Level Of Immunity Research Articles

Exploring the dual roles of sec-dependent effectors from Candidatus Liberibacter asiaticus in immunity of citrus plants.

The three SDEs of CLas were expressed in citrus leaves by AuNPs-PEI mediated transient expression system, and promoted the proliferation of CLas and inhibited citrus immunity. Huanglongbing (HLB) is the most severe bacterial disease of citrus caused by Candidatus Liberibacter asiaticus (CLas). CLas suppress host immune responses and promote infection by sec-dependent effectors (SDEs), thus insight into HLB pathogenesis is urgently needed to develop effective management strategies. In this study, we focused on the roles of SDE4310, SDE4435 and SDE4955 in citrus. We found that the expression of SDE4310, SDE4435 and SDE4955 to increase with increasing citrus immune genes CsPR1, CsPR2, CsPR5, CsNPR1, CsRBOHD, CsMAP3K and CsBIK1, suggesting that the level of citrus immunity could be judged by the expression of SDE. To further explore the relationship between these three SDEs and citrus immunity, we established a transient expression system in citrus leaves, using gold nanoparticle-polyethyleneimine (AuNPs-PEI) to deliver recombinant plasmid containing SDE4310, SDE4435 or SDE4955 respectively into citrus leaves. Results showed that SDE4310, SDE4435 and SDE4955 were successfully expressed in citrus leaves using this transient expression system, and found that SDE4310, SDE4435 and SDE4955 could promote the CLas proliferation by decreasing the immune gene expression of the citrus. Additionally, we used AuNPs-PEI to deliver siRNA4310 to citrus cells, significantly reducing the expression of SDE4310 within 3days. Although the suppression of SDE4310 expression did not inhibit the CLas proliferation, it increased the expression level of CsPR1, CsNPR1 and CsBIK1. This is also the first time that AuNPs-PEI has been found to be able to deliver exogenous plasmids into citrus cells and express the target protein, providing a new method for future studies on citrus HLB.

Limited evidence of association between dysregulated immune marker levels and telomere length in severe mental disorders.

Accelerated ageing indexed by telomere attrition is suggested in schizophrenia spectrum- (SCZ) and bipolar disorders (BD). While inflammation may promote telomere shortening, few studies have investigated the association between telomere length (TL) and markers of immune activation and inflammation in severe mental disorders. Leucocyte TL defined as telomere template/amount of single-copy gene template (T/S ratio), was determined in participants with SCZ (N = 301) or BD (N = 211) and a healthy control group (HC, N = 378). TL was analysed with linear regressions for associations with levels of 12 immune markers linked to SCZ or BD. Adjustments were made for a broad range of potential confounding variables. TL was measured by quantitative polymerase chain reaction (qPCR) and the immune markers were measured by enzyme immunoassays. A positive association between levels of soluble tumour necrosis factor receptor 1A (sTNF-R1) and TL in SCZ (β = 0.191, p = 0.012) was observed. Plasma levels of the other immune markers were not significantly associated with TL in the BD, SCZ or HC groups. There was limited evidence of association between immune markers and TL in SCZ and BD. The results provide little support for involvement of immune dysregulation, as reflected by current systemic markers, in telomere attrition-related accelerated ageing in severe mental disorders.

Characterization of the colostrum proteome of primiparous Holstein cows and its association with colostrum immunoglobulin G concentrations

