Immune Cells In Microenvironment Research Articles

A Dual-Function CD47-Targeting Nano-Drug Delivery System used to Regulate Immune and Anti-Inflammatory Activities in the Treatment of Atherosclerosis.

Atherosclerosis is a primary contributor to cardiovascular disease. Current studies have highlighted the association between the immune system, particularly immune cells, and atherosclerosis, although treatment options and clinical trials remain scarce. Immunotherapy for cardiovascular disease is still in its infancy. Bruton's tyrosine kinase (BTK), widely expressed in various immune cells, represents a promising therapeutic target for atherosclerosis by modulating the anti-inflammatory function of immune cells. In this study we introduced a polydopamine-based nanocarrier system to deliver the BTK inhibitor, ibrutinib, to atherosclerotic plaques with an active targeting property via an anti-CD47 antibody. Leveraging polydopamine's pH-sensitive reversible disassembly, the system offers responsive, controlled release within the pathologic microenvironment. This allows precise and efficient ibrutinib delivery, concurrently inhibiting the activation of the NF-κB pathway in B cells and the NLRP3 inflammasome in macrophages within the plaques. This treatment also modulates both the immune cell microenvironment and inflammatory conditions in atherosclerotic lesions, thereby conveying promising therapeutic effects for atherosclerosis in vivo. This strategy also provides a novel option for atherosclerosis treatment. This article is protected by copyright. All rights reserved.

Abstract NG07: Association between somatic microsatellite instability, hypermutation status, and specific T cell subsets in colorectal cancer tumors

Abstract NG07: Association between somatic microsatellite instability, hypermutation status, and specific T cell subsets in colorectal cancer tumors

Identification and validation of anoikis-related genes to clarify the prognosis and immune mechanisms of patients with low-grade glioma

BackgroundLow-grade glioma (LGG) has an extremely poor prognosis, and the mechanism leading to malignant development has not been determined. The aim of our study was to clarify the function and mechanism of anoikis and TIMP1 in the malignant progression of LGG. MethodsWe screened 7 anoikis-related genes from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases to construct a prognostic-predicting model. The study assessed the clinical prognosis, pathological characteristics, and immune cell infiltration in both high- and low-risk groups. Additionally, the potential modulatory effects of TIMP1 on proliferation, migration, and anoikis in LGG were investigated both in vivo and in vitro. ResultsIn this study, we identified seven critical genes, namely, PTGS2, CCND1, TIMP1, PDK4, LGALS3, CDKN1A, and CDKN2A. Kaplan‒Meier (K‒M) curves demonstrated a significant correlation between clinical features and overall survival (OS), and single-cell analysis and mutation examination emphasized the heterogeneity and pivotal role of hub gene expression imbalances in LGG development. Immune cell infiltration and microenvironment analysis further elucidated the relationships between key genes and immune cells. In addition, TIMP1 promoted the malignant progression of LGG in both in vitro and in vivo models. ConclusionsThis study confirmed that TIMP1 promoted the malignant progression of LGG by inhibiting anoikis, providing insights into LGG pathogenesis and potential therapeutic targets.

Tumor associated microglia/macrophages utilize GPNMB to promote tumor growth and alter immune cell infiltration in glioma

Tumor-associated microglia and blood-derived macrophages (TAMs) play a central role in modulating the immune suppressive microenvironment in glioma. Here, we show that GPNMB is predominantly expressed by TAMs in human glioblastoma multiforme and the murine RCAS-PDGFb high grade glioma model. Loss of GPNMB in the in vivo tumor microenvironment results in significantly smaller tumor volumes and generates a pro-inflammatory innate and adaptive immune cell microenvironment. The impact of host-derived GPNMB on tumor growth was confirmed in two distinct murine glioma cell lines in organotypic brain slices from GPNMB-KO and control mice. Using published data bases of human glioma, the elevated levels in TAMs could be confirmed and the GPNMB expression correlated with a poorer survival.

Anoikis-related gene signatures predict prognosis of lung adenocarcinoma patients and reveal immune infiltration.

