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关键词： MHC class I   antigen presentation   antigen processing   endoplasmic reticulum aminopeptidase   ovalbumin   peptide trimming  

摘要： Peptide presentation by major histocompatibility complex (MHC) class I molecules enables CD8+ T lymphocytes to monitor the intracellular proteome of tissue cells. CD8+ T cell priming and acquisition of effector functions is affected by cognate peptide-MHC-I complex density on the cell surface, which partly depends on the efficacy of intracellular proteolytic peptide generation. Peptide generation frequently requires final trimming by the human aminopeptidases ERAP1, ERAP2, and IRAP. All display genetic polymorphism associated with the risk of multiple autoimmune diseases but also some cancers. This finding has prompted interest in the development of small molecule inhibitors to enhance antitumor or conversely attenuate autoreactive T cell responses. However, efficient assays for assessment of inhibitor effects are wanting. We describe the development of an assay for quantitative assessment by flow cytometry of selective inhibitor effects on peptide trimming both in the endogenous MHC-I processing pathway and in cross-presentation. We use the assay to identify a selective ERAP2 inhibitor and show that inhibitor effects can be read out not only through assessing a specific peptide-MHC complexes but also by measuring cell surface levels of bulk MHC-I molecules. Next to its practical interest as tool for inhibitor testing, our assay highlights how ERAP1-dependent immunodominance of a single epitope processed with exceptional efficacy can have a massive effect on the immunopeptidomic identity of cells presented to CD8+ T cells. We propose that ERAP effects on the presentation of such rare and exceptionally immunodominant epitopes may underlie the epistatic genetic associations of ERAP polymorphism with HLA class I-linked autoimmune diseases.

关键词： FOXP3   IL-2   interleukin-2   regulatory T cells   Tregs  

摘要： MK-6194, an interleukin-2 mutein designed to selectively activate regulatory T cells (Tregs), was evaluated for safety, pharmacokinetics (PK), immunogenicity, and pharmacodynamics in healthy participants. In a single ascending dose trial (N = 56), participants received subcutaneous MK-6194 or placebo (3:1 ratio) across dose levels ranging from 1 to 10 mg. In a multiple ascending dose trial (N = 54), participants received subcutaneous MK-6194 or placebo (3:1 ratio) at dose levels ranging from 0.5 to 5 mg every 2 wk (total 3 doses) as well as 5 mg every 4 wk (total 2 doses). Baseline characteristics were comparable between trials, with participants mostly male with a mean age of 36 yr. There were no serious adverse events or dose-limiting toxicities. The most common adverse events were injection site erythema and eosinophil count elevations (with no indication of severe eosinophilia or eosinophilia-related organ damage). PK showed dose-proportionality and repeated doses of MK-6194 did not result in accumulation or time-dependent PK. Immunogenicity was low with no impact on PK or safety. Treg expansion as assessed by flow cytometry and Treg-specific demethylation region analysis was observed in a dose-dependent manner during both trials and expanded within about 8 d postdose up to about 5-fold and returned to baseline by 14 to 29 d postdose. Minimal impact was observed on other lymphocytes including total T lymphocyte and natural killer cell counts. These findings support the further development of MK-6194 as a potential treatment for autoimmune disorders. © The Author(s) 2025. Published by Oxford University Press on behalf of The American Association of Immunologists.

关键词： pediatric patients   therapeutic drug monitoring (TDM)   anti-TNF biological agents   enzyme-linked immunosorbent assay (ELISA)   fluorescence-based lateral flow immunoassay (AFIAS)   turnaround time (TAT)  

