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关键词： Photobiomodulation   Spinal cord injury   Autophagy   Inflammation   TLR2  

摘要： After spinal cord injury (SCI), peripherally derived macrophages infiltrated the injury area to exert inflammatory effects, causing barriers to the repair of spinal cord injury. Our previous study confirmed that photobiomodulation (PBM) could promote the motor function recovery and inhibit the secretion of inflammatory cytokines after SCI, moreover, PBM also has a role in promoting autophagy, but the mechanism is not clear. Therefore, we aimed to investigate whether PBM promotes autophagy by regulating the inflammatory response of macrophages, which in turn regulates functional repair after SCI. Male C57/BL6 mice were used to prepare a model of clamped spinal cord injury, and PBM irradiation was performed for 28 consecutive days, which showed that motor function of the mice was improved. We observed that autophagy proteins (LC3, Beclin-1 and P62) were inhibited and inflammasome-related proteins (NLRP3, Caspase-1 and IL-1 beta) expression was significantly enhanced in SCI mice. We analyzed the RNA sequencing (RNA-seq) of SCI, SCI+PBM treated mice in combination with autophagy database. The results showed 25 differentially expressed genes (DEGs). Protein-protein interaction (PPI) network and Hub gene analysis revealed TLR2 as a key molecule in the regulation of autophagy levels by PBM after SCI. We performed preliminary analysis in macrophages cultured in vitro and observed that PBM suppressed the expression of TLR2 and inflammasome-related proteins in M1-type macrophages and promoted the expression of autophagy proteins. Subsequently, we used an agonist of TLR2 (CU-T12-9) to up regulate TLR2 expression and observed that macrophage autophagy was inhibited and inflammatory response was enhanced. After PBM irradiation, the effect of CU-T12-9 was counteracted. Taken together, PBM promotes autophagy and attenuates the inflammatory response by regulating TLR2, a key molecule of autophagy in spinal cord injury.

关键词： TNF-alpha antagonist   Diabetic rat colon   Volume of colonic layers   Number of myenteric neurons   Death commitment points   Stereology   Physical disector  

摘要： Background: this study aimed to examine the impact of a TNF-alpha antagonist on markers of neuronal death, the total number of myenteric neurons, and the volume of each colonic layer in diabetic rats. Methods: thirty male rats were divided into normal control (NC), diabetic control (DC), and diabetic etanercept (DE) groups. Diabetes was induced using streptozotocin and nicotinamide. The DE group received twice-weekly injections of etanercept. Immunohistochemistry staining and western blotting detected activated caspase-3, Gasdermin D, and phosphorylated-MLKL. Stereological methods were performed in colon sections stained with Toluidine-blue to estimate the total number of neurons in the colonic myenteric plexus and the colonic layer volume. Results: immunohistochemistry showed etanercept reduced expression of apoptosis and pyroptosis markers, but not necroptosis, in the myenteric plexus of the colon in diabetic rats. Diabetic rats exhibited a lower total number of neurons in the colonic myenteric ganglions, and etanercept did not prevent this neuronal loss. Furthermore, colonic layer enlargement was observed in diabetic rats, with only the muscular layer hypertrophy being prevented by etanercept administration. Conclusions: the TNF-alpha antagonist did not prevent neuronal loss in the colonic myenteric plexus;it partially inhibited colonic muscular layer enlargement in diabetic rats.

关键词： CHO‐K1   IL‐6   IL‐6Rα   Tocilizumab   biosimilar   lentivirus   mAb  

摘要： Tocilizumab is a monoclonal antibody (mAb) that recognizes human Interleukin 6 receptor alpha (IL-6Rα) and antagonizes IL-6 signaling. It is therefore used in the treatment of moderate to severe rheumatoid arthritis in patients with inadequate response to antirheumatic drugs, and in the treatment of systemic juvenile idiopathic arthritis. During the course of the Coronavirus Disease 2019 (COVID-19) pandemic, the potential application of Tocilizumab in severe inflammatory diseases was demonstrated. In addition, the drug is attracting increasing attention for its potential application in other pathologies, particularly cancers where the central role of Interleukin 6 (IL-6) has been described, such as pancreatic ductal adenocarcinoma. The necessity for further research in this area is far too great, which is why the development of additional biosimilars is both necessary and welcome. In this work, we describe the generation and biological characterization of a biosimilar candidate of Tocilizumab. A stable producing CHO-K1 cell line was obtained by lentiviral transduction. We demonstrated that the antibody generated, named 41TB, has similar size heterogeneity and electrophoretic profile as Tocilizumab, and high levels of purity. Moreover, 41TB exhibits comparable binding capacity to IL-6Rα, as well as the ability to interrupt IL-6/IL-6Rα binding in ELISA. Additionally, both antibodies were likewise able to inhibit DS-1 cell proliferation, IL-6 induced STAT3 phosphorylation, and C-Reactive Protein production in vitro. Galactose supplementation of the culture medium appears to be necessary to generate a glycosylation profile similar to the original antibody.