Lung adenocarcinoma (LUAD), a type of lung cancer, is one of the most aggressive and deadly malignancies worldwide. Malignant tumor cells exhibit strong anti-anoikis properties to achieve distant metastasis through the circulatory system. However, more research is needed to understand how anoikis is involved in the progression, metastasis and especially the prognosis of LUAD. We obtained anoikis-related genes (ARGs) from two websites, Harmonizome and Genecards, and integrated them to select and model the genes associated with LUAD prognosis. In addition, we investigated differences in the immune cell microenvironment and pathways of enrichment analysis between subtypes. We finally constructed a nomogram based on ARGs and used decision curve analysis (DCA) to demonstrate that this model could help clinicians make clinical decisions. Sixty-four differentially expressed genes (DEGs) were found to be associated with survival, and of these, six were chosen to build a prognostic model. The time-dependent receiver operating characteristic (ROC) curves showed that the model had a satisfactory predictive ability. Enrichment analysis and immune microenvironment analysis revealed that the immune status and drug sensitivity of populations at high and low risk were different. We integrated the clinicopathological features of LUAD with the risk score to build the nomogram. The nomogram was shown to be a good predictor of short- and long-term survival in LUAD patients through DCA analysis. This new model based on six ARGs and nomograms in our study could help patients with LUAD develop personalized treatment plans.

Abstract 5331: Spatial resolution of immune cell lineages in the tumor microenvironment of plasma cell dyscrasias

Abstract Plasma cell dyscrasias (PCDs) are diseases of the hematologic system, with Multiple Myeloma as the most common disease. Less common PCDs include primary and secondary extramedullary plasmacytomas (EMP), which occur in soft tissues. EMPs are typically solitary and infrequent, often localized in the upper aerodigestive and gastrointestinal tracts, lung, and lymph nodes among other organs, and can convert to Multiple Myeloma. Secondary extramedullary plasmacytomas are often noted in advanced disease following multiple rounds of therapy and are commonly associated with poor prognosis. Studies aimed at gaining a better understanding of soft tissue EMPs, whether solitary plasmacytomas with undiagnosed multiple myeloma or in the context of recurrent or metastatic EMP disease, are limited. In this study, we employ a comprehensive approach using multiple modalities, including dissociated CITE-seq, spatial transcriptomics, and multiplexed spatial immuno-fluorescence imaging to interrogate the tumor and immune landscape in EMP disease. Importantly, the use of the same biomarker antibodies across these modalities provides a cross-data-framework for a deep contextual understanding of the immune and tumor cell organization and cell-to-cell signaling in EMP. Ultimately, this study provides new insights into patient-to-patient and tumor location variability, tumor and immune cell microenvironment heterogeneity, and possible future therapeutic strategies for EMP. Citation Format: Medbh A. Dillon, Lisa Arvidson, Cole G. Phalen, Julian R. Ishibashi, John B. Johanneson, Zachary J. Thomson, Peiyao A. Zhao, Jocelin Malone, Mackenzie S. Kopp, Matthew Norton, Gabriella Spang, Susan A. Ludmann, Adam K. Savage, Claire E. Gustafson, Marla Glass, Emma L. Kuan, Tao Peng, Lucas T. Graybuck, Xiao-jun Li, Troy R. Torgerson, Peter J. Skene, Ananda W. Goldrath, Samuel Jensen, Stephanie Anover-Sombke, Melinda L. Angus-Hill. Spatial resolution of immune cell lineages in the tumor microenvironment of plasma cell dyscrasias [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5331.

Revolutionising immune research with organoid-based co-culture and chip systems.

The intertwined interactions various immune cells have with epithelial cells in our body require sophisticated experimental approaches to be studied. Due to the limitations of immortalised cell lines and animal models, there is an increasing demand for human in vitro model systems to investigate the microenvironment of immune cells in normal and in pathological conditions. Organoids, which are self-renewing, 3D cellular structures that are derived from stem cells, have started to provide gap-filling tissue modelling solutions. In this review, we first demonstrate with some of the available examples how organoid-based immune cell co-culture experiments can advance disease modelling of cancer, inflammatory bowel disease and tissue regeneration. Then, we argue that to achieve both complexity and scale, organ-on-chip models combined with cutting-edge microfluidics-based technologies can provide more precise manipulation and readouts. Finally, we discuss how genome editing techniques and the use of patient-derived organoids and immune cells can improve disease modelling and facilitate precision medicine. To achieve maximum impact and efficiency, these efforts should be supported by novel infrastructures such as organoid biobanks, organoid facilities, as well as drug screening and host-microbe interaction testing platforms. All these together or in combination can allow researchers to shed more detailed, and often patient-specific, light on the crosstalk between immune cells and epithelial cells in health and disease.

