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关键词： 赤羽病毒   Gc蛋白   原核表达   单克隆抗体  

摘要： 为了进一步研究赤羽病毒的感染机制和Gc蛋白的功能，利用原核表达系统成功表达赤羽病毒的Gc蛋白。用纯化后的重组蛋白免疫BALB/c小鼠，经过4次免疫、间接ELISA筛选和细胞融合，获得1株能够分泌抗Gc蛋白单克隆抗体的细胞株，命名为2D3，其抗体亚型为IgG1/Kappa型。IFA、Western-blot和间接ELISA结果表明，单克隆抗体2D3具有高特异性和亲和性，能特异性识别赤羽病毒Gc蛋白。本研究成功制备了抗赤羽病毒Gc蛋白的单克隆抗体2D3，为赤羽病毒检测方法的建立及病毒致病机制研究提供了可靠的工具。

摘要： Chronic wound healing, particularly in diabetic wounds, remains a global challenge. The delayed or even stagnant healing process imposes a substantial burden on both society and individuals. A critical challenge lies in effectively reducing inflammatory levels and promoting vascular remodeling. As an adipokine, C1QTNF9 (CTRP9) has been demonstrated to suppress inflammatory responses under various pathological conditions. In diabetic wound healing, CTRP9 facilitates the proliferation, differentiation, and migration of keratinocytes, yet its specific regulatory mechanisms on local inflammation remain unclear. Transcriptomic analysis of diabetic wounds revealed significantly reduced CTRP9 transcription levels in non-healing wounds. Animal experiments showed lower CTRP9 expression in the skin tissues of diabetic mice compared to normal mice, suggesting a close association between CTRP9 and wound healing. Subsequent local supplementation of CTRP9 in diabetic mouse wounds significantly accelerated healing to near-normal levels. Notably, SLC7A11 (also known as xCT)-a key protein linked to impaired efferocytosis in dendritic cells at diabetic wounds-was downregulated by CTRP9, while CTRP9 enhanced efferocytic efficiency. ATF3, a transcriptional repressor of SLC7A11 involved in ferroptosis regulation, has not been previously explored in dendritic cell efferocytosis. This study found that ATF3 knockdown upregulated SLC7A11 expression and attenuated CTRP9-mediated efferocytosis enhancement, indicating ATF3 as a critical pathway through which CTRP9 regulates SLC7A11. Furthermore, CTRP9 enhances glycolysis to produce lactate, thereby providing energy support for dendritic cell efferocytosis. In summary, this study demonstrates that CTRP9, as an anti-inflammatory factor, promotes diabetic wound healing by enhancing dendritic cell efferocytosis and reducing local inflammation, offering a promising therapeutic strategy.

关键词： 丹参多糖   Mn   PLGA   佐剂   细胞免疫   体液免疫  

摘要： 本研究利用PLGA包封了免疫刺激效果强和生物安全性良好的丹参多糖和Mn2+，评价了该体系作为灭活PCV2佐剂的免疫增强效果。首先我们利用复乳溶剂挥发法制备了共载丹参多糖和Mn2+的PLGA纳米佐剂递送系统（SM-PLGA），将SM-PLGA与PCV2抗原混合，对小鼠左右大腿皮下分别注射100 μL进行初免，同时设立空白对照组、PCV2组、Alhydrogel （Algel）-PCV2组、丹参多糖-PLGA组、SM组、丹参多糖组和PLGA组，免疫后的第14天对小鼠进行加强免疫。初免后的第5天通过流式细胞术评估腘淋巴结和腹股沟淋巴结中DCs的活化情况。在初免后的第21、28、35、42天收集小鼠血清，对PCV2特异性抗体IgG、IgG1、IgG2a进行动态监测，评价佐剂对PCV2特异性体液免疫反应的增强效果。在初免后的第21、28天采集小鼠脾脏淋巴细胞，分别对CD4+T细胞、CD8+T细胞、细胞毒性T细胞、记忆性T细胞和生发中心B细胞活化情况进行测定，并对第28天细胞上清中INF-β、INF-γ、IL-6进行测定。最后，在初免后的第35天采集小鼠淋巴结，测定生发中心B细胞的活化情况。通过上述不同时段的测定结果，评价SM-PLGA对机体免疫反应的增强效果。由结果可知SM-PLGA在初免后的第5天可以极显著促进淋巴结DCs的活化，在初免后第21～42天可以持续刺激PCV2特异性抗体IgG及其亚型的分泌。在初免后第21天SM-PLGA可显著促进脾脏CD4+T细胞、CD8+T细胞和CD8+T细胞CTL应答，并在初免后第28和35天仍对记忆性T细胞和生发中心B细胞有较强的刺激作用，同时极显著促进相关细胞因子IFN-β、IFN-γ、IL-6的分泌。结果表明SM-PLGA作为灭活PCV2的佐剂，可以诱导强效的Th1/Th2免疫应答，为PCV2的预防提供参考。

