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关键词： NK cell   CAR NK cell   Cord blood   Progenitor cell   IL-15   Adoptive cell therapy  

摘要： Cord blood (CB)-derived chimeric antigen receptor (CAR) natural killer (NK) cells have demonstrated significant antitumor efficacy. We recently reported that CB-derived CAR NK cells predominantly originate from CD7+CD56-CD34-HLA-DR-Lin- NK cell precursors in CB. Here, we demonstrate that stimulating the interleukin (IL)-15 receptor on these NK precursors enhances the production of CAR NK cells from CB cells. In CB CD56-CD34-HLA-DR-Lin- cells, the IL-15 receptor was exclusively expressed on CD7+ NK cell precursors. Using K562 feeder cells that express not only 4-1BB ligand and membrane-bound (mb) IL-21 but also mbIL-15 significantly increased the production of mature NK cells from the purified NK cell precursors or T cell-depleted CB cells. The in vitro and in vivo antitumor effects of CAR NK cells generated using K562 feeder cells that express mbIL-15 were comparable to those of CAR NK cells produced using K562 feeder cells that do not express mbIL-15. These results suggest that K562 feeder cells expressing 4-1BBL, mbIL-21, and mbIL-15 can increase the production of CAR NK cells from CB cells while maintaining their cytotoxic potential. This method could also be useful for expanding NK cells from CB for any type of adoptive NK cell therapy with or without CAR transduction.

摘要： Background:Thyroid nodules are common in clinical practice, with malignancy detected in about 5% of cases. Current risk-stratification approaches, which rely on sonographic features and cytopathological assessment, have notable limitations, especially for indeterminate nodules categorized as Bethesda III. Aims:To address these diagnostic challenges, this study evaluated the utility of preoperative serum interleukin (IL)-17A and calprotectin levels as potential biomarkers for distinguishing malignant from benign Bethesda III thyroid nodules. Study Design:This single-center prospective methodological study. Methods:In this prospective study, 76 patients with Bethesda III nodules scheduled for thyroidectomy were enrolled. Based on histopathological findings, nodules were classified as benign (n = 41) or malignant (n = 35). Comprehensive patient information was collected, including demographics, medical history, and detailed clinical parameters related to nodule characteristics and laboratory results. Circulating biomarkers measured included thyroid-stimulating hormone, free T3, free T4, thyroglobulin (TG), anti-TG, anti-thyroid peroxidase, calcitonin, IL-17A, and calprotectin. Results:Patients with malignant nodules (n = 35) exhibited significantly higher preoperative IL-17A and calprotectin levels compared with those with benign nodules (n = 41) ( < 0.001 and = 0.038, respectively). receiver operating characteristic analysis demonstrated promising diagnostic performance for IL-17A [area under the curve (AUC) = 0.733] and calprotectin (AUC = 0.639). Conclusion:IL-17A and calprotectin emerge as promising biomarkers for refining Bethesda III nodule stratification. Incorporating these inflammatory markers into existing diagnostic protocols may substantially reduce unnecessary surgical interventions, thereby alleviating patient anxiety, surgical risks, and healthcare costs.

摘要： Background We sought to determine the prevalence, clinical impact, neutralizing capacity, and cross-reactivity of pre-existing antibodies to infliximab and adalimumab in antitumor necrosis factor (TNF) treatment-naive patients with active luminal crohn disease. Methods The prevalence of antibodies to infliximab and adalimumab at entry to the "Personalised Anti-TNF Therapy in Crohn's Disease Study" (PANTS) were measured using the drug-tolerant IDKmonitor Total antidrug antibody ELISAs. Neutralizing capacity in positive cases was determined using iLite (TM) cell-based assays. Results Pre-existing antibodies to infliximab were more common (6.3%, 96/1525 vs 2.4%, 36/1525, P < 0.01) and detectable at higher concentrations [median (IQR) 23.9 (14.2-55.2) AU/mL vs 7.2 (6.3-10.4) AU/mL, P < 0.0001] to infliximab than adalimumab. A few patients [1.3% (95% CI 0.6-1.8) 20/1525] had antidrug antibody reactivity to both drugs. None of the detected antidrug antibodies had demonstrable anti-TNF neutralizing capacity. No associations were seen between pre-existing antibody positivity, adverse drug reactions, drug levels, or response status at weeks 14 or 54 or subsequent immunogenicity. Cross-reactivity with rheumatoid factor and antimouse antibodies was detected (>10 IU/mL) in 6.6% (95% CI 3.2-13.0) (7/106) and 17.0% (95% CI 10.5-25.2) (19/112) of patients with pre-existing antibodies. Conclusions Pre-existing antidrug antibodies are common but do not neutralize anti-TNF drug activity in vitro, promote drug clearance, or influence clinical response to anti-TNF drugs. Further work is required to understand this cross-reactivity and to determine the exact nature of the antibodies detected in drug-naive individuals.

