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关键词： Psoriasis   IL-36R   Neutrophil extracellular traps   NETosis  

摘要： NETosis happens when neutrophils are activated and neutrophil extracellular traps (NETs) are formed synchronously, which is a hallmark of psoriasis. However, the specific trigger that drives NET formation and the distinct contents and interaction with interleukin-36 receptor (IL-36R) of NETs remain to be further elucidated. This work identified NET formation driven by toll-like receptor (TLR) 3 ligand (especially polyinosinicpolycytidylic acid (Poly(I:C)) were enhanced by purinergic receptor P2X ligand-gated ion channel 7 receptor (P2X7R) ligands (especially adenosine 5 '-triphosphate '-triphosphate (ATP)). NET formation was accompanied by the secretion of inflammatory cytokines and characterized by IL-1 beta decoration. NET formation blockade decreased expressions of inflammatory cytokines and chemokines, which consequently improved inflammatory responses. Additionally, imiquimod (IMQ)-induced psoriasiform symptoms including neutrophilic infiltration tended to be time-sensitive. Mouse primary keratinocytes and mice deficient in Il1rl2, which encodes IL-36R, mitigated inflammatory responses and NET formation, thereby delaying the pathophysiology of psoriasis. Together, the findings provided the therapeutic potential for IL-36 targeting NET inhibitors in psoriasis treatment.

关键词： bergapten   NF-kappa B signaling pathways   phosphorylation   psoriasis  

摘要： Bergapten (BP) is a plant-derived furocoumarin that has a wide range of pharmacological effects. BP serves as a candidate amplifier in phototherapy against skin inflammation, such as psoriasis and atopic dermatitis. However, the anti-inflammatory role of BP remains elusive. We utilized IL-17A-stimulated keratinocyte line and imiquimod-challenged BALB/c mice to imitate psoriasis-like inflammation. Inflammatory phenotypes were determined by expressions of inflammatory genes and cytokines, histopathological changes and activities of nuclear factor-kappa B (NF-kappa B) pathway. An RNA-seq analysis of rodent skin was performed to explore possible mechanism lying behind. SiRNAs and antagonist (TMS) against cytochrome P450 family 1 subfamily B member 1 (CYP1B1) were subsequently used to determine the role of CYP1B1 in psoriasis pathogenesis in vitro and in vivo. Overexpression of CYP1B1 with lentivirus further validate therapeutic effect of BP. BP significantly suppressed activation of the NF-kappa B pathway by inhibiting p65 phosphorylation and improved the inflammatory phenotype both in vitro and in vivo. We revealed the key role of CYP1B1 in regulating the activation of the NF-kappa B signaling pathway. Knock-down with siRNAs significantly reduce the expression of inflammatory genes and cytokines. An intraperitoneal injection of TMS partially remediated IMQ-induced inflammation, mainly in terms of skin thickness. Overexpression of Cyp1b1 led to increased expression of the CYP1B1 protein and rescued the therapeutic effect of BP in vitro. This study revealed that BP suppressed expression of Cyp1b1 in keratinocytes and inhibited the activation of NF-kappa B signaling pathway by blocking the phosphorylation of p65.

关键词： TSLP   IL-4 receptor alpha   Humanized TSLP monoclonal antibody   Asthma   Combination therapy  

摘要： Background: Severe asthma is a complex and chronic respiratory disease, and current conventional treatments are not effective in controlling the patients' condition. Thymic stromal lymphopoietin (TSLP) is a key regulatory factor in the initiation and maintenance of asthma. Thus, blocking TSLP during allergic inflammation emerges as a promising therapeutic approach;however, novel anti-TSLP therapies remain to be developed. Furthermore, the importance of other signaling molecules, such as IL-4 and IL-13, should be considered. Moreover, to the best of our knowledge, the inhibitory effect of binding upstream and downstream signaling molecules has not been assessed. Purpose: This study aimed to develop a novel, humanized anti-TSLP antibody and explore the enhancement in its efficacy when combined with anti-IL-4R antibodies to treat asthma. Results: QX008N, derived from a rabbit antibody platform, exhibits a high affinity for TSLP and superior efficacy in blocking TSLP-induced signaling pathways and inflammation in vitro compared with Tezepelumab. In a cynomolgus monkey asthma model, QX008N ameliorated lung function and reduced the levels of eosinophils and IgE. Moreover, the coadministration of QX008N with anti-IL-4R antibodies enhanced the inhibition of inflammatory mediator production triggered via costimulation in vitro. In mouse asthma models, the simultaneous blockade of TSLP and IL-4R using anti-TL4R and anti-TSLP surrogates surpassed the efficacy of monotherapy. To the best of our knowledge, the therapeutic effect of a combination of anti-TSLP and IL-4R antibodies in an asthma model has not yet been reported. Conclusion: These results furnish comprehensive preclinical evidence for QX008N as an innovative anti-TSLP therapeutic agent and provide a preliminary rationale for the development of combination therapies that simultaneously target the TSLP and IL-4R signaling pathways.