摘要： Background: Therapeutic drug monitoring (TDM) of infliximab (IFX) and adalimumab (ADL) mainly relies on the use of enzyme-linked immunosorbent assays (ELISA). More recently, rapid assays have been developed and validated to reduce turnaround time (TAT). Here, we compared IFX and ADL concentrations measured with both ELISA and a new fluorescence-based lateral flow immunoassay (AFIAS). Methods: In serum samples from pediatric patients, IFX and ADL drug levels, and total anti-IFX antibodies were measured using clinically validated ELISA kits (Immundiagnostik AG). Samples were further analyzed using a new rapid assay (AFIAS, Boditech Med Inc.) to measure drug levels and total anti-IFX antibodies. Results: Spearman's correlation coefficients (rho) were 0.98 [95% confidence interval (CI) 0.97 to 0.99] for IFX (p < 0.001) and 0.83 (95% CI 0.72 to 0.90) for ADL (p < 0.001). Calculated % bias was -14.09 (95% Limits of agreement, LoA, -52.83 to 24.66) for IFX and 15.79 (LoA -37.14 to 68.73) for ADL. For the evaluation of total anti-IFX antibodies, we did not collect sufficient data to establish a statistically significant correlation between AFIAS and ELISA. The inter-rater agreement showed a "substantial" and a "moderate" agreement for IFX and ADL, respectively. Conclusions: Our results show that the AFIAS assay has an accuracy and analytical performance comparable to that of the ELISA method used for TDM of IFX and ADL. Therefore, the introduction of this device into routine clinical practice could provide results more quickly and with similar accuracy as ELISA, allowing clinicians to rapidly formulate clinical decisions.

关键词： Lung cancer   Radiation bystander effect   Intestinal injury   Isoliquiritigenin   TNF-alpha  

摘要： To construct a model of intestinal injury induced by radiotherapy for lung cancer and to study the protective effect and mechanism of isoliquiritigenin. The lungs of mice were irradiated with 0, 2, 4, 6, 8 Gy X-rays to screen the optimal radiation dose. A mouse model of lung cancer was established, and the tumor was irradiated once. At 3, 7, and 10 days after irradiation, H&E was used to detect the pathological manifestations of colon tissue in mice, and WB was used to detect the expression level of tight junction protein in colon tissue, so as to screen the best time point and study its possible mechanism. Molecular docking was used to study the tightness of isoliquiritigenin binding to TNF-alpha. Isoliquiritigenin (40 mg/kg) was given on the next day after 4 Gy X-ray irradiation. The levels of TNF-alpha and apoptosis, intestinal mucosal barrier function, MUC2 protein expression, and colon stem cell proliferation were detected. 4Gy X-ray local irradiation induced obvious colon injury in mice, and the injury was obvious on the 7th day. Isoliquiritigenin significantly improved the general condition, colonic histopathological changes, intestinal stem cell proliferation, and colonic tight junction function of lung cancer-bearing mice after radiotherapy. Further studies have found that isoliquiritigenin can downregulate the activation of TNF-alpha/Caspase-3 signaling pathway by inhibiting the expression of pro-inflammatory factor TNF-alpha, alleviate the apoptosis of colonic epithelial cells, improve the upregulation of colonic tight junction function, regulate the expression of MUC2, and promote the proliferation of intestinal stem cells, which may be related to the stable binding of isoliquiritigenin to TNF-alpha. Radiotherapy-induced bystander effect of lung cancer may be related to the abnormal expression of TNF-alpha. Isoliquiritigenin may downregulate the expression of TNF-alpha by binding to TNF-alpha, inhibit the apoptosis of colon cells, promote the proliferati

关键词： 血型同种免疫   CD4   T细胞   B细胞   细胞免疫平衡   综述  

摘要： 慢性贫血患者（如地中海贫血）需长期依赖输注红细胞维持生命，但血型抗原的同种免疫反应常给患者的临床治疗与生存带来严重危害。输血相关同种免疫的调控机制尚不明确，如有些患者尽管长期输血，不会发生同种免疫反应；而有些患者会产生针对多种血型抗原的同种抗体，使输血治疗变得越来越困难。红细胞血型同种免疫需多种免疫细胞参与，包括抗原递呈细胞，不同的T细胞等，许多研究都在探索相关的调节作用，甚至干预措施。本文综述了细胞免疫与红细胞血型同种免疫的相关性，探讨二者之间的相互作用及其在免疫应答中的影响。

