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关键词： 肾移植   排斥反应   可溶性白细胞抗原G   细胞因子   主要组织相容性复合体  

摘要： 目的　探讨外周血可溶性白细胞抗原G(sHLA-G)联合免疫细胞因子在肾移植排斥反应鉴别诊断中的应用价值。方法　本研究为病例对照研究。纳入2020年4月－2023年12月因血清肌酐升高在海军军医大学第二附属医院器官移植科住院的81例肾移植患者进行回顾性分析，其中诊断急性排斥反应32例(设为急性排斥组)，慢性排斥反应29例(设为慢性排斥组)，非排斥原因引起的肌酐升高20例(设为非排斥组)。选取同期住院和门诊随访血清肌酐正常且稳定的50例肾移植患者作为对照组。收集患者的临床资料，包括性别、年龄、血清肌酐、估算肾小球滤过率(eGFR)和尿蛋白阳性比例等，采集外周血并采用酶联免疫吸附法(ELISA)检测血浆sHLA-G及免疫细胞因子[γ干扰素(IFN-γ)、肿瘤坏死因子-β(TNF-β)、白细胞介素(IL)-2、IL-4、IL-10、IL-5、IL-6、IL-17]水平。将各组和全体肾移植患者按性别进行分层，比较sHLA-G水平的差异。结果　与对照组比较，急性排斥组、慢性排斥组、非排斥组血清肌酐水平和尿蛋白阳性比例明显升高，eGFR明显降低，且慢性排斥组和非排斥组血清肌酐水平高于急性排斥组，eGFR低于急性排斥组，差异均有统计学意义(P<0.05)。急性排斥组、慢性排斥组、非排斥组、对照组在性别、年龄、血型、体重指数、移植时间和免疫抑制剂使用情况等方面差异均无统计学意义(P>0.05)。急性排斥组、慢性排斥组患者的血浆sHLA-G水平均明显低于对照组[(19.665±11.233) U/mL vs. (24.785±21.668) U/mL vs. (44.918±39.898) U/mL，P<0.05]。慢性排斥组患者的sHLA-G/IL-2比值明显高于急性排斥组(5.844±6.248 vs. 1.825±1.574，P<0.05)，非排斥组患者的sHLA-G/IFN-γ比值明显高于慢性排斥组(3.452±3.283 vs. 1.543±2.03，P<0.05)。131例肾移植患者中，女性sHLA-G水平明显高于男性(P<0.05)。各组内比较，慢性排斥组女性sHLA-G水平明显高于男性(P<0.05)；急性排斥组、非排斥组和对照组女性sHLA-G水平虽高于男性，但差异无统计学意义(P>0.05)。结论　外周血sHLA-G水平与肾移植排斥反应存在相关性。sHLA-G/IL-2比值在肾移植急性与慢性排斥反应，sHLA-G/IFN-γ比值在肾移植慢性排斥反应与非排斥反应致肌酐升高的诊断和鉴别诊断中具有潜在的应用价值。

关键词： CD82蛋白   单克隆抗体   原核表达   流式细胞术  

摘要： 为研究CD82蛋白在猪病中的功能，提供一种针对猪CD82蛋白的单克隆流式抗体，本研究通过原核表达系统制备猪CD82蛋白大细胞外环结构域(EC2)重组抗原，经Ni-NTA亲和层析柱纯化后免疫BALB/c小鼠。通过4次免疫和ELISA筛选，获得抗体效价最高的小鼠，用其脾细胞进行细胞融合。对融合细胞上清用流式细胞术进行筛选，选取效果最好的1株杂交瘤细胞株11G3制备腹水并纯化，其抗体亚型为IgG2a型。通过间接ELISA和流式细胞术对纯化后的CD82 mAb进行验证，显示出高亲和力和特异性，能有效标记表达CD82蛋白的猪肺巨噬细胞（PAM）。本研究结果为CD82蛋白在猪病中的功能研究提供了重要的生物学工具，为进一步探索其在免疫调节和抗病毒反应中的作用奠定了基础。

关键词： Epigenomics   metabolic dysfunction-associated steatotic liver disease   lipid metabolism   inflammation  

摘要： ObjectiveEpigenetic alterations are critical regulators in the progression of metabolic dysfunction-associated steatotic liver disease (MASLD);however, the dynamic epigenomic landscapes are not well defined. Our previous study found that H3K27ac and H3K9me3 play important roles in regulating lipid metabolic pathways in the early stages of MASLD. However, the epigenomic status in the inflammation stages still needs to be ***57BL/6 male mice were fed with the methionine- and choline-deficient (MCD) or normal diet, and their serum and liver samples were collected after 6 weeks. Serum alanine aminotransferase (ALT), aspartate amino transferase (AST), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels were measured. Chromatin immunoprecipitation sequencing (ChIP-Seq) for H3K27ac and H3K9me3 was performed together with RNA sequencing (RNA-seq) and key regulators were *** target genes of enhancers with increased H3K27ac and decreased H3K9me3 signals are enriched in lipid metabolism and immuno-inflammatory pathways. Il-34 and Slc7al are identified as potential regulators in *** study reveals that active enhancers and heterochromatin associated with metabolic and inflammatory genes are extensively reprogrammed in MCD-diet mice, and Il-34 and Slc7al are potentially key genes regulating the progression of MASLD.