SOX13 is a novel prognostic biomarker and associates with immune infiltration in breast cancer.

The transcription factor, SOX13 is part of the SOX family. SOX proteins are crucial in the progression of many cancers, and some correlate with carcinogenesis. Nonetheless, the biological and clinical implications of SOX13 in human breast cancer (BC) remain rarely known. We evaluated the survival and expression data of SOX13 in BC patients via the UNLCAL, GEPIA, TIMER, and Kaplan-Meier plotter databases. Immunohistochemistry (IHC) was used to verify clinical specimens. The gene alteration rates of SOX13 were acquired on the online web cBioportal. With the aid of the TCGA data, the association between SOX13 mRNA expression and copy number alterations (CNA) and methylation was determined. LinkedOmics was used to identify the genes that co-expressed with SOX13 and the regulators. Immune infiltration and tumor microenvironment evaluations were assessed by ImmuCellAI and TIMER2.0 databases. SOX13 correlated drug resistance analysis was performed using the GDSC2 database. Higher SOX13 expression was discovered in BC tissues in comparison to normal tissues. Moreover, increased gene mutation and amplification of SOX13 were found in BC. Patients with increased SOX13 expression levels showed worse overall survival (OS). Cox analysis showed that SOX13 independently served as a prognostic indicator for poor survival in BC. Further, the expression of SOX13 was also confirmed to be correlated with tumor microenvironment and diverse infiltration of immune cells. In terms of drug sensitivity analysis, we found higher expression level of SOX13 predicts a high IC50 value for most of 198 drugs which predicts drug resistance. The present findings demonstrated that high expression of SOX13 negatively relates to prognosis and SOX13 plays an important role in cancer immunity. Therefore, SOX13 may potentially be adopted as a biomarker for predicting BC prognosis and infiltration of immune cells.

Immune Cells in the Tumor Microenvironment of Soft Tissue Sarcomas.

Soft tissue sarcomas (STSs) are a rare heterogeneous group of malignant neoplasms characterized by their aggressive course and poor response to treatment. This determines the relevance of research aimed at studying the pathogenesis of STSs. By now, it is known that STSs is characterized by complex relationships between the tumor cells and immune cells of the microenvironment. Dynamic interactions between tumor cells and components of the microenvironment enhance adaptation to changing environmental conditions, which provides the high aggressive potential of STSs and resistance to antitumor therapy. Today, active research is being conducted to find effective antitumor drugs and to evaluate the possibility of using therapy with immune cells of STS. The difficulty in assessing the efficacy of new antitumor options is primarily due to the high heterogeneity of this group of malignant neoplasms. Studying the role of immune cells in the microenvironment in the progression STSs and resistance to antitumor therapies will provide the discovery of new biomarkers of the disease and the prediction of response to immunotherapy. In addition, it will help to initially divide patients into subgroups of good and poor response to immunotherapy, thus avoiding wasting precious time in selecting the appropriate antitumor agent.

Role of the immune system in liver transplantation and its implications for therapeutic interventions

AbstractLiver transplantation (LT) stands as the gold standard for treating end‐stage liver disease and hepatocellular carcinoma, yet postoperative complications continue to impact survival rates. The liver's unique immune system, governed by a microenvironment of diverse immune cells, is disrupted during processes like ischemia–reperfusion injury posttransplantation, leading to immune imbalance, inflammation, and subsequent complications. In the posttransplantation period, immune cells within the liver collaboratively foster a tolerant environment, crucial for immune tolerance and liver regeneration. While clinical trials exploring cell therapy for LT complications exist, a comprehensive summary is lacking. This review provides an insight into the intricacies of the liver's immune microenvironment, with a specific focus on macrophages and T cells as primary immune players. Delving into the immunological dynamics at different stages of LT, we explore the disruptions after LT and subsequent immune responses. Focusing on immune cell targeting for treating liver transplant complications, we provide a comprehensive summary of ongoing clinical trials in this domain, especially cell therapies. Furthermore, we offer innovative treatment strategies that leverage the opportunities and prospects identified in the therapeutic landscape. This review seeks to advance our understanding of LT immunology and steer the development of precise therapies for postoperative complications.