BackgroundThe objective was to characterize the colostrum proteome of primiparous Holstein cows in association with immunoglobulin G (IgG) content. Immediately after calving, colostrum samples were collected from 18 cows to measure IgG concentration. Based on colostrum IgG content, samples were classified through cluster analysis and were identified as poor, average, and excellent quality. The proteome was assessed with quantitative shotgun proteomics; abundance data were compared among the colostrum types; enrichment analysis of metabolic processes and proteins classes was performed as well. We also tested correlations between this proteome and blood globulin level of cows and passive immunity level of calves.ResultsOn average, 428 proteins were identified per sample, which belonged mainly to cellular process, biological regulation, response to stimulus, metabolic process, and immune system process. Most abundant proteins were complement C3 (Q2UVX4), alpha-S1-casein (P02662), Ig-like domain-containing protein (A0A3Q1M032), albumin (A0A140T897), polymeric immunoglobulin receptor (P81265), lactotransferrrin (P24627), and IGHG1*01 (X16701_4). Colostrum of excellent quality had greater (P < 0.05) abundance of serpin A3-7 (A2I7N3), complement factor I (A0A3Q1MIF4), lipocalin/cytosolic fatty-acid binding domain-containing protein (A0A3Q1MRQ2), complement C3 (E1B805), complement component 4 binding protein alpha (A0AAF6ZHP5), and complement component C6 (F1MM86). However, colostrum of excellent quality had lower (P < 0.05) abundance of HGF activator (E1BCW0), alpha-S1-casein (P02662), and xanthine dehydrogenase/oxidase (P80457). This resulted in enrichment of the biological processes predominantly for complement activation alternative pathway, complement activation, complement activation classical pathway, humoral immune response, leukocyte mediated immunity, and negative regulation of endopeptidase activity in excellent-quality colostrum. Additionally, some colostrum proteins were found to be correlated with the blood globulin level of cows and with the passive immunity level of calves (P < 0.05; r ≥ 0.57).ConclusionsThis study provides new insights into the bovine colostrum proteome, demonstrating associations between IgG levels and the abundance of other proteins, as well as the enrichment of metabolic processes related to innate immune response. Thus, results suggest that the colostrum proteomic profile is associated with the content of IgG. Future research should deeply explore the association of these findings with pre-calving nutrition status and blood composition of the cow, and with passive immunity transfer to the calf.

Integrative Single-Cell and Bulk RNA Sequencing Identifies a Macrophage-Related Prognostic Signature for Predicting Prognosis and Therapy Responses in Colorectal Cancer.

Colorectal cancer (CRC) is one of the most common malignant tumors, characterized by a high incidence and mortality rate. Macrophages, as a key immune cell type within the tumor microenvironment (TME), play a key role in tumor immune evasion and the progression of CRC. Therefore, identifying macrophage biomarkers is of great significance for predicting the prognosis of CRC patients. This study integrates scRNA-seq and bulk RNA-seq data to identify macrophage-related genes in CRC. By applying a comprehensive machine learning framework, the macrophage-related prognostic signature (MRPS) was constructed by 15 macrophage-related genes with prognostic values. The MRPS demonstrated strong predictive performance across multiple datasets, effectively stratifying high-risk and low-risk patients in terms of overall survival (OS) and disease-specific survival (DSS). Furthermore, immune analysis revealed significant differences between the high-risk and low-risk groups in immune cell infiltration levels and immune checkpoint gene expression patterns. Drug screening identified several small molecules, including Bortezomib and Mitoxantrone, as potential therapeutic options for high-risk patients. Pseudotime trajectory analysis further highlighted the potential role of genes comprising the MRPS in macrophage differentiation. This study provides a powerful tool for personalized prognosis prediction in CRC patients, offering new insights into macrophage-driven mechanisms in tumor progression and potential therapeutic strategies.

Role of immune cell homeostasis in research and treatment response in hepatocellular carcinoma

Introduction Recently, immune cells within the tumor microenvironment (TME) have become crucial in regulating cancer progression and treatment responses. The dynamic interactions between tumors and immune cells are emerging as a promising strategy to activate the host's immune system against various cancers. The development and progression of hepatocellular carcinoma (HCC) involve complex biological processes, with the role of the TME and tumor phenotypes still not fully understood. Therefore, it is essential to investigate the importance of immune cell homeostasis in HCC. Additionally, understanding the molecular mechanisms and biological functions underlying tumor–immune cell interactions is increasingly recognized as vital for improving therapeutic outcomes in clinical settings. Methods A total of 790 HCC samples were selected from public databases and real-world independent clinical cohorts. Machine learning methods, focusing on immune-related indicators, were applied to these samples. The Boruta algorithm was employed to develop an ICI score, which was used to assess patient prognosis and predict responses to immunotherapy. Additionally, a new immune subtype analysis of HCC was performed. Cellular-level experiments confirmed the interaction between TME-related factors and the tumor microenvironment in HCC. To further validate the predictive power of the ICI score, a clinical cohort study was conducted at an independent clinical center. Results By evaluating immune gene expression levels, immune cell abundance, Immunescore, and Stromalscore, we initially identified three distinct immune subtypes of HCC, each showing significant differences in survival rates and heterogeneity. Subsequently, DEGs from 1022 immune subtypes were used to classify HCC samples into three immune genotypes, each characterized by distinct prognosis and tumor immune microenvironment (TIME) profiles. Furthermore, we developed the ICI score, a novel immunophenotyping method for HCC, which revealed significant variations based on gender, stage, progression, and DNA mutation profiles (p < 0.05). The ICI score also effectively predicted responses to immunotherapies, particularly through the chemokine signaling, focal adhesion, and JAK/STAT signaling pathways. Conclusion This research demonstrated that TME and immunophenotyping clusters can enhance prognostic accuracy for HCC patients. The independent prognostic indicators identified underscore the connection between tumor phenotype and the immune environment in HCC.