关键词： Macrophage   RNA-seq   direct current electric field (dcEF)   molecular dynamics (MD)  

摘要： Wearable devices have led to a growing interest in the effects of physical stimuli, such as electric fields (EF), on cells. However, the impact of EF on macrophages remains largely unexplored. To address this gap, we developed an electric-field chamber that applies direct current electric field (dcEF) to RAW 264.7 macrophages and analyzed the transcriptome changes and IL-1 beta protein conformation dynamics. Our results showed that exposure to dcEF did not affect cell morphology but significantly reduced cell viability. RNA-seq analysis revealed that dcEF altered the expression of 3518 upregulated and 2,531 downregulated genes. We also identified 6 hub genes using PPI network analysis and qPCR test. In addition, WB results suggested that dcEF increased the expression level of IL-1 beta. Next, we used molecular dynamics (MD) simulations to investigate the impact of dcEF on the conformation of the IL-1 beta protein. Our analysis results indicate that the dcEF's intensity significantly impacts various aspects of protein behavior, including its structure, motion, hydrogen bonding, and dipole moment. The alpha-helix structure exhibited the earliest degradation when exposed to an electric field, destroying other natural structures. In addition, the results of interleukin-1 (IL-1) gene family analysis suggest that this gene family is evolutionarily highly conserved. Overall, our findings provide insights into the influence of dcEF on macrophages and suggest potential applications of dcEF in cell biology and medicine.

关键词： reverse-bound peptides   HLA II   homology modeling   peptide-MHC   MHC II  

摘要： Recent discoveries have transformed our understanding of peptide binding in Major Histocompatibility Complex (MHC) molecules, showing that peptides, for some MHC class II alleles, can bind in a reverse orientation (C-terminus to N-terminus) and can still effectively activate CD4+ T cells. These finding challenges established concepts of immune recognition and suggests new pathways for therapeutic intervention, such as vaccine design. We present an updated version of PANDORA, which, to the best of our knowledge, is the first tool capable of modeling reversed-bound peptides. Modeling these peptides presents a unique challenge due to the limited structural data available for these orientations in existing databases. PANDORA has overcome this challenge through integrative modeling using algorithmically reversed peptides as templates. We have validated the new PANDORA feature through two targeted experiments, achieving an average backbone binding-core L-RMSD value of 0.63 & Aring;. Notably, it maintained low RMSD values even when using templates from different alleles and peptide sequences. Our results suggest that PANDORA will be an invaluable resource for the immunology community, aiding in the development of targeted immunotherapies and vaccine design.

关键词： HLA class I alleles   HLA class II alleles   chronic HBV infection   HBV clearance   HBV virus  

摘要： Chronic hepatitis B is linked with considerable liver-related morbidity and mortality globally. Human leukocyte antigen (HLA) polymorphisms affect the susceptibility and outcome of many immune-mediated diseases and infections. Our aim was to study the impact of HLA alleles on HVB-infected individuals in a Greek population. In total, 107 patients with chronic HBV infection (cHBV group) and 101 with spontaneous clearance (SC-group) of hepatitis B surface antigen (HBsAg) were genotyped for HLA-A, HLA-B, HLA-C, HLA-DRB1, and HLA-DQB1 loci by single-specific primer polymerase chain reaction (PCR-SSP). The HLA alleles' frequencies were compared between the two patient groups and healthy individuals from the North Greece Bone Marrow Donor Registry (14506 samples). We found a significantly increased frequency of HLA-C*01 and HLA-DRB1*16 alleles in the cHBV group versus the SC-group. The frequency of HLA-A*01, HLA-B*08, HLA-C*01, HLA-C*08, HLA-DRB1*03, and HLA-DQB1*05 alleles was significantly higher in cHBV patients versus healthy individuals, while the frequency of the HLA-B*38 allele was significantly lower. Our study showed an association of specific HLA alleles with either susceptibility or protection against chronic HBV infection.

关键词： oleocanthal   PAR-2   colorectal cancer   inflammation   TNF-alpha   calcium signaling  