关键词： Disease Modification   Early Intervention   IL-17 Inhibitor   Ixekizumab   Psoriasis   Secukinumab  

摘要： BACKGROUND:Although IL-17 inhibitors like Secukinumab and Ixekizumab have shown significant efficacy in psoriasis, the impact of early intervention with biologics to modify the disease course and achieve long-term remission remains unclear. OBJECTIVES:To examine the potential of early intervention with IL-17 inhibitors for disease modification in psoriasis. METHODS:We conducted a multicenter retrospective cohort study on moderate-to-severe plaque psoriasis patients who received at least 4 weeks of treatment with Secukinumab or Ixekizumab between April 2019 and April 2023, taking the relapse rate one year after cessation of treatment as the primary endpoint. RESULTS:Among 400 patients who discontinued treatment after achieving PASI90, the median relapse time was 3.29 months(approximately 14 weeks). Of 141 patients who discontinued treatment after achieving PASI90 for over a year, 24 (88.89%) in the ultra-short disease duration(USDD, psoriasis duration ≤ 1 year) group and 33 (82.5%) in the short disease duration(SDD, psoriasis duration ≤ 2 year) group achieved one year of drug-free remission. LIMITATIONS:The potential impact of early intervention on comorbidity development was not addressed in this study. CONCLUSION:Early intervention with IL-17 inhibitors leads to faster responses and may promote disease modification in psoriasis.

关键词： Epitope prediction   Epitope presentation   Helper T cells   SARS-CoV-2   Spike protein  

摘要： Early in the COVID-19 pandemic, multiple studies used prediction methods to identify candidate peptides to be included in prospective vaccines. While subsequent studies identified epitopes from convalescent and vaccinated subjects, few studies have compared the predicted to identified epitopes. Here we used three methods to predict SARS-CoV-2 spike protein helper T cell epitopes and compared the results to experimentally determined peptide binding as well as published epitopes. The correspondence between the predicted and experimental binding results and published epitopes depended more on the HLA being investigated than the prediction method used. Lastly, these observations were used to predict peptides which bind to the most HLAs. These peptides were previously identified and predicted to maintain HLA binding in the current variants of interest. This study highlights which prediction methods and conditions lead to the most reliable prediction results which would be of great interest for improving the design of future vaccines.

摘要： Cytokines play a substantial role in the pathophysiology of infectious diseases. Consequently, genetic polymorphisms in cytokine genes can have profound effects on the susceptibility to or protection from infections and also influence clinical manifestations, disease severity, as well as outcome. This narrative review summarizes the recent findings on the role of genetic polymorphisms of IL-18, a recently discovered cytokine, on certain infectious diseases of public health relevance. The findings of the review suggest that IL-18 polymorphisms can have a significant impact on multiple infectious diseases, including malaria, leishmaniasis, HIV/AIDS, HBV, tuberculosis and H. pylori. This highlights the need to understand specific polymorphisms to decipher their protective or predisposing effects. For instance, the presence of G and T alleles at position − 656 is suggested to have a probable protective effect against Visceral Leishmaniasis. Haplotypes −137 G/−607 C (GC) and −137 C/−607 C (CC) in the promoter region are associated with increased susceptibility to severe malarial anaemia. SNPs rs544354 and rs574429 (GC) are linked to a higher risk of P. falciparum infection and increased parasitemia. The −137 C/C genotype and −137 G allele (with G/G genotype) are associated with faster progression to AIDS. In contrast, the IL-18 genotype does not seem to affect HIV susceptibility. IL-18 polymorphisms influence the pathophysiology in infections like HBV, H. pylori, and TB. However, the conflicting results in studies conducted in different populations (e.g., genetic polymorphism, studies conducted in India and China in HBV infection) necessitate further research with larger sample sizes to resolve inconsistencies in the findings.