GPX8 as a potential prognostic marker in gastric and colorectal cancer.

Gastric and colorectal cancers are common malignancies with high incidence and mortality worldwide. Early detection and individualized treatment are crucial to improving patient outcomes. Glutathione peroxidase-8 (GPX8), a member of the glutathione peroxidase family, emerges as a potential target for intervention in the treatment of various cancers. This study investigated the potential of GPX8 as a prognostic marker in patients with gastric and colorectal adenocarcinoma. The study employed a multi-omics approach to analyze GPX8 expression in both tumor and adjacent normal tissue of stomach adenocarcinoma (STAD), colon adenocarcinoma (COAD) and rectum adenocarcinoma patients. The Cancer Genome Atlas database was used to download the microarray data of GPX8 and clinical information for cancer patients. The TIMER database and TNMplot database were used to systematically evaluate the association of GPX8 with tumor-infiltrating lymphocytes in adenocarcinoma. Immunohistochemistry is used to detect GPX8 expression in clinical tumors and adjacent normal tissues. Univariate Cox analysis was performed to explore the relationship between GPX8 expression, immune cell levels, and the prognosis in cancer patients. GPX8 was significantly upregulated in tumor tissue and was associated with a poor prognosis in STAD and COAD patients. Furthermore, high GPX8 expression was found to be correlated with a higher degree of CD4+ T cell-infiltrating in COAD and neutrophil-infiltrating in STAD, indicating that GPX8 may play a role in immune evasion in cancer progression. This study highlights the potential of GPX8 as a prognostic marker in STAD and COAD, providing valuable insight into the development of personalized treatment strategies for cancer patients.

Barcoded SARS-CoV-2 viruses define the impact of duration and route of exposure on the transmission bottleneck in a hamster model.

The transmission bottleneck, defined as the number of viruses shed from one host to infect another, is an important determinant of the rate of virus evolution and the level of immunity required to protect against virus transmission. Despite its importance, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission bottleneck remains poorly characterized. We adapted a SARS-CoV-2 reverse genetics system to generate a pool of >200 isogenic SARS-CoV-2 viruses harboring specific 6-nucleotide barcodes, infected donor hamsters with this pool, and exposed contact hamsters to paired infected donors, varying the duration and route of exposure. Following exposure, the nasal turbinates, trachea, and lungs were collected and the number of barcodes in each tissue was enumerated. We found that longer and more direct exposures increased the transmission bottleneck and that the upper airway is the primary source of transmitted virus in this model. Together, these findings highlight the utility of barcoded viruses as tools to rigorously study virus transmission.

Ubiquitination-related Gene UBTD1 Mediates Poor Prognosis of Colorectal Cancer and Affects Colorectal Cancer Cell Proliferation and Ferroptosis.

Colorectal cancer (CRC) is the third most common cancer worldwide, and its occurrence and progression are often regulated by genetic and hereditary factors. Ubiquitination and the associated ubiquitin-binding enzymes and ligases regulate the tumor microenvironment and antitumor immunity to mediate tumor pathogenesis and progression. In this study, we examined the molecular characteristics and immunomodulatory effects of ubiquitination-associated genes that mediate CRC prognosis. The ubiquitination-related gene ubiquitin domain-containing protein 1 (UBTD1) was identified using bioinformatics and single-cell analyses. Subsequently, the ability of UBTD1 to predict CRC prognosis and immune checkpoint correlation was analyzed, the potential drug telatinib targeting UBTD1 was explored, and the correlation between UBTD1 and ferroptosis was analyzed. The role of UBTD1 in CRC and ferroptosis was verified using immunohistochemistry, gene knockout, western blot, cell cloning, and immunofluorescence. UBTD1 was identified as a significant prognostic and predictive gene for CRC and was involved in regulating immune checkpoint levels and immune cell function of CRC patients with CRC. High UBTD1 expression was found to enhance the presence of immune checkpoints that induce immune escape and inhibit ferroptosis onset. Telatinib may be a potential therapeutic drug targeting UBTD1. Our study demonstrated that UBTD1 is a prognostic marker for CRC in the regulation of ubiquitination and the tumor immune microenvironment and may serve as a modulator of ferroptosis.