Cellular Deconvolution Reveals Unique Findings in Several Cell Type Fractions Within the Basal Cell Carcinoma Tumor Microenvironment

Introduction: Despite therapeutic advancements, locally advanced and metastatic basal cell carcinomas continue to carry poor prognoses and high recurrence rates. Current treatment options remain suboptimal due to limited efficacy and associated adverse events. The objectives of this study are to 1) characterize the basal cell carcinoma immune cell microenvironment and 2) identify novel therapeutic targets.
 Methods: Transcriptome data representing 25 basal cell carcinoma and 25 control tissue samples were obtained from the Gene Expression Omnibus. Cell type fraction estimates were derived by least-squares deconvolution. Population differences were determined by Mann-Whitney U test.
 Results: Most significantly, two deconvolution algorithms similarly observed greater B cell infiltration in tumor samples compared to normal tissue (P<0.0001).
 Conclusion: Importantly, the results of this study provide new insight into the basal cell carcinoma tumor microenvironment and nominate testable immune cell populations for future therapeutic discovery. Study limitations include sample size and applicable background prediction levels of bulk deconvolution tools.

Crosstalk between septic shock and venous thromboembolism: a bioinformatics and immunoassay analysis.

Herein, we applied bioinformatics methods to analyze the crosstalk between septic shock (SS) and venous thromboembolism (VTE), focusing on the correlation with immune infiltrating cells. Expression data were obtained from the Gene Expression Omnibus (GEO) database, including blood samples from SS patients (datasets GSE64457, GSE95233, and GSE57065) and VTE patients (GSE19151). We used the R package "limma" for differential expression analysis (p value<0.05,∣logFC∣≥1). Venn plots were generated to identify intersected differential genes between SS and VTE and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Enrichment analysis. The protein-protein interaction (PPI) network of intersected genes was constructed by Cytoscape software. The xCell analysis identified immune cells with significant changes in VTE and SS and correlated them with significant molecular pathways of crosstalk. Finally, we validated the mRNA expression of crosstalk genes by qPCR, while Matrix Metalloprotein-9 (MMP-9) protein levels were assessed through Western blotting (WB) and Immunohistochemistry (IHC) in human umbilical vein endothelial cells (HUVECs) and mice. In the present study, we conducted a comparison between 88 patients with septic shock and 55 control subjects. Additionally, we compared 70 patients with venous thromboembolism to 63 control subjects. Twelve intersected genes and their corresponding three important molecular pathways were obtained: Metabolic, Estrogen, and FOXO signaling pathways. The resulting PPI network has 194 nodes and 388 edges. The immune microenvironment analysis of the two diseases showed that the infiltration levels of M2 macrophages and Class-switched memory B cells were correlated with the enrichment scores of metabolic, estrogen, and FOXO signaling pathways. Finally, qPCR confirmed that the expression of MMP9, S100A12, ARG1, SLPI, and ANXA3 mRNA in the SS with VTE group was significantly elevated. WB and IHC experiments revealed that MMP9 protein was significantly elevated in the experimental group. Metabolic, estrogen, and FOXO pathways play important roles in both SS and VTE and are related to the immune cell microenvironment of M2 macrophages and Class-switched memory B cells. MMP9 shows promise as a biomarker for diagnosing sepsis with venous thrombosis and a potential molecular target for treating this patient population.

ScCURE identifies cell types responding to immunotherapy and enables outcome prediction

A deep understanding of immunotherapy response/resistance mechanisms and a highly reliable therapy response prediction are vital for cancer treatment. Here, we developed scCURE (single-cell RNA sequencing [scRNA-seq] data-based Changed and Unchanged cell Recognition during immunotherapy). Based on Gaussian mixture modeling, Kullback-Leibler (KL) divergence, and mutual nearest-neighbors criteria, scCURE can faithfully discriminate between cells affected or unaffected by immunotherapy intervention. By conducting scCURE analyses in melanoma and breast cancer immunotherapy scRNA-seq data, we found that the baseline profiles of specific CD8+ T and macrophage cells (identified by scCURE) can determine the way in which tumor microenvironment immune cells respond to immunotherapy, e.g., antitumor immunity activation or de-activation; therefore, these cells could be predictive factors for treatment response. In this work, we demonstrated that the immunotherapy-associated cell-cell heterogeneities revealed by scCURE can be utilized to integrate the therapy response mechanism study and prediction model construction.

