Spectratyping analysis of the islet-reactive T cell repertoire in diabetic NOD Igμnull mice after polyclonal B cell reconstitution

<p>Abstract</p> <p>Background</p> <p>Non Obese Diabetic mice lacking B cells (NOD.Igμ^nullmice) do not develop diabetes despite their susceptible background. Upon reconstitution of B cells using a chimera approach, animals start developing diabetes at 20 weeks of age.</p> <p>Methods</p> <p>We have used the spectratyping technique to follow the T cell receptor (TCR) V beta repertoire of NOD.Igμ^nullmice following B cell reconstitution. This technique provides an unbiased approach to understand the kinetics of TCR expansion. We have also analyzed the TCR repertoire of reconstituted animals receiving cyclophosphamide treatment and following tissue transplants to identify common aggressive clonotypes.</p> <p>Results</p> <p>We found that B cell reconstitution of NOD.Igμ^nullmice induces a polyclonal TCR repertoire in the pancreas 10 weeks later, gradually diversifying to encompass most BV families. Interestingly, these clonotypic BV expansions are mainly confined to the pancreas and are absent from pancreatic lymph nodes or spleens. Cyclophosphamide-induced diabetes at 10 weeks post-B cell reconstitution reorganized the predominant TCR repertoires by removing potential regulatory clonotypes (BV1, BV8 and BV11) and increasing the frequency of others (BV4, BV5S2, BV9, BV16-20). These same clonotypes are more frequently present in neonatal pancreatic transplants under the kidney capsule of B-cell reconstituted diabetic NOD.Igμ^nullmice, suggesting their higher invasiveness. Phenotypic analysis of the pancreas-infiltrating lymphocytes during diabetes onset in B cell reconstituted animals show a predominance of CD19⁺B cells with a B:T lymphocyte ratio of 4:1. In contrast, in other lymphoid organs (pancreatic lymph nodes and spleens) analyzed by FACS, the B:T ratio was 1:1. Lymphocytes infiltrating the pancreas secrete large amounts of IL-6 and are of Th1 phenotype after CD3-CD28 stimulation <it>in vitro</it>.</p> <p>Conclusions</p> <p>Diabetes in NOD.Igμ^nullmice appears to be caused by a polyclonal repertoire of T cell accumulation in pancreas without much lymphoid organ involvement and is dependent on the help by B cells.</p

Identification and Development of Synovial B-Cell-Related Genes Diagnostic Signature for Rheumatoid Arthritis

Background. The aim of the study was to investigate the landscape of B-cell-related gene expression profiling in rheumatoid arthritis (RA) synovium and explore the biological and clinical significance of these genes in RA. Methods. Expression profiling of synovial biopsies from subjects with 152 RA patients, 22 osteoarthritis (OA) patients, and 28 healthy controls was downloaded from the Gene Expression Omnibus database. Single-sample gene set enrichment analysis (ssGSEA) was performed to evaluate the abundance of infiltrated immune cells, and the results were validated using immunohistochemical staining. GSEA was employed to decipher differences in B-cell-related biological pathways. B-cell-related differential expression genes (BRDEGs) were screened, and BRDEGs-based model was developed by machine learning algorithms and evaluated by an external validation set and clinical RA cohort, then biological functions were further analyzed. Results. High levels of immune cell infiltration and B-cell-related pathway activation were revealed in RA synovium. BRDEGs were screened, and three key molecular markers consisting of FAS, GPR183, and TFRC were identified. The diagnosis model was established, and these gene markers have good discriminative ability for RA. Molecular pathological evaluation confirmed RA patients with high-risk scores presented higher levels of B-cell activation and RA characteristics. In addition, a competitive endogenous RNA network was established to elucidate the molecular mechanisms of the posttranscriptional network. Conclusions. We described the B-cell-related molecular landscape of RA synovium and constructed a molecular diagnostic model in RA. The three genes FAS, GPR183, and TFRC may be potential targets for clinical diagnosis and immunoregulatory therapy of RA

MicroRNA miR-509-3p inhibit metastasis and epithelial-mesenchymal transition in hepatocellular carcinoma