Characterization of liver, adipose, and fecal microbiome in obese patients with MASLD: links with disease severity and metabolic dysfunction parameters

BackgroundMetabolic dysfunction-associated steatotic liver disease (MASLD) encompasses a range of histological findings from the generally benign simple steatosis to steatohepatitis (MASH) which can progress to fibrosis and cirrhosis. Several factors, including the microbiome, may contribute to disease progression.ResultsHere, we demonstrate links between the presence and abundance of specific bacteria in the adipose and liver tissues, inflammatory genes, immune cell responses, and disease severity. Overall, in MASLD patients, we observed a generalized obesity-induced translocation of gut bacteria to hepatic and adipose tissues. We identified microbial patterns unique to more severely diseased tissues. Specifically, Enterococcus, Granulicatella, and Morganellaceae abundance is positively correlated with immune cell counts and inflammatory gene expression levels, and both genera are significantly enriched in MASH patients. Brevibacterium is enriched in adipose tissues of patients with liver fibrosis.ConclusionTogether, these results provide further insight into the microbial factors that may be driving disease severity.5gz9zWLTQ4ZzRXZ3CTt_7qVideo

Vaccination status and hepatitis A immunity in children: insights from a large-scale study in Turkey

BackgroundHepatitis A remains a public health concern, particularly in areas with suboptimal sanitation. Introduced in Turkey's immunization schedule in 2011, the vaccine has improved immunity; however, gaps persist, especially in older, unvaccinated children. This study examines the seropositivity rates and antibody levels in children across different vaccination statuses and age groups, and to identify gaps in immunity, particularly among children those born before the introduction of the hepatitis A vaccine in Turkey.MethodsData from 9,858 patients, collected between August 2019 and March 2024, were analyzed to evaluate the immunological response to the vaccine. Patients were categorized into four groups based on vaccination status: under-vaccinated (children 6–18 months old), single-dose vaccinated (children 18–24 months old), fully vaccinated (children over 24 months old born after March 1, 2011), and unvaccinated (born before March 1, 2011, when routine hepatitis A vaccination began). Seropositivity rates and antibody levels were measured and statistically analyzed.ResultsIn this study, a total of 9,858 pediatric patients were assessed, with 1.1% under-vaccinated, 3.4% single-dose vaccinated, 60.5% fully vaccinated, and 35% unvaccinated. Seropositivity rates reached 93% in the fully vaccinated group, compared to 83% in the single-dose group and 63% in the unvaccinated group. Antibody titers were significantly higher in the fully vaccinated group, with statistically significant differences in seropositivity between this group and the unvaccinated group (p < 0.001). Additionally, a notable decrease in antibody levels was observed in the unvaccinated group as age increased (r = -0.365, p < 0.001).ConclusionThese findings underscore the critical need for targeted interventions to close vaccination gaps, especially among older, unvaccinated children who exhibit lower immunity levels. These insights are crucial for enhancing vaccination outreach and improving public health measures against hepatitis A.

The Impact of the Lactation Period Gut Microbiota of Two Different Beef Cattle Breeds on Spring-Born Calves.