关键词： hPSC   HLA   gene editing   immune checkpoint  

摘要： Human pluripotent stem cells (hPSCs) hold great promise in regenerative medicine. However, immune rejections remain one of the major obstacles to stem cell therapy. Though conventional immunosuppressants are available in clinics, the side effects prevent the wide application of hPSCs derivatives, compromising both survival rate and quality of life. In recent years, a myriad of strategies aimed at inducing immune tolerance specifically by engineering stem cells has been introduced to society. One strategy involves human leukocyte antigen (HLA) deletion through gene editing, affording allografts the capability to evade the host immune system. Another strategy involves immune cloak, which is the focus of this review, with emphasis on the overexpression of immune checkpoints and the blocking of immune cytotoxic pathways. Nevertheless, co-transplantation with mesenchymal stem cells (MSCs) and enhanced MSCs confers immune privilege to engraftments. This review summarizes recent studies on the intricacies of immune tolerance induction by engineering stem cells. In addition, we endeavor to deliberate upon the safety and limitations associated with this promising and potential therapeutic modality.

关键词： flavivirus   tick-borne encephalitis virus   NS1 protein   endothelial hyperpermeability   TNF-alpha signaling pathway   TEER  

摘要： Orthoflavivirus encephalitidis (tick-borne encephalitis virus, TBEV) is of high concern due to its ability to cause severe neurological manifestations. Despite the fact that the role of NS1 proteins from various mosquito-borne flaviviruses in pathogenesis and their ability to affect human endothelial permeability have been investigated, TBEV NS1 has thus far been insufficiently studied. In this study, human endothelial permeability was assessed using TEER and transwell permeability assays. Signaling pathways were determined by RNAseq. The ability of the NS1 protein of TBEV to affect human endothelial permeability was investigated for the first time. It was shown that recombinant TBEV NS1 produced in eucaryotic cells directly affected human lung microvascular endothelial cells (HLMVECs) in vitro but not human umbilical vein endothelial cells (HUVECs). It was indicated that TBEV NS1 induced endothelial hyperpermeability of HLMVECs through activating TNF-alpha and other inflammatory signaling pathways.

关键词： autoimmunity   anti-PD1-antibody   cutaneous side effects   skin   gene expression   bullous pemphigoid   immune-related bullous pemphigoid   immune-related adverse events   immune checkpoint inhibitor  

摘要： Background/Objectives: Cutaneous side effects are the most common immune-related adverse events (irAEs) caused by immune checkpoint inhibitors (ICIs) and affect 70-90% of patients. Besides diverse types of exanthema, rare skin toxicity includes bullous dermatoses in 0.3% of cases. Systemic steroids are the first-line treatment for immune-related bullous pemphigoid (irBP);however, some cases are corticosteroid-resistant. IrBP is one of the irAEs most frequently chronic and associated with long-term steroid use. However, steroids may interfere with tumor response. Therefore, alternative treatment strategies for irBP are desperately needed. Dupilumab, a monoclonal antibody blocking the receptor binding of interleukin-4 (IL-4) and interleukin-13 (IL-13), has been successfully used to treat spontaneous forms of bullous pemphigoid (BP). In this study, we analyzed the gene expression profiles of BP and irBP. Patients and Methods: A retrospective multicenter study evaluated the gene expression profiles of irBP and BP in comparison to healthy controls. Gene expression analyses of skin biopsies were performed using NanoString technology from patients with BP (n = 17), irBP (n = 19), and healthy skin (n = 24) after the patients had consented to participate in this study, and differentially expressed genes (DEGs) were determined using Rosalind software. Results: Compared to healthy skin, BP showed 167 DEGs, and irBP revealed 99 DEGs. Some of the DEGs from irBP and BP vs. healthy skin overlapped. Specifically, IL-4- and IL-13-associated genes were upregulated in both irBP and BP compared to healthy skin. Interestingly, expression profiles of BP vs. irBP also showed 13 DEGs. Conclusions: These findings suggest a possibility for therapeutic efficacy of IL-4 and IL-13 inhibitors in the treatment of irBP.

关键词： Periodontitis   Interleukin-17   Fenugreek   Phytochemicals   Molecular docking   In silico  

摘要： Periodontitis is a chronic inflammatory disease of the oral cavity, which is characterized by the excessive immune response to microbial stimuli and poses significant systemic health risks. The pro-inflammatory cytokine Interleukin-17 (IL-17) plays a pivotal role in the progression of periodontitis, by promoting immune cell recruitment and activating tissue-destructive molecules. This study explores the potential of fenugreek-derived phytochemicals-specifically isovitexin, rhaponticin, galactomannan, and quercetin-as inhibitors of IL-17 and its receptor, IL-17R, to mitigate inflammatory pathways in periodontitis. Using an in silico approach, molecular docking, molecular dynamics (MD) simulations, and binding affinity analysis were employed to assess the interaction of these compounds with the IL-17-IL-17R complex. Results indicate that isovitexin (IVI) and rhaponticin (RHA) exhibit strong binding affinities, with stable protein-ligand interactions throughout the simulations, supporting their potential as therapeutic agents. The study's findings contribute to the growing evidence of the anti-inflammatory benefits of fenugreek phytochemicals, suggesting that they may be utilized to develop novel therapeutic strategies for managing chronic inflammatory conditions such as periodontitis.