treated with transforming growth factor beta 1 (TGF-ß1) were analyzed by qRT-PCR. The collagen expression was assessed using qRT-PCR and Western blot. Cell proliferation was assessed by performing MTT and EdU assays. The direct interaction between lncRNA and miRNA was analyzed by luciferase assay, RNA-binding protein immunoprecipitation (RIP) assay, and RNA pull-down assay.The expression levels of MHRT were raised in MI and CFs mice treated with TGF-ß1. Overexpression of MHRT promoted collagen production and CF proliferation, while silencing of MHRT showed the opposite effect. MiR-3185 was a target gene of MHRT. In addition, overexpression of MHRT reduced the expression levels of miR-3185, and siMHRT reversed the inhibitory effect of TGF-ß1 on the expression of miR-3185. Overexpression of miR-3185 inhibited the upregulation of Col I and Col III induced by TGF-ß1.MHRT promoted cardiac fibrosis after MI through miR-3185 and increased myocardial collagen deposition and promoted myocardial fibrosis.
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Infarto del Miocardio/metabolismo , Miocardio/patología , ARN Largo no Codificante/metabolismo , Animales , Colágeno/metabolismo , Fibroblastos/metabolismo , Fibrosis , Masculino , Ratones Endogámicos C57BL , Infarto del Miocardio/patología , Ratas Sprague-DawleyRESUMEN
Dental papilla cells (DPCs), precursors of odontoblasts, are considered promising seed cells for tissue engineering. Emerging evidence suggests that melatonin promotes odontoblastic differentiation of DPCs and affects tooth development, although the precise mechanisms remain unknown. Retinoid acid receptor-related orphan receptor α (RORα) is a nuclear receptor for melatonin that plays a critical role in cell differentiation and embryonic development. This study aimed to explore the role of RORα in odontoblastic differentiation and determine whether melatonin exerts its pro-odontogenic effect via RORα. Herein, we observed that RORα was expressed in DPCs and was significantly increased during odontoblastic differentiation in vitro and in vivo. The overexpression of RORα upregulated the expression of odontogenic markers, alkaline phosphatase (ALP) activity and mineralized nodules formation (p < 0.05). In contrast, odontoblastic differentiation of DPCs was suppressed by RORα knockdown. Moreover, we found that melatonin elevated the expression of odontogenic markers, which was accompanied by the upregulation of RORα (p < 0.001). Utilising small interfering RNA, we further demonstrated that RORα inhibition attenuated melatonin-induced odontogenic gene expression, ALP activity and matrix mineralisation (p < 0.01). Collectively, these results provide the first evidence that RORα can promote odontoblastic differentiation of DPCs and mediate the pro-odontogenic effect of melatonin.
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Diferenciación Celular , Papila Dental/citología , Melatonina/farmacología , Miembro 1 del Grupo F de la Subfamilia 1 de Receptores Nucleares/metabolismo , Odontoblastos/citología , Odontoblastos/metabolismo , Odontogénesis , Animales , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Odontoblastos/efectos de los fármacos , Odontogénesis/efectos de los fármacos , Ratas Sprague-Dawley , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Acute kidney injury (AKI) is a clinical condition that is associated with high morbidity and mortality. Inflammation is reported to play a key role in AKI. Although the M2 macrophages exhibit antimicrobial and anti-inflammatory activities, their therapeutic potential has not been evaluated for AKI. This study aimed to investigate the protective effect of peritoneal M2 macrophage transplantation on AKI in mice. The macrophages were isolated from peritoneal dialysates of mice. The macrophages were induced to undergo M2 polarization using interleukin (IL)-4/IL-13. AKI was induced in mice by restoring the blood supply after bilateral renal artery occlusion for 30 minutes. The macrophages were injected into the renal cortex of mice. The changes in renal function, inflammation and tubular proliferation were measured. The M2 macrophages were co-cultured with the mouse primary proximal tubular epithelial cells (PTECs) under hypoxia/reoxygenation conditions in vitro. The PTEC apoptosis and proliferation were analysed. The peritoneal M2 macrophages effectively alleviated the renal injury and inflammatory response in mice with ischaemia-reperfusion injury (IRI) and promoted the PTEC proliferation in vivo and in vitro. These results indicated that the peritoneal M2 macrophages ameliorated AKI by decreasing inflammatory response and promoting PTEC proliferation. Hence, the peritoneal M2 macrophage transplantation can serve as a potential cell therapy for renal diseases.