To investigate the impact of maternal microbiota during lactation in different beef cattle breeds on their own immune levels, milk quality, and the growth and development of their offspring, this study measured the immune parameters, intestinal microbiota diversity, and milk quality of Pingliang red cattle and Simmental cattle, and performed a correlation analysis with the growth and development of their offspring. Our study showed that during lactation, Pingliang red cattle had significantly higher IL-6 levels than Simmental cattle, while the latter exhibited higher levels of immune factors such as IgG, IgA, IgM, IL-1β, and TNFα. The analysis of the intestinal microbiota of lactating cows found that Pingliang red cattle were rich in Bacteroidetes and Fibrobacteres, while Simmental cattle had a higher proportion of Actinobacteria. This difference may be related to the different adaptation strategies in energy metabolism and immune regulation between the two breeds. In addition, the analysis of milk quality between different beef cattle breeds revealed significant differences in protein, acidity, milk fat, and total solids. The correlation analysis found that Alistipes_communis, Firmicutes_bacterium_CAG_110, Alistipes_communis, Paludibacter_propionicigenes, Alistipes_sp._58_9_plus, and Bacteroidales_bacterium_55_9 were associated with both milk quality and the growth of offspring calves, including body weight, body length, chest girth, and cannon circumference. In conclusion, this study provides new insights into the intestinal microbiota, immune characteristics, and their effects on offspring health in different beef cattle breeds, and has important implications for breed selection and management strategies in the livestock industry.

Upregulated PCAT-1 predicts poor prognosis and reduced immune cell infiltration in head and neck squamous cell carcinoma through the miR-145-5p / FSCN-1 axis.

LncRNA PCAT-1 is known to promote cancer proliferation, invasion, and metastasis. However, its significance in HNSCC is not fully understood. This research investigates how the PCAT-1 / miR-145-5p / FSCN-1 axis promote HNSCC. We analyzed the gene expression patterns on 238 fresh-frozen samples, comparing tumors with their normal adjacent tissues (NATs). HNSCC samples showed higher PCAT-1 and FSCN-1 expression compared to NATs (p < 0.001 and p < 0.001, respectively). In contrast, miR-145-5p expression was markedly downregulated compared to NATs (p < 0.001). Notably, ROC curve analysis revealed exceptional diagnostic power, with an AUC of 0.83 for PCAT-1, 0.95 for miR-145-5p, and 0.91 for FSCN-1. Pearson correlation analysis unveiled a significant positive correlation between PCAT-1 and FSCN-1 expression levels (r = 0.084, p < 0.001) and negative correlations between FSCN-1 and miR-145-5p (r = -0.710, p < 0.001) as well as between PCAT-1 and miR-145-5p (r = -0.759, p < 0.001). Distinct molecular profiles were observed in the levels of PCAT-1, miR-145-5p, and FSCN-1 between HPV (-) and HPV ( +) 16 and 18 genotypes (p = 0.007, p = 0.027, and p = 0.002). MiR-145-5p expression showed significant differences between HPV (-) and HPV ( +) other genotypes (p = 0.035). FSCN-1 expression showed notable distinctions between HPV ( +) 18 & 16 and HPV ( +) other genotypes (p = 0.031). Elevated levels of lncRNA PCAT-1 promote HNSCC through the miR-145-5p/FSCN-1 axis and are associated with poor prognosis and reduced immune cell infiltration levels.

The Rational Regional Schedule of Preventive Vaccinations

Relevance. The National vaccination schedule (NVS) of the Russian Federation includes 12 infections against 17–18 indicated in the Vaccination Schedules of most EU countries, and requires improvement in terms of expanding the number of infections and contingents subject to immunization, which is reflected in the «Strategy for the development of vaccine prevention of infectious diseases for the period up to 2035» and the roadmap for its implementation. Since the process of entry of new infections and new vaccines into NVS is quite «difficult» and economically costly (development of domestic vaccines, modernization of domestic enterprises, localization of production of foreign vaccines at Russian enterprises, formation of commitment of medical workers and public confidence in new immunobiological medicines), part of the tasks is to ensure public accessibility to vaccine prevention of infections not indicated in the The NVS can be successfully solved within the framework of regional vaccination schedules (RVS).Aims. To propose and interpret the concept of rational regional vaccination schedules, and also to outline the principles of its formation and assessment of the immediate and long-term prospects for the development of regional foundations of vaccination prevention.Results and discussion. RVS is an expanded version of NVS by including additional vaccine-controlled infections, taking into account the epidemic situation and the presence of high-risk groups of infection in the region. In modern conditions, RVS have been developed and are effectively used in a number of subjects of the Russian Federation, such as Moscow, Perm Krai, Sverdlovsk, Chelyabinsk, Tyumen Regions, Yamalo-Nenets Autonomous District, etc. The implementation of vaccine prophylaxis in the regions using this tool provides conditions for equal access of children and adults to modern highly effective vaccines, increases public awareness of the benefits and risks of vaccination and the level of population immunity, contributes to improving mechanisms for providing citizens with immunobiological drugs and the formation of epidemic well-being in the region based on a risk-based approach.Conclusions. In accordance with this paradigm, the authors introduce and interpret the concept of an «Rational regional vaccination schedules», define the basic principles of its formation and assess the short- and long-term prospects for the development of the regional foundations of vaccination.