关键词： Astragalus injection   Melanoma   Anti-PD-1   MHC-II   STAT1   CIITA  

摘要： Ethnopharmacological relevance: Astragalus injection, a solution derived from Astragalus mongholicus Bunge, has been used in cancer patients for its immune-boosting properties. The effects of Astragalus injection combined with immune checkpoint inhibitors (ICIs) in cancer remain unsuspected. Aim of the study: This study aims to investigate the efficacy of combining Astragalus injection with anti-PD-1 and to elucidate the potential mechanisms. Materials and methods: A melanoma-bearing mouse model was established to observe the effects of Astragalus injection combined with anti-PD-1 treatment. The immune cell infiltration in tumor tissues was evaluated by flow cytometry. Subsequently, network pharmacology and RNA sequencing were conducted to anticipate latent mechanisms of Astragalus in melanoma. Finally, these predictions were validated by flow cytometry, qPCR, Western blotting, and ELISA. Results: Astragalus injection significantly antagonized the efficacy of anti-PD-1 in melanoma, resulting in a notable reduction of immune cell infiltration. The network pharmacology and RNA sequencing analysis revealed critical signaling routes, encompassing T-cell receptor activation, immune antigen presentation, and the JAK/ STAT pathway. Subsequent data suggested that Astragalus injection could down-regulate the expressions of MHCII in dendritic cells and B16-OVA cells and restrict the activation of dendritic cells. Moreover, the expressions of CIITA and phosphorylated STAT1 were prominently inhibited by Astragalus injection, whereas the overexpression of STAT1 partially reversed the Astragalus injection-induced decrease of MHC-II. Conclusions: Astragalus antagonized the therapeutic effect of anti-PD-1 in melanoma. These effects were partially through inhibiting JAK/STAT signaling and down-regulating MHC-II expression.

关键词： RAB3IL1   SLC7A11   ferroptosis   male infertility   spermatogenesis microenvironment  

摘要： Ferroptosis, an iron-dependent form of regulated cell death characterized by phospholipid peroxidation, plays a pivotal role in various diseases. However, its involvement in male infertility, particularly in idiopathic non-obstructive azoospermia (iNOA), remains largely unexplored. iNOA accounts for 10-15% of male infertility cases, presenting a significant clinical challenge due to its unknown origins. This study investigated the potential link between ferroptosis and iNOA, revealing excessive activation of both ferroptosis and autophagy in the testes of iNOA patients, along with severe disruption of the blood-testis barrier (BTB). The BTB, formed by tight junctions between Sertoli cells, is essential for maintaining the spermatogenesis microenvironment. Using the ferroptosis inducer erastin, an antagonist of the cystine antiporter SLC7A11, we established a ferroptosis model in mouse Sertoli cells, demonstrating that ferroptosis activation led to cytoskeletal disarray and BTB disruption, accompanied by excessive autophagy activation. Notably, inhibition of autophagy using 3-methyladenine significantly rescued erastin-induced ferroptosis and restored BTB integrity, highlighting a crucial cross-talk between ferroptosis and autophagy in maintaining the spermatogenic microenvironment. Furthermore, through combined transcriptome analysis of iNOA testes and Sertoli cells, we identified RAB3IL1 as a novel regulator of ferroptosis. Mechanistically, Rab3il1 knockdown induced ferroptosis via translocation of SLC7A11, resulting in cytoskeletal defects and BTB damage. Our findings underscore the critical role of the ferroptosis-autophagy axis, regulated by RAB3IL1, in preserving the spermatogenic microenvironment, offering a potential therapeutic target for restoring spermatogenesis in iNOA patients.

关键词： DILI   DRESS   Lamotrigine   Phenytoin   Trimethoprim-Sulfamethoxazole  

摘要： INTRODUCTION:Drug reaction with eosinophilia and systemic symptoms (DRESS) can sometimes occur in patients with drug-induced liver injury (DILI). However, detailed studies of DRESS in patients with DILI from the US are lacking. We investigated the characteristics and HLA risks for DILI who also developed DRESS. METHODS:Patients with definite, highly likely, or probable DILI enrolled into US DILI Network studies between September 2004 and August 2023 were included. DRESS was defined based on modified RegiSCAR criteria. HLA alleles were compared between DILI-DRESS cases and two control groups (DILI with non-DRESS rash [n=244] and DILI without rash [n=1637). RESULTS:Of 2,121 participants with DILI during the study period, 128 participants had DRESS (6%). The most frequently implicated drugs causing DRESS were trimethoprim/sulfamethoxazole, lamotrigine, phenytoin, allopurinol, and vancomycin. Compared to 1993 DILI patients without DRESS, patients with DILI + DRESS were younger (mean age 42.3 yrs vs 50.6 years), more likely to be Black (26% vs 12%), had shorter latency (median 31 days vs 47 days), higher frequency of rash (100% vs 13%), eosinophilia (55% vs 13%), and fever (76% vs 16%) (P<0.001 for all). Compared to DILI without DRESS, DILI + DRESS had more severe liver injury (severe/fatal: 45% vs 21.5%, p<0.001) and higher overall (15.6% vs 6.3%, P<0.001) and liver-related (9% vs 2.3%, p < 0.001) mortality. HLA A*32:01, HLA B*53:01 and HLA B*58:01 were significantly enriched in DILI-DRESS cases, compared to control groups. CONCLUSIONS:Patients with DILI and DRESS are younger, more likely to be Black, had shorter time to DILI onset with more severe liver injury and higher overall and liver-related mortality. HLA A*32:01, HLA B*53:01 and HLA B*58:01 are risk factors for DILI-DRESS.