Activation of immune evasion machinery is a part of the process of malignant transformation of human cells

Malignant transformation of human cells is associated with their re-programming which results in uncontrolled proliferation and in the same time biochemical activation of immunosuppressive pathways which form cancer immune evasion machinery. However, there is no conceptual understanding of whether immune evasion machinery pathways and expression of immune checkpoint proteins form a part of the process of malignant transformation or if they are triggered by T lymphocytes and natural killers (NK) attempting to attack cells which are undergoing or already underwent malignant transformation. To address this fundamental question, we performed experimental malignant transformation of BEAS-2B human bronchial epithelium cells and RC-124 non-malignant human kidney epithelial cells using bracken extracts containing carcinogenic alkaloid called ptaquiloside. This transformation led to a significant upregulation of cell proliferation velocity and in the same time led to a significant upregulation in expression of key immune checkpoint proteins – galectin-9, programmed death ligand 1 (PD-L1), indoleamine 2,3-dioxygenase (IDO1). Their increased expression levels were in line with upregulation of the levels and activities of HIF-1 transcription complex and transforming growth factor beta type 1 (TGF-β)-Smad3 signalling pathway. When co-cultured with T cells, transformed epithelial cells displayed much higher and more efficient immune evasion activity compared to original non-transformed cells. Therefore, this work resolved a very important scientific and clinical question and suggested that cancer immune evasion machinery is activated during malignant transformation of human cells regardless the presence of immune cells in microenvironment.

Abstract B006: Dissecting metabolic alterations of clear cell renal cell carcinomas one cell at a time

Abstract Pseudohypoxia-related metabolic alterations are known to exist in clear-cell renal cell carcinoma (ccRCC). Here we characterized single-cell metabolic pathways with the 10Xmultiome assay comparing metabolic and compositional shifts in tumor vs. normal-adjacent samples. We analyzed 57 tumor and 6 normal-adjacent samples from the Dartmouth Renal Tumors Biobank collected between 1994-2009. Samples were enzymatically disassociated and stored at -80 C until 10X multiome processing. RNA counts were extracted using Seurat, cell types were deconvolved using Azimuth, and 85 KEGG metabolic pathways were interrogated using scMetabolism scores. Linear mixed-effects models compared the metabolic scores in 10 cell lineages (juxtaglomerular apparatus, glomeruli, proximal tubule system, nephron loop, distal tubules, collecting ducts, lymphoid, myeloid, endothelia, and other stroma cells) between tumor and normal adjacent adjusting for sex, patient age, stage and grade as fixed-effects and donor, and year of collection, as random-effects. P-values were calculated using Satterthwaite's method. The mean age at diagnosis was 61.7 yrs (SD: 12.5), and 67% were stages I/II. 79,804 tumor and 5,967 normal-adjacent cells were analyzed. 34,318 cells were nephron epithelia (juxtaglomerular, glomeruli, renal tubules), 5,947 were collecting ducts, 18,959 were immune, 13,445 were endothelial, and 13,112 were other stromal cells. Out of 85 KEGG metabolic pathways, nine were prioritized based on the kidney cancer literature; glycolysis, oxidative phosphorylation, TCA cycle, fatty acid biosynthesis, glutamine/glutamate, arachidonic acids, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and glutathione. When comparing vs. the corresponding normal adjacent cell type, in tumor cells: 1) glutamine/glutamate expression increased in lymphoid, endothelial, and stromal cells; 2) fatty acid biosynthesis increased in myeloid and reduced in lymphoid, collecting ducts-like, and nephron loop-like cells, 3) glutathione and glycolysis increased in glomeruli-like and nephron loops-like, 4) glycolysis increased in collecting ducts-like and endothelial, 5) oxidative phosphorylation and TCA cycle genes decreased in distal tubules-like, collecting ducts-like, and endothelia, 6) arachidonic acid metabolism increased in nephron loops-like and glomeruli-like, and 7) phenylalanine, tyrosine, and tryptophan biosynthesis decreased in nephron loops-like, distal tubules-like, and collecting ducts-like cells. Cell-specific metabolic shifts were observed between tumor and normal adjacent samples that cannot be explained only by sample heterogeneity or cell lineages. Oxidative phosphorylation and TCA cycle modifications were observed in tumor endothelia