Evaluation of Tumor-Infiltrating Leukocytes in Endolymphatic Sac Tumor.

Endolymphatic sac tumors (ELSTs), as rare low-grade neoplasms, are primarily treated with surgery. This study analyzes the characteristics of tumor-infiltrating leukocytes (TILs) in ELSTs and their relationships with clinical features to explore the potential for immunotherapy in ELSTs. Clinical data and tumor specimens of 10 ELSTs patients who underwent surgery were retrieved. Immune expression levels of CD3, CD4, CD8, and CD66b were assessed by immunohistochemical staining and expressed as the integral optical density (IOD). Hematoxylin and eosin (HE) staining were performed to define the tumor nest and stroma of ELSTs. There were no significant differences in the IOD of CD3, CD4, CD8, and CD66b between the tumor nest and stroma or between von Hippel-Lindau (VHL) and non-VHL patients. The IOD of CD3, CD4, CD8, and CD66b appeared relatively higher in patients with endolymphatic hydrops (EH) than those without. Additionally, CD4 expression in the tumor stroma was significantly higher in patients with EH (p = 0.0381). TILs were present in both ELSTs nest and stroma with significant individual heterogeneity observed among patients. Patients with VHL disease showed a similar immune pattern to those with sporadic ELSTs. Notably, the ELST-related EH may be associated with lymphocytes infiltration. Combined with surgical intervention, immunotherapy may serve as an effective adjuvant therapeutic strategy. 4 Laryngoscope, 2025.

Euglena gracilis-derived β-glucan ameliorates particulate matter (PM2.5)-induced airway inflammation by modulating nuclear factor kappa B, mitogen-activated protein kinase, and nuclear factor erythroid 2-related factor 2 signaling pathways in A549 cells and BALB/c mice.

This study aimed to investigate the effects of β-glucan derived from Euglena gracilis (EGB), an edible microalga, on particulate matter (PM2.5)-induced airway inflammation in A549 cells and BALB/c mice. EGB effectively suppressed the mRNA and protein levels of inflammatory cytokines (IL-6, IL-1β, TNF-α, IL-8) and mediators (iNOS, COX-2), while inhibiting the NF-κB and MAPK signaling pathways triggered by PM2.5 exposure and reducing nuclear NF-κB levels. Additionally, EGB decreased PM2.5-induced ROS production and increased the protein levels of NRF2 and HO-1, along with genes encoding antioxidant enzymes (catalase, GPx, SOD1), associated with elevated nuclear NRF2 levels. EGB reduced immune cell infiltration and inflammatory cytokine levels in BALF and serum, both of which increased by PM2.5 exposure. EGB also significantly increased alveolar numbers while decreasing the gene expression of MMP1/9/13. Furthermore, EGB suppressed PM2.5-induced bronchial thickening and collagen-1 deposition by downregulating TGF-β1 expression, and alleviated goblet cell hyperplasia and mucin production in lung tissues. These results suggest that EGB effectively reduces PM2.5-induced airway inflammation by suppressing NF-κB and MAPK signaling pathways, lowering pro-inflammatory cytokines, and activating the NRF2-HO-1 signaling pathway to enhance antioxidant enzyme expression. This study highlights the potential of EGB as an edible functional agent for controlling PM-related airway inflammation.

Exploring the Effects of Gracilaria lemaneiformis Polysaccharides on the Fecal Microbiota and Fecal Metabolites of Fattening Pigs Based on 16S rDNA and Metabolome Sequencing.