关键词： PEDV   S1蛋白   单克隆抗体   抗原表位  

摘要： 为研制针对PEDV S1蛋白单克隆抗体，为下一步应用研究奠定材料基础，本研究构建了表达三聚化形态的S1蛋白的重组质粒，利用HEK 293F细胞表达并纯化获得S1蛋白，经免疫小鼠后，制备并筛选分泌S1特异性单克隆抗体的杂交瘤B细胞，制备腹水后利用间接ELISA法测定了腹水效价及单抗的亚型；通过SDS-PAGE、Western-blot、间接免疫荧光试验分析制备抗体与病毒S蛋白的特异性结合能力。结果显示，成功表达和纯化三聚化PEDV重组S1蛋白，通过免疫制备筛选了15株特异性识别病毒S蛋白的单克隆抗体。15株单克隆抗体的重链为IgG1型11株，IgG2b型3株，32#抗体未检测到分型结果。除25#抗体未检测到轻链结构外，其余抗体轻链均为kappa链。Western-blot和间接免疫荧光分析表明制备抗体与病毒S蛋白特异性反应。功能域结合试验表明抗体20#、21#、31#与S1 D0区域反应，抗体9#、16#、25#与S1 NTD反应，抗体15#、27#、37#与S1 SD2反应，所制备抗体均无病毒中和活性。结果表明，本研究成功制备了多株可特异性识别PEDV S1蛋白的单克隆抗体，为病毒感染机制研究及免疫学诊断研究提供了重要的生物材料。

摘要： Tumors escape immune detection and elimination through a variety of mechanisms. Here, we used prostate cancer as a model to examine how androgen-dependent tumors undergo immune evasion through downregulation of the major histocompatibility complex class I (MHCI). We report that response to immunotherapy in late-stage prostate cancer is associated with elevated MHC expression. To uncover the mechanism, we performed a whole genome CRISPRi screen and identified AR as a repressor of the MHCI pathway. Syngeneic mouse models of aggressive prostate cancer deficient in AR also demonstrated increased tumor immunogenicity and promoted T cell mediated tumor-control. Notably, the increase in MHCI expression upon androgen receptor blockade is transient and correlates with resistance to AR inhibition. Mechanistic studies identified androgen response elements upstream of MHCI transcription start sites which increased MHCI expression when deleted. Together, this body of work highlights another mechanism by which hormones can promote immune escape.

关键词： Editor: Noriyasu Hirasawa   MNSF beta   TNF alpha   Stress granules   Macrophage   Recurrent miscarriage  

摘要： Abnormally elevated tumor necrosis factor-alpha (TNF alpha) levels at the maternal-fetal interface can lead to adverse pregnancy outcomes, including recurrent miscarriage (RM), but the mechanism underlying upregulated TNF alpha expression is not fully understood. We previously reported that the interaction between monoclonal nonspecific suppressor factor-beta (MNSF beta) and RC3H1 upregulates TNF alpha expression, but the precise mechanisms are unknown. In this study, we found that MNSF beta stimulated the LPS-induced TNF alpha expression by inactivating the promoting effect of RC3H1 on TNF alpha mRNA degradation rather than directly inhibiting the expression of RC3H1 in THP1M phi s. Mechanistically, the 81-326 aa region of the RC3H1 protein binds to the 101-133 aa region of the MNSF beta protein, and MNSF beta facilitated stress granules (SGs) formation and the translocation of RC3H1 to SGs by interacting with RC3H1 and fragile X mental retardation 1 (FMR1) in response to LPS-induced stress. The SGslocalization of RC3H1 reduced its inhibitory effect on TNF alpha expression in LPS-treated THP1-M phi s. The designed HEPN2 peptide effectively reduced the LPS-induced expression of TNF alpha in THP1-M phi s by interfering with the MNSF beta-RC3H1 interaction. Treatment with the HEPN2 peptide significantly improved adverse pregnancy outcomes, including early pregnancy loss (EPL) and lower fetal weight (LFW), which are induced by LPS in mice. These data indicated that MNSF beta promoted TNF alpha expression at least partially by increasing the localization of RC3H1 to SGs under inflammatory stimulation and that the HEPN2 peptide improved the adverse pregnancy outcomes induced by LPS in mice, suggesting that MNSF beta is a potential pharmacological target for adverse pregnancy outcomes caused by abnormally increased inflammation at early pregnancy.