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Lesión Renal Aguda/terapia , Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Macrófagos Peritoneales/trasplante , Daño por Reperfusión/terapia , Animales , Apoptosis/fisiología , Proliferación Celular/fisiología , Supervivencia Celular , Células Cultivadas , Técnicas de Cocultivo , Inflamación/patología , Riñón/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Obstrucción de la Arteria Renal , Cicatrización de Heridas/fisiologíaRESUMEN
Diabetic nephropathy (DN) as a kind of serious microvascular complication of Diabetes Mellitus (DM) usually causes the end-stage of renal disease (ESRD). Studies have demonstrated that CD103+ dendritic cells (DCs) exhibited a renal pathogenic effect in murine chronic kidney disease (CKD). Mesenchymal stem cells (MSCs) can alleviate DN and suppress the DCs maturation. To explore the role of CD103+ DCs and the potential mechanisms underlying MSCs-mediated protective effects in DN, we used bone marrow MSCs (BM-MSCs) to treat DN rats. MSCs transplantation considerably recovered kidney function and diminished renal injury, fibrosis and the population of renal CD103+ DCs in DN rat. The MSCs-treated DN rats had decreased mRNA expression levels of interleukin (IL)1ß, IL6, tumour necrosis factor alpha (TNF-α), monocyte chemotactic protein 1 (MCP-1) and reduced CD8 T cell infiltration in the kidney. MSCs significantly down-regulated the genes expression of transcription factors (Basic leucine zipper transcriptional factor ATF-like 3, Batf3 and DNA-binding protein inhibitor ID-2, Id2) and FMS-like tyrosine kinase-3 (Flt3) which are necessary for CD103+ DCs development. The protective effect of MSCs may be partly related to their immunosuppression of CD8+ T cell proliferation and activation mediated by CD103+ DCs in the kidney of DN rats.
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Antígenos CD/metabolismo , Linfocitos T CD8-positivos/inmunología , Células Dendríticas/metabolismo , Nefropatías Diabéticas/inmunología , Nefropatías Diabéticas/terapia , Cadenas alfa de Integrinas/metabolismo , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Animales , Antígenos de Diferenciación Mielomonocítica/metabolismo , Proliferación Celular , Citotoxicidad Inmunológica , Nefropatías Diabéticas/patología , Inflamación/patología , Riñón/lesiones , Riñón/patología , Activación de Linfocitos/inmunología , Macrófagos/metabolismo , Macrófagos/patología , Masculino , Modelos Biológicos , Ratas Sprague-DawleyRESUMEN
Hydroxysteroid 17ß dehydrogenase 12 (HSD17B12) is suggested to be involved in the elongation of very long chain fatty acids. Previously, we have shown a pivotal role for the enzyme during mouse development. In the present study we generated a conditional Hsd17b12 knockout (HSD17B12cKO) mouse model by breeding mice homozygous for a floxed Hsd17b12 allele with mice expressing the tamoxifen-inducible Cre recombinase at the ROSA26 locus. Gene inactivation was induced by administering tamoxifen to adult mice. The gene inactivation led to a 20% loss of body weight within 6 days, associated with drastic reduction in both white (83% males, 75% females) and brown (65% males, 60% females) fat, likely due to markedly reduced food and water intake. Furthermore, the knockout mice showed sickness behavior and signs of liver toxicity, specifically microvesicular hepatic steatosis and increased serum alanine aminotransferase (4.6-fold in males, 7.7-fold in females). The hepatic changes were more pronounced in females than males. Proinflammatory cytokines, such as interleukin-6 (IL-6), IL-17, and granulocyte colony-stimulating factor, were increased in the HSD17B12cKO mice indicating an inflammatory response. Serum lipidomics study showed an increase in the amount of dihydroceramides, despite the dramatic overall loss of lipids. In line with the proposed role for HSD17B12 in fatty acid elongation, we observed accumulation of ceramides, dihydroceramides, hexosylceramides, and lactosylceramides with shorter than 18-carbon fatty acid side chains in the serum. The results indicate that HSD17B12 is essential for proper lipid homeostasis and HSD17B12 deficiency rapidly results in fatal systemic inflammation and lipolysis in adult mice.