Sixty Duroc × (Landrace × Yorkshire) castrated male finishing pigs were randomly divided into negative control (NC) and gracilaria lemaneiformis polysaccharides (GLP) groups to study the effects of GLP on the fecal microbiota and fecal metabolites of fattening pigs. The NC group was fed a basic diet, and the GLP group was fed a basic diet supplemented with 0.1% GLP. The diversity of the species and the fecal metabolites were analyzed using 16S rDNA sequencing and metabolome sequencing, respectively. The serum immune and antioxidant indices were analyzed using the ELISA assay. The abundances of hazardous bacteria such as Proteobacteria at the phylum level and Shigella at the genus level were extremely significantly decreased (p < 0.01) and those of beneficial bacteria such as Firmicutes at the phylum level (p < 0.01), Clostridium at the genus level (p < 0.01), and Lactobacillus at the genus level (p < 0.05) were significantly increased in the GLP group compared with the NC group. A total of 41 differentially expressed metabolites were identified. The expression of anti-inflammatory and antioxidant active substances, such as methyl cinnamate, protopanaxatriol, and isovanillic acid, was elevated in the GLP group. The ELISA assay showed increased GSH-Px activity (p < 0.01), T-AOC (p < 0.01), IgG (p < 0.01), IgA (p < 0.05), and IgM (p < 0.05) in the GLP group. These results indicate that dietary GLP supplementation can improve the antioxidant ability, anti-inflammatory ability, and immune level of fattening pigs by regulating fecal flora and metabolites and could be used as a functional feed additive.

Identification of a novel disulfidptosis-related gene signature in osteoarthritis using bioinformatics analysis and experimental validation

Osteoarthritis (OA) is a degenerative bone disease characterized by the destruction of joint cartilage and synovial inflammation, involving intricate immune regulation processes. Disulfidptosis, a novel form of programmed cell death, has recently been identified; however, the effects and roles of disulfidptosis-related genes (DR-DEGs) in OA remain unclear. We obtained six OA datasets from the GEO database, using four as training sets and two as validation sets. Differential expression analysis was employed to identify DR-DEGs, and unique molecular subtypes of OA were constructed based on these DR-DEGs. Subsequently, the immune microenvironment of OA patients was comprehensively analyzed using the “CIBERSORT” algorithm for immune infiltration. Various machine learning algorithms were utilized to screen characteristic DR-DEGs, and nomogram models and ROC curves were built based on these genes. The scRNA dataset (GSE169454) was used to classify chondrocytes in OA samples into distinct cell types, further exploring the gene distribution and correlation of characteristic DR-DEGs with specific cell subpopulations. Moreover, the expression levels of four characteristic DR-DEGs were validated through OA cell models and rat models. In our study, we identified 10 DR-DEGs with significant differences in expression within OA samples. Based on these DR-DEGs, two distinct molecular subtypes were recognized (cluster 1 and 2). ZNF484 and NDUFS1 were found to be significantly overexpressed in subtype 1, while the infiltration abundance of activated mast cells was markedly elevated in subtype 2. Moreover, significant differences were observed in the infiltration proportions of 11 immune cell types between OA and control samples, with 9 DR-DEGs demonstrating substantial correlations with immune cell infiltration levels. Further analysis of the scRNA dataset revealed that SLC3A2 and NDUFC1 were predominantly expressed in the preHTC subpopulation. All 10 DR-DEGs exhibited notably higher expression in the EC subpopulation across various cell types. The proportion of EC subgroups with high SLC3A2 expression increased, mainly enriching pathways related to inflammation, such as the IL-17 signaling pathway and TGF-beta signaling pathway. Using machine learning, we identified four characteristic DR-DEGs, which, in combination with the nomogram and ROC models, demonstrated promising performance in the diagnosis of OA. Additionally, in vivo validation confirmed a significant elevation of PPM1F expression in OA models. This study identified DR-DEGs as potential biomarkers for the diagnosis and classification of OA and provided a preliminary understanding of their role in the immune microenvironment. However, further experimental and clinical studies are required to validate their diagnostic value and therapeutic potential.

Circulating immune biomarkers correlating with response in patients with metastatic renal cell carcinoma on immunotherapy.