摘要： Lymphatic vessels play a crucial role in activating anti-tumor immune surveillance but also contribute to metastasis and systemic tumor progression. Whether distinct lymphatic phenotypes exist that govern the switch between immunity and metastasis remains unclear. Here we reveal that cytotoxic immunity normalizes lymphatic function and uncouples immune and metastatic potential. We find that in mice and humans, intratumoral lymphatic vessel density negatively correlates with productive cytotoxic immune responses and identify IFNγ as an intrinsic inhibitor of lymphangiogenesis. Specific deletion of the in lymphatic endothelial cells (LECs) greatly expanded the intratumoral lymphatic network and drove the emergence of a tip-like endothelial state, promoting lymph node metastasis but not dendritic cell migration. IFNγ inhibits oxidative phosphorylation, which is required for proliferation and acquisition of the pathologic transcriptional state. Our data indicate that IFNγ induces a phenotypic switch in tumor-associated lymphatic vessels to reinforce canonical immune surveillance and block metastasis.

关键词： Rheumatoid Arthritis   Triptolide   Curcumin   IL-17/NF-kappa B Signaling Pathway   Cell proliferation   Apoptosis  

摘要： Rheumatoid arthritis (RA) is a chronic, progressive, systemic autoimmune disease. While triptolide (TPL) and curcumin (CUR) are known to have multiple beneficial effects on RA, the combined effect of TPL and CUR remains unexplored. This study aimed to investigate their synergistic effect on cell proliferation and apoptosis via the IL-17/NF-kappa B signaling pathway. The collagen-induced arthritis (CIA) rat model was established, showing severe joint and synovial damage compared to normal rats. Treatment with TPL and CUR reduced the severity of RA in the CIA rat model and alleviated serum inflammatory cytokines, such as rheumatoid factor, IL-17, TNF-alpha, IL-1 beta, and IL-6. The elevated levels of IL-17 and NF-kappa B in CIA rats were also inhibited, and the resistant apoptosis was aggravated by TPL and CUR. In vitro, the improvement of cell proliferation and induction of apoptosis were observed in LPS-stimulated MH7A cells treated with TPL and CUR, associated with the inhibition of the IL-17/NF-kappa B signaling pathway. Taken together, a synergistic effect of TPL and CUR on RA may involve relieving symptoms, improving excessive proliferation, inducing apoptosis resistance, and inhibiting the IL-17/NF-kappa B signaling pathway.

关键词： 扶正辟邪颗粒   新冠病毒S蛋白   系统性炎症   调节免疫功能   细胞因子  

摘要： 目的 评价扶正辟邪方调节免疫功能的作用，为其临床应用提供科学的参考依据。方法 利用多次注射新冠病毒S蛋白后再联合LPS诱导小鼠产生多系统炎症、腹腔注射环磷酰胺建立免疫功能低下小鼠模型考察扶正辟邪方对机体免疫功能的调节作用。结果 扶正辟邪方显著升高IL-6、IL-10、MCP-1、MIP-1α和RANTES水平，减轻系统性炎症反应小鼠肺间质炎细胞浸润以及非间质增宽程度，改善小鼠脾脏白髓增生、副皮质区萎缩、红髓萎缩和星空细胞增生，抑制小鼠体温降低；扶正辟邪方可以显著增加小鼠巨噬细胞的吞噬能力。结论 扶正辟邪方可显著改善S蛋白及LPS诱导系统性炎症，可有效预防新冠病毒感染，其作用机理可能与其具有调节细胞因子风暴、调节免疫功能等作用有关。

关键词： Interleukin-8   CXCR 1/2   E-cadherin   ZO-1   Lens epithelial cells   PCO   RhoA  

摘要： Background: Posterior capsular opacification is a major complication following cataract surgery, marked by proliferation, migration, epithelial-mesenchymal transition, and fibrosis of residual epithelial cells. Various inflammatory cytokines are upregulated and contribute to the development of posterior capsular opacification. The effect of interleukin-8 on residual epithelial cells has not been fully determined. Methods: Aqueous humor and anterior capsules samples were collected from cataract surgery. Capsular bags from rats and pigs were cultured in DMEM media. Protein and mRNA expressions were measured using immunoblot and qPCR. Cell migration was assessed using the transwell assay. Results: Interleukin-8 is an early inflammatory factor secreted by residual lens epithelial cells. Migration of lens epithelial cells in aqueous humor positively correlates with interleukin-8 levels, and this effect is inhibited by the receptors of interleukin-8 CXCR1/2 blocker Reparaxin. The expression of tight-junction protein ZO-1 and celladhesion protein E-cadherin were down-regulated by administrating interleukin-8, and cell migration of both SRA01/04 cell line in vitro and capsular residual epithelial cells ex vivo were up-regulated via activating RhoA expression and RhoA/GTPase activity. The loss-of- function studies demonstrate that interleukin-8 binding to its receptor CXCR1/2 activates NF-kappa B/p65, which then turns on the RhoA's expression and RhoA/GTPase activity, and RhoA-modulated the downexpression of E-cadherin and ZO-1 and the increase of cell migration. Conclusions: The upregulation in interleukin-8 occurs early in posterior capsular opacification and contributes to down-regulating tight-junctions among epithelial cells and elevates cell migration via the CXCR1/2-NF-kappa B-RhoA signaling pathway. These demonstrated that interleukin-8 could be a potential target for preventing posterior capsular opacification.