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17-Hidroxiesteroide Deshidrogenasas/fisiología , Homeostasis/fisiología , 17-Hidroxiesteroide Deshidrogenasas/genética , Tejido Adiposo Pardo/efectos de los fármacos , Tejido Adiposo Pardo/metabolismo , Tejido Adiposo Blanco/efectos de los fármacos , Tejido Adiposo Blanco/metabolismo , Animales , Conducta Animal , Peso Corporal/genética , Citocinas/metabolismo , Ácidos Grasos/metabolismo , Conducta Alimentaria , Femenino , Homeostasis/genética , Metabolismo de los Lípidos/genética , Metabolismo de los Lípidos/fisiología , Lipidómica , Hepatopatías/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Caracteres Sexuales , Tamoxifeno/farmacologíaRESUMEN
RNA editing is a post-transcriptional process of modifying genetic information on RNA molecules, which provides cells an additional level of gene expression regulation. Unlike mammals, in land plants, RNA editing converts C-to-U residues in organelles. However, its potential roles in response to different stressors (heat, salt, and so on) remains unclear. Grape is one of the most popular and economically important fruits in the world, and its production, like other crops, must deal with abiotic and biotic stresses, which cause reductions in yield and fruit quality. In our study, we tested the influence of the environmental factor temperature on RNA editing process in the whole mRNA from grape organelle. In total, we identified 122 and 627 RNA editing sites in chloroplast and mitochondria respectively with the average editing efficiency nearly ~ 60%. The analyses revealed that number of non-synonymous editing sites were higher than that of synonymous editing sites, and the amino acid substitution type tends to be hydrophobic. Additionally, the overall editing level decreased with the temperature rises, especially for several gene transcripts in chloroplast and mitochondria (matK, ndhB, etc.). We also found that the expression level of most PPR genes decreased with the temperature rises, which may contribute to the decline of RNA editing efficiency at high temperature. Our findings suggested that the RNA editing events were very sensitive to heat stress; the changes of amino acid in RNA editing genes may contribute to the stress adaption for grape.
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Edición de ARN , Termotolerancia , Vitis/genética , Cloroplastos/genética , Regulación de la Expresión Génica de las Plantas , Mitocondrias/genética , Vitis/fisiologíaRESUMEN
Antigen-specific Th1 cells could be a passage to the infection sites during infection to execute effector functions, such as help CD8+ T cells to localize in these sites by secretion of anti-viral cytokines-IFN-γ or direct cytotoxicity of antigen-bearing cells. However, the molecular components that modulate Th1 cell differentiation and function in response to viral infection remain incompletely understood. Here, we reported that both inhibitor of DNA binding 3(Id3) protein and inhibitor of DNA binding 2(Id2) protein promoted Th1 cell differentiation. Depletion of Id3 or Id2 led to severe defect of Th1 cell differentiation during influenza virus infection. Whereas depletion of both Id3 and Id2 in CD4+ T cells restrained Th1 cell differentiation to a greater extent, indicating that Id3 and Id2 nonredundantly regulate Th1 cell differentiation. Moreover, deletion of E-proteins, the antagonists of Id proteins, greatly enhanced Th1 cell differentiation. Mechanistic study indicated that E-proteins suppressed Th1 cell differentiation by directly binding to the regulatory elements of Th1 cell master regulator T-bet and regulate T-bet expression. Thus, our findings identified Id-protein's importance for Th1 cells and clarified the nonredundant role of Id3 and Id2 in regulating Th1 cell differentiation, providing novel insight that Id3-Id2-E protein axis are essential for Th1 cell polarization.
Asunto(s)
Diferenciación Celular/inmunología , Proteína 2 Inhibidora de la Diferenciación/inmunología , Proteínas Inhibidoras de la Diferenciación/inmunología , Infecciones por Orthomyxoviridae/inmunología , Orthomyxoviridae/inmunología , Células TH1/inmunología , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/inmunología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD4-Positivos/virología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/virología , Diferenciación Celular/genética , Regulación de la Expresión Génica/inmunología , Proteína 2 Inhibidora de la Diferenciación/genética , Proteína 2 Inhibidora de la Diferenciación/metabolismo , Proteínas Inhibidoras de la Diferenciación/genética , Proteínas Inhibidoras de la Diferenciación/metabolismo , Ratones Noqueados , Ratones Transgénicos , Orthomyxoviridae/fisiología , Infecciones por Orthomyxoviridae/genética , Infecciones por Orthomyxoviridae/virología , Elementos Reguladores de la Transcripción/genética , Proteínas de Dominio T Box/genética , Proteínas de Dominio T Box/inmunología , Proteínas de Dominio T Box/metabolismo , Células TH1/metabolismo , Células TH1/virologíaRESUMEN
Sperm differentiation requires specific protein transport for correct sperm tail formation and head shaping. A transient microtubular structure, the manchette, appears around the differentiating spermatid head and serves as a platform for protein transport to the growing tail. Sperm flagellar 2 (SPEF2) is known to be essential for sperm tail development. In this study we investigated the function of SPEF2 during spermatogenesis using a male germ cell-specific Spef2 knockout mouse model. In addition to defects in sperm tail development, we observed a duplication of the basal body and failure in manchette migration resulting in an abnormal head shape. We identified cytoplasmic dynein 1 and GOLGA3 as novel interaction partners for SPEF2. SPEF2 and dynein 1 colocalize in the manchette and the inhibition of dynein 1 disrupts the localization of SPEF2 to the manchette. Furthermore, the transport of a known SPEF2-binding protein, IFT20, from the Golgi complex to the manchette was delayed in the absence of SPEF2. These data indicate a possible novel role of SPEF2 as a linker protein for dynein 1-mediated cargo transport along microtubules.