摘要： Gastrointestinal (GI) dysfunction emerges years before motor symptoms in Parkinson's disease (PD), implicating the enteric nervous system (ENS) in early disease progression. However, the mechanisms linking the PD hallmark protein, α-synuclein (α-syn), to ENS dysfunction - and whether these mechanisms are influenced by inflammation - remains elusive. Using iPSC-derived enteric neural lineages from patients with α-syn triplications, we reveal that TNF-α increases mitochondrial-α-syn interactions, disrupts the malate-aspartate shuttle, and forces a metabolic shift toward glutamine oxidation. These alterations drive mitochondrial dysfunction, characterizing metabolic impairment under cytokine stress. Interestingly, targeting glutamate metabolism with Chicago Sky Blue 6B restores mitochondrial function, reversing TNF-α-driven metabolic disruption. Our findings position the ENS as a central player in PD pathogenesis, establishing a direct link between cytokines, α-syn accumulation, metabolic stress and mitochondrial dysfunction. By uncovering a previously unrecognized metabolic vulnerability in the ENS, we highlight its potential as a therapeutic target for early PD intervention.

关键词： Enterovirus 71   IFN alpha   Antiviral activity   HMC3 cells   MiR-362-3p  

摘要： Enteroviruses, which belong to the Picornaviridae family, include species that infect humans and Interferon Alpha (IFN alpha) is commonly used against Enterovirus 71 (EV71) infections. This study investigated the role of IFN alpha-induced miR-362-3p expression in the defense against EV71EV71. Differential analysis identified up-regulated miRNAs following IFN alpha treatment. RD cells were used to assess EV71proliferation, while HMC3 cells were employed to evaluate the effects of IFN alpha and EV71 on miR-362-3p expression. Antiviral activity was assessed by modulating miR-362-3p levels. Compared to the NC mimic group, miR-362-3p in miR-362-3p mimic and IFN alpha treatment groups was increased;cell viability was enhanced;-lgTCID50 was reduced, and the replication of EV71 was inhibited. Further, VP1 mRNA and protein expression declined significantly in miR-362-3p mimic group vs NC mimic while they were notably elevated in the miR-362-3p inhibitor group vs NC inhibitor. IFN alpha treatment could induce miR-362-3p production and enhanced cell viability in HMC3 cells. Besides, when compared with the miR-362-3p mimic group, IFN alpha combined with miR-362-3p mimic group reduced -lgTCID50 and significantly decreased the expression of VP1 mRNA and protein. Conclusion: The inhibition of EV71-infected HMC3 cell replication was related to the upregulation of miR-362-3p induced by IFN alpha, which in turn enhanced antiviral viability.

关键词： Rotavirus   Lactose intolerance   PSTAT3   BALB/C suckling mice   IL-22  

摘要： Lactose intolerance (LI) is a significant contributor to diarrhea in infants and young children. Recent studies suggest a potential link between rotavirus (RV) infection and alterations in gut microbiota, which may play a role in the mechanism of LI. However, the precise underlying mechanism remains unclear and requires urgent clarification. In this study, we combined insights from immunology and gut microbiome research to propose that the activation of the IL-22/pSTAT3/RegIII gamma signaling pathway, triggered by gut microbiota, may be involved. To investigate this, we established a LI animal model using 7-day-old BALB/c mice, which were infected with RV via oral gavage. The syndromes of mice were carefully recorded and compared during the intervention experiment. Notably, we measured the the expression levels of immunocytokines and phosphorylated signaling proteins, including IL-22, phosphorylated STAT3, and RegIII gamma. Additionally, we assessed the relationships between gut microbiota and these key elements. Our results indicated that RV indeed causes LI, as severe diarrhea was observed in the mice during the first three days of RV infection, subsiding after seven days. ELISA results revealed an increase in IL-22 levels, phosphorylated STAT3, and RegIII gamma, suggesting that the mechanism of LI associated with RV is linked to the classical IL-22/pSTAT3/RegIII gamma signaling pathway. Furthermore, our analysis of the connections between gut microbes and this signaling pathway indicated that "Bacteroidetes" and Firmicutes were significantly positively correlated with IL-22 and lactase, respectively. This finding implies that alterations in gut microbiota may serve as a potential switch for the IL-22/pSTAT3/RegIII gamma signaling pathway, leading to increased lactase levels. In summary, our study demonstrated that the development of LI is associated with the activation of the IL-22/pSTAT3/RegIII gamma signaling pathway alongside changes in gut microbes during RV