As multiple front-line immune checkpoint inhibitor (ICI)-based combinations are approved for metastatic renal cell carcinoma, biomarkers predicting for ICI responses are needed past clinical prognostication scores and transcriptome gene expression profiling. Circulating markers represent opportunities to assess baseline and dynamic changes in immune cell frequency and cytokine levels while on treatment. We conducted an exploratory prospective correlative study of 33 patients with metastatic clear cell renal cell carcinoma undergoing treatment with ICIs and correlated changes in circulating immune cell subsets and cytokines with clinical responses to treatment. Cell frequencies and cytokine levels were compared between responders and non-responders using unpaired parametric t tests, using a pre-specified level of significance of p<0.05. Classical monocyte subsets (CD14+ CD16-), as well as seven cytokines (IL-12/23 p40, macrophage inflammatory protein-1a, macrophage inflammatory protein-1b, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, IL-8, and TNF-alpha) were higher at baseline for responding versus non-responding patients. Dynamic changes in thymus- and activation-regulation chemokine (TARC), placental growth factor (PlGF), and vascular endothelial growth factor (VEGF) also correlated with patients with ICI response. In summary, macrophage activating agents were observed to be important in ICI response and may highlight the importance of the innate immune response in ICI responses.

Integration of machine learning and experimental validation to identify the prognostic signature related to diverse programmed cell deaths in breast cancer.

Programmed cell death (PCD) is closely related to the occurrence, development, and treatment of breast cancer. The aim of this study was to investigate the association between various programmed cell death patterns and the prognosis of breast cancer (BRCA) patients. The levels of 19 different programmed cell deaths in breast cancer were assessed by ssGSEA analysis, and these PCD scores were summed to obtain the PCDS for each sample. The relationship of PCDS with immune as well as metabolism-related pathways was explored. PCD-associated subtypes were obtained by unsupervised consensus clustering analysis, and differentially expressed genes between subtypes were analyzed. The prognostic signature (PCDRS) were constructed by the best combination of 101 machine learning algorithm combinations, and the C-index of PCDRS was compared with 30 published signatures. In addition, we analyzed PCDRS in relation to immune as well as therapeutic responses. The distribution of genes in different cells was explored by single-cell analysis and spatial transcriptome analysis. Potential drugs targeting key genes were analyzed by Cmap. Finally, the expression levels of key genes in clinical tissues were verified by RT-PCR. PCDS showed higher levels in cancer compared to normal. Different PCDS groups showed significant differences in immune and metabolism-related pathways. PCDRS, consisting of seven key genes, showed robust predictive ability over other signatures in different datasets. The high PCDRS group had a poorer prognosis and was strongly associated with a cancer-promoting tumor microenvironment. The low PCDRS group exhibited higher levels of anti-cancer immunity and responded better to immune checkpoint inhibitors as well as chemotherapy-related drugs. Clofibrate and imatinib could serve as potential small-molecule complexes targeting SLC7A5 and BCL2A1, respectively. The mRNA expression levels of seven genes were upregulated in clinical cancer tissues. PCDRS can be used as a biomarker to assess the prognosis and treatment response of BRCA patients, which offers novel insights for prognostic monitoring and treatment personalization of BRCA patients.

Immunomodulatory insights of monoterpene glycosides in endometriosis: immune infiltration and target pathways analysis

Endometriosis is a complex gynecological condition characterized by abnormal immune responses. This study aims to explore the immunomodulatory effects of monoterpene glycosides from Paeonia lactiflora on endometriosis. Using the ssGSEA algorithm, we assessed immune cell infiltration levels between normal and endometriosis groups. Key targets were identified through differential expression analysis of the GSE51981 dataset. Potential immunomodulatory targets of Paeonia lactiflora compounds were identified through Venn diagram analysis, followed by enrichment and machine learning analyses. A nomogram was developed for predicting endometriosis, while molecular docking explored compound-target interactions. Significant differences in immune cell infiltration were observed, with increased CD8 T cells, cytotoxic cells, and others in endometriosis. Differential expression analysis identified 43 potential targets. Enrichment analysis highlighted pathways involved in immune and inflammatory responses. Machine learning identified SSTR5, CASP3, FABP2, and SYK as critical targets, contributing to a nomogram that demonstrated good predictive performance for endometriosis risk. Molecular docking revealed strong interactions between Paeoniflorigenone and CASP3. Our findings suggest that monoterpene glycosides have therapeutic effects on endometriosis by modulating key immune-related targets and pathways, providing a basis for further investigation into Paeonia lactiflora’s potential as a treatment for this condition.