关键词： 非洲猪瘟病毒   多表位蛋白   触发因子   免疫原性  

摘要： 旨在制备非洲猪瘟病毒（ASFV）多表位抗原并评价其在疫苗中应用。本研究设计了分子伴侣（TF16）与ASFV多表位抗原原核重组质粒，可溶性表达非洲猪瘟病毒多表位重组蛋白（ASFV-pBT），纯化鉴定后免疫小鼠并进行免疫原性评价，为非洲猪瘟亚单位疫苗研究提供试验基础。结果表明，ASFV-pBT可在大肠杆菌中可溶性表达，并被ASFV抗体阳性血清特异性识别。免疫小鼠血清中抗体效价可达1:32 000，且ASFV-pBT可有效激活小鼠T淋巴细胞的增殖，CD4+、CD8+T淋巴细胞较阴性对照显著提升（P<0.001），IFN-γ和IL-4细胞因子的表达也显著高于阴性对照组（P<0.001）。通过体内、体外双重验证表明ASFV-pBT重组蛋白展现出良好的免疫原性，能够有效诱导机体产生显著的体液免疫和细胞免疫，为ASFV表位疫苗的开发提供了参考。

关键词： Sterols   LXR   IL-17   Acute inflammation   Alleviation  

摘要： Inflammation plays a crucial role in defending the host against pathogens and other harmful stimuli. However, uncontrolled acute inflammation has been implicated in a multitude of diseases, including those that affect aquatic animals. While the processes that initiate the inflammatory response are relatively well understood, the roles of lipids, their derivatives, and the mechanisms by which they contribute to alleviating acute inflammation remain less well characterized. In the present study, lipidomic analyses were performed to identify changes in lipid metabolism in the coelomocytes of the sea cucumber Apostichopus japonicus after infection with Vibrio splendidus. The omics data indicated significant increases in specific sterols, including desmosterol, 25-hydroxycholesterol, and 24,25-epoxycholesterol, during the late phase of the inflammatory response. The accumulation of these sterols was found to be responsible for the suppression of A. japonicus interleukin-17 (AjIL-17)-mediated inflammatory response. A subsequent biolayer interferometry assay revealed that A. japonicus liver X receptor (AjLXR) served as the downstream receptor of these sterols. The knockdown of AjLXR reversed the desmosterol-dependent suppression of AjIL-17 expression. Conversely, the activation of AjLXR by the synthetic agonist GW3965 inhibited the expression of AjIL-17 induced by infection with V. splendidus or stimulation with LPS. Moreover, AjLXR repressed AjIL-17 transcription by directly binding to the AjIL-17 promoter in a SUMOylation-independent manner. In conclusion, our findings highlight the crucial role of the sterols-LXR axis in mitigating IL-17-mediated acute inflammation, suggesting that targeting this axis could be a promising strategy for preventing and treating chronic inflammation-associated diseases.

摘要： Natural killer (NK) cell function is impaired in patients with chronic hepatitis B (CHB) infection;however, the underlying mechanisms are not fully understood. Here, we collected the blood samples from healthy donors (HDs) and patients with CHB, and then analyzed the characteristics of NK cells by RNA-seq analysis, flow cytometry, Seahorse assay. HBV-carrier mice were used to confirm the findings in vivo. We found that the dysfunction of NK cells in peripheral blood of patients with CHB was associated with the disturbance of glycolysis. Further investigation showed chronic HBV infection impaired the activation of mammalian target of rapamycin (mTOR) in NK cells, resulting in decreased expression of molecules involved in glycolysis, including HIF-1 alpha and GLUT1. Mechanistically, we found that HBsAg suppressed IL-15-triggered mTOR activity by competitively binding to the IL-15 receptor beta (IL-15R beta, CD122) on NK cells, leading to the decreased expression of HIF-1 alpha and its downstream genes. Significantly, HBsAg neutralizing antibody intravenous injection or mTOR agonist MHY1485 intraperitoneal injection restored the IL-15/mTOR signaling in NK cells of HBV-carrier mice, resulting in NK cell activation and HBV clearance. Further, transferring MHY1485-pretreated NK cells isolated from HBV-carrier mice displayed augmented anti-HBV effects in recipient HBV-carrier mice. These findings reveal a new mechanism by which chronic HBV infection induces NK cell dysfunction, and highlight the potential of mTOR activation and HBsAg clearance as therapeutic strategies for CHB treatment via recovering NK cell immune functions.

摘要： Natural killer (NK) cells play a critical role in anti-cancer immunity through their direct cytotoxicity and production of cytokines, such as Flt3L. NK cell production of Flt3L controls conventional type I dendritic cell (cDC1) abundance in the tumor and promotes protective immune responses. Here, we show that NK cell production of Flt3l in the tumor is regulated by activation, and that activation by IL-2 and IL-15 uniquely induced Flt3L expression in NK cells. In melanoma, IL-2 signaling in NK cells led to increased Flt3L production, which boosted cDC1 abundance in the tumor and improved anti-PD-1 immunotherapy response. Further, NK cell subsets differentially regulated Flt3L in the tumor, with CD11b-CD27+ NK cells in mouse tumors enriched for IL-2-family signaling and upregulated Flt3l upon activation. Consistently, human CD56brightCD16- NK cells more strongly correlated with cDC1 and FLT3LG expression than other NK cell subsets across multiple human melanoma datasets and cancer indications. This mechanistic study of NK cell regulation of FLT3LG and control of the NK cell-cDC1 axis provides insights and strategies for the development of more effective cancer immunotherapies.