关键词： Chronic bacterial prostatitis   Polygonum capitatum Buch.-ham. ex D. Don   Ciprofloxacin   Pik3cb   Gallic acid   Synergistic mechanism  

摘要： Ethnopharmacological relevance: Polygonum capitatum Buch.-Ham. ex D. Don (Pc, DB52/YC141-2003), a traditional herbal medicine utilized by the Miao nationality, is acknowledged for its therapeutic potential and efficacy in treating various urologic disorders, notably chronic bacterial prostatitis (CBP). Experimental and clinical studies have demonstrated that the combination of Pc with Ciprofloxacin (CIP) effectively alleviates the urinary symptoms in CBP patients, offering superior outcomes compared to monotherapy. However, the underlying mechanisms and specific constituents responsible for this synergistic effect remain poorly understood, which hinders its broader clinical application. Aim of the study: This study aims to elucidate the potential synergistic mechanism of Pc and CIP in ameliorating CBP and to identify the major active ingredient of Pc that contributed most significantly to the therapeutic efficacy when combined with CIP. Materials and methods: A rat model of CBP was induced in rats by prostate bilateral injections of Escherichia coli (E. coli.), followed by therapeutic intervention with a combination of Pc and CIP. The therapeutic effect of this combination was assessed by quantifying the Prostate viscera coefficient (PVC) and bacterial colonization. Histopathological changes in the prostate were observed using HE staining. The expressions of inflammatory mediators was quantified using Western blotting (WB), qRT-PCR, and immunohistochemical staining (IHC). To elucidate the molecular mechanisms underlying the synergism of the combination therapy, transcriptomic profiling was performed using RNA sequencing (RNA-seq). Differentially expressed genes were analyzed via Ingenuity Pathway Analysis (IPA) to identify regulated pathways, with critical targets further validated by qRTPCR and WB. For direct drug target identification, Tissue-thermal proteome profiling (Tissue-TPP) was implemented, incorporating differential temperature heat treatment, data-indepe

关键词： Triptolide   Salvianolic acid B   Immune regulation   Hepatic stellate cell   Th17/IL-17A axis  

摘要： Liver fibrosis represents a significant public health challenge, with immune dysregulation being a key pathological mechanism involved in its progression. The combined use of triptolide and salvianolic acid B has been demonstrated effective in ameliorating CCL4-induced liver fibrosis, but the underlying mechanism remains to be elucidated. In this study, we integrated network pharmacology and experimental validation to uncover the therapeutic mechanisms of the combined use of triptolide and salvianolic acid B. First, animal experiments demonstrated that the combination notably alleviated CCL4-induced liver injury and fibrosis. Second, network pharmacology was employed to predict the potential mechanisms, and the results highlighted Th17 cell differentiation and the IL-17 signaling pathway as the key pathways mediating the anti-fibrotic effects of the combination. Next, with the help of flow cytometry analysis, we confirmed that the combination effectively inhibited the differentiation of Th17 cells, both in vivo and in vitro, and decreased the expression of pro-inflammatory/ fibrotic cytokines. Finally, to preliminary ascertain the role of IL-17 signaling in HSC activation and if the combination could affect it, we used recombinant human IL-17A protein to trigger LX-2 cells, the results suggested that IL-17A signaling did engage in HSC activation, and the combination could significantly suppress IL17A-induced LX-2 activation partly by inhibiting the expression of IL-17RA. Taken together, those results implied that the combined use of triptolide and salvianolic acid B could attenuate CCL4-induce liver fibrosis, and this effect was related to the suppression of inflammation and hepatic stellate cell activation via Th17/IL-17A axis.