摘要： Colorectal cancer (CRC) remains a principal contributor to oncological mortality worldwide, with chronic inflammation serving as a fundamental driver of its pathogenesis. Protease-activated receptor-2 (PAR-2), a G-protein-coupled receptor, orchestrates inflammation-driven tumorigenesis by potentiating NF-kappa B and Wnt/beta-catenin signaling, thereby fostering epithelial-mesenchymal transition (EMT), immune evasion, and therapeutic resistance. Despite its pathological significance, targeted modulation of PAR-2 remains an underexplored avenue in CRC therapeutics. Oleocanthal (OC), a phenolic constituent of extra virgin olive oil, is recognized for its potent anti-inflammatory and anti-cancer properties;however, its regulatory influence on PAR-2 signaling in CRC is yet to be elucidated. This study interrogates the impact of OC on PAR-2-mediated inflammatory cascades using HT-29 and Caco-2 CRC cell lines subjected to lipopolysaccharide (LPS)-induced activation of PAR-2. Expression levels of PAR-2 and TNF-alpha were quantified through Western blotting and RT-PCR, while ELISA assessed TNF-alpha secretion. Intracellular calcium flux, a pivotal modulator of PAR-2-driven oncogenic inflammation, was evaluated via Fluo-4 calcium assays. LPS markedly elevated PAR-2 expression at both mRNA and protein levels in CRC cells (p < 0.01, one-way ANOVA). OC administration (20-150 mu g/mL) elicited a dose-dependent suppression of PAR-2, with maximal inhibition at 100-150 mu g/mL (p < 0.001, Tukey's post hoc test). Concomitant reductions in TNF-alpha transcription (p < 0.01) and secretion (p < 0.001) were observed, corroborating the anti-inflammatory efficacy of OC. Additionally, OC ameliorated LPS-induced calcium dysregulation, restoring intracellular calcium homeostasis in a concentration-dependent manner (p < 0.01). Crucially, OC exhibited selectivity for PAR-2, leaving PAR-1 expression unaltered (p > 0.05), underscoring its precision as a therapeutic agent. These findings position

关键词： IL-33   ST2   carcinoma associated fibroblasts   regulatory T cells   head and neck squamous cell carcinoma   prognosis  

摘要： Background Interleukin-33 (IL-33) and Suppression of tumorigenicity 2 (ST2) expression are strongly associated with tumor growth and progression in diverse cancers, indicating the possibility of targeting the IL-33/ST2 axis pathway as a favorable therapeutic approach. However, the specific implications of IL-33/ST2 expression in Head and Neck Squamous Cell Carcinoma (HNSCC) prognosis are not fully understood. Thus, there is a need for more comprehensive research to verify the tasks and clinical significance of IL-33 and ST2 in HNSCC. Objectives The objective of this study was to evaluate the potential of differentially expressed IL-33 and ST2 in tumor tissues that could serve as novel biomarkers in HNSCC. Material & methods The Web of Science, Scopus, and PubMed electronic databases were searched and analyzed from January 2013 to July 2023. Results Nine studies fulfilling the inclusion criteria were analyzed. These selected studies were mainly having observational analytical study design, predominantly conducted within the Southeast Asian population. IL-33, primarily located in the stroma, demonstrates enhanced expression within carcinoma-associated fibroblasts (CAFs). Overexpression of IL-33 in CAFs correlates with its expression in tumor cells, as per some of these reports. Elevated IL-33 levels in CAFs are associated with unfavorable clinical outcomes. Increased IL-33 expression is related to poor nodal metastasis-free survival, indicating an adverse prognosis in HNSCC. In HNSCC, tumor cells and regulatory T cells (Tregs) expressed ST2. The degree of ST2 expression on Tregs corresponds to the abundance of IL-33 expressing CAFs. IL-33 increases the Tregs density and amplifies their suppressive capability. Poorer survival outcomes in HNSCC are linked to elevated ST2 expression in Tregs combined with the existence of IL-33-expressing CAFs. Conclusion CAF-driven cancer invasiveness relies on IL-33 signaling via paracrine and autocrine pathways. IL-33 may be a prognos

摘要： The catecholamine neurotransmitter dopamine is classically known for regulation of central nervous system (CNS) functions such as reward, movement, and cognition. Increasing evidence also indicates that dopamine regulates critical functions in peripheral organs and is an important immunoregulatory factor. We have previously shown that dopamine increases NF-kappa B activity, inflammasome activation, and the production of inflammatory cytokines such as IL-1 beta in human macrophages. As myeloid lineage cells are central to the initiation and resolution of acute inflammatory responses, dopamine-mediated dysregulation of these functions could both impair the innate immune response and exacerbate chronic inflammation. However, the exact pathways by which dopamine drives myeloid inflammation are not well defined, and studies in both rodent and human systems indicate that dopamine can impact the production of inflammatory mediators through both D1-like dopamine receptors (DRD1, DRD5) and D2-like dopamine receptors (DRD2, DRD3, and DRD4). Therefore, we hypothesized that dopamine-mediated production of IL-1 beta in myeloid cells is regulated by the ratio of different dopamine receptors that are activated. Our data in primary human monocyte-derived macrophages (hMDM) indicate that DRD1 expression is necessary for dopamine-mediated increases in IL-1 beta, and that changes in the expression of DRD2 and other dopamine receptors can alter the magnitude of the dopamine-mediated increase in IL-1 beta. Mature hMDM have a high D1-like to D2-like receptor ratio, which is different relative to monocytes and peripheral blood mononuclear cells (PBMCs). We further confirm in human microglia cell lines that a high ratio of D1-like to D2-like receptors promotes dopamine-induced increases in IL-1 beta gene and protein expression using pharmacological inhibition or overexpression of dopamine receptors. RNA-sequencing of dopamine-treated microglia shows that genes encoding functions in IL-1 be