Our study seeks to obtain data which help to assess the impacts and related mechanisms of microRNA miR-509-3p in hepatocellular carcinoma (HCC). We found that the expression of miR-509-3p was down-regulated and Twist was up-regulated in HCC tissues and cell lines (HepG2, HCCLM3, Bel7402, and SMMC7721) compared with the adjacent normal tissues and normal human hepatocyte (L02). Moreover, cell proliferation, invasion, migration and epithelial–mesenchymal transition (EMT) in HepG2 and HCCLM3 cells were appeared to be markedly suppressed by overexpressed miR-509-3p. Overexpression of miR-509-3p also performed inhibition of the growth and metastasis in vivo. In addition, miR-509-3p could target and inhibit Twist expression, and it could further reverse the tumor promotion by Twist in HCC. All in all, miR-509-3p overexpression causes inhibition of the proliferation, migration, invasion and EMT of HCC cells by negatively regulating Twist, thereby suppressing HCC development and metastasis

Metagenomic next-generation sequencing reveals co-infection with Legionella pneumophila and Fusobacterium necrophorum in a patient with severe pneumonia: a case report

Abstract Background Legionella pneumonia is one of the most severe types of atypical pneumonia, impairing multiple organ systems, posing a threat to life. Diagnosing Legionella pneumonia is challenging due to difficulties in culturing the bacteria and limitations in immunoassay sensitivity and specificity. Case presentation This paper reports a rare case of sepsis caused by combined infection with Legionella pneumophila and Fusobacterium necrophorum, leading to respiratory failure, acute kidney injury, acute liver injury, myocardial damage, and electrolyte disorders. In addition, we systematically reviewed literature on patients with combined Legionella infections, analyzing their clinical features, laboratory results and diagnosis. Conclusions For pathogens that require prolonged incubation periods and are less sensitive to conventional culturing methods, metagenomic next-generation sequencing (mNGS) can be a powerful supplement to pathogen screening and plays a significant role in the auxiliary diagnosis of complex infectious diseases

A New Hybrid Decision-Making Model for Promoting Sustainable Social Rental Housing

The realization of sustainable social rental housing is regarded as an important policy to solve the housing burden, but social rental housing is often unsustainable. This study assesses the sustainability of social rental housing. However, the decision-making models, such as the classical decision-making hierarchy (AHP) used in the current study, conflict with the resolution of the complex influence of the relationship between evaluation dimensions/criteria. Based on this, we construct a new hybrid multi-attribute decision-making model (MADM) combining the Fuzzy Decision-Making Trial and Evaluation Laboratory (DEMATEL), Fuzzy DEMATEL-based analytic network process (DANP) and the modified VIseKriterijumska Optimizacija I Kompromisno Resenje (VIKOR) method for the sustainability assessment of government-led social rental housing (SRH) using Taiwanese experts as the subject of the investigation. The Fuzzy DEMATEL found that economic sustainability (D1), as the core dimension, affects environmental (D3) and social sustainability (D2). Furthermore, criteria with the “cause” position, including the reasonable development model (C2), the provision of living facilities and services needed (C5), the adoption of environmentally friendly technology and equipment (C10), guaranteed rent and lease (C1), the acquisition of working skill and employment opportunities (C6) and space planning and design (C3), should receive more attention from decision makers because improving these can further enhance the sustainability of the overall scheme. Combined with the DANP weight and modified VIKOR, we also indicate that these criteria should be optimized to make the evaluation more sustainable for a selected social housing case in Taiwan. The research also further proposes management strategy implications

Structure and features of the complete chloroplast genome of Salix triandroides (Salicaceae)

AbstractSalix triandroides W. P. Fang, an indigenous small willow with high potential for heavy-metal soil remediation and levee protection, has been used for wetland ecological restoration for decades. However, studies on the evolution and genetic breeding of S. triandoides have been hindered by the lack of its genetic information. Here, we present the complete chloroplast genome of S. triandroides, which was assembled from Illumina sequencing data. The chloroplast genome of this species is circular, 155,683 bp in length and includes two inverted repeat regions (IRs) (27,490 bp) separated by a large single-copy (LSC) region (84,463 bp) and a small single-copy (SSC) region (16,240 bp). A total of 126 unique genes were identified, including 81 protein-coding genes, 37 tRNA genes and eight rRNA genes. Comparative analysis identified some hypervariable regions, with potential to be used as specific DNA barcodes or candidate markers for phylogenetic studies. Based on the sequences of the protein-coding genes, the phylogenetic analysis assigned 32 Salix species into two major clades and revealed that S. triandoides was a sister taxon to S. triandra. Our results provide a foundation for further molecular breeding of S. triandroides and insights into the evolution of Salix.Supplemental data for this article is available online at https://doi.org/10.1080/13102818.2021.2023326

Insulinoma-released exosomes or microparticles are immunostimulatory and can activate autoreactive T cells spontaneously developed in nonobese diabetic mice

Exosomes (EXO) are secreted intracellular microparticles that can trigger inflammation and induce Ag-specific immune responses. To test possible roles of EXO in autoimmunity, we isolated small microparticles, mainly EXO, from mouse insulinoma and examined their activities to stimulate the autoimmune responses in NOD mice, a model for human type 1 diabetes. We demonstrate that the EXO contains strong innate stimuli and expresses candidate diabetes autoantigens. They can induce secretion of inflammatory cytokines through a MyD88-dependent pathway, and activate purified APC and result in T cell proliferation. To address whether EXO or the secreted microparticles are possible autoimmune targets causing islet-specific inflammation, we monitored the T cell responses spontaneously developed in prediabetic NOD mice for their reactivity to the EXO, and compared this reactivity between diabetes-susceptible and -resistant congenic mouse strains. We found that older NOD females, which have advanced islet destruction, accumulated more EXO-reactive, IFN-γ-producing lymphocytes than younger females or age-matched males, and that pancreatic lymph nodes from the prediabetic NOD, but not from the resistant mice, were also enriched with EXO-reactive Th1 cells. In vivo, immunization with the EXO accelerates insulitis development in nonobese diabetes-resistant mice. Thus, EXO or small microparticles can be recognized by the diabetes-associated autoreactive T cells, supporting that EXO might be a possible autoimmune target and/or insulitis trigger in NOD or congenic mouse strains

Two Domains of Vimentin Are Expressed on the Surface of Lymph Node, Bone and Brain Metastatic Prostate Cancer Lines along with the Putative Stem Cell Marker Proteins CD44 and CD133

Vimentin was originally identified as an intermediate filament protein present only as an intracellular component in many cell types. However, this protein has now been detected on the surface of a number of different cancer cell types in a punctate distribution pattern. Increased vimentin expression has been indicated as an important step in epithelial-mesenchymal transition (EMT) required for the metastasis of prostate cancer. Here, using two vimentin-specific monoclonal antibodies (SC5 and V9 directed against the coil one rod domain and the C-terminus of the vimentin protein, respectively), we examined whether either of these domains would be displayed on the surface of three commonly studied prostate cancer cell lines isolated from different sites of metastases. Confocal analysis of LNCaP, PC3 and DU145 prostate cancer cell lines (derived from lymph node, bone or brain prostate metastases, respectively) demonstrated that both domains of vimentin are present on the surface of these metastatic cancer cell types. In addition, flow cytometric analysis revealed that vimentin expression was readily detected along with CD44 expression but only a small subpopulation of prostate cancer cells expressed vimentin and the putative stem cell marker CD133 along with CD44. Finally, Cowpea mosaic virus (CPMV) nanoparticles that target vimentin could bind and internalize into tested prostate cancer cell lines. These results demonstrate that at least two domains of vimentin are present on the surface of metastatic prostate cancer cells and suggest that vimentin could provide a useful target for nanoparticle- or antibody- cancer therapeutic agents directed against highly invasive cancer and/or stem cells

Vitamin D status among infants and children in Shanghai, China: A hospital‐based study

Abstract The variation in vitamin D status is still unclear. We aim to describe the vitamin D status among healthy infants and children in Shanghai (31° N latitude), one of the largest cities in China. We conducted a hospital‐based, 2‐year retrospective observational study and recruited children for health examination at the Tongren Hospital affiliated with Shanghai Jiao Tong University School of Medicine from January 2019 to December 2020. Serum 25‐hydroxyvitamin D (25(OH)D) levels were measured using an enzyme‐linked immunosorbent assay. A total of 6164 children aged 0–11 years were included. Of these, 94.4% of the serum 25(OH)D measurements at first assessment were within the range of 12–50 ng/mL. The median 25(OH)D level was 31.3 (IQR 25.6, 38.1) ng/mL, the percentages of 25(OH)D < 20 ng/mL and 25(OH)D < 30 ng/mL were 10.0% and 43.8%, respectively. Low vitamin D status (deficiency and insufficiency) differed significantly by age group (infants, toddlers, preschoolers, and schoolers) and seasonality (all p < .001), but not by gender. For the sub‐group (n = 855) of children with repeated assessments, their low 25(OH)D levels increased significantly whether after about a 7‐month (n = 351) or 12‐month (n = 504) interval, and the increments of median 25(OH)D levels were 8.1 ng/mL and 2.1 ng/mL respectively (p < .001). This study documents the vitamin D status in Shanghai, showing that low vitamin D status is common in infants and children and suggesting that the assessment of 25(OH)D level is necessary for individuals who are at risk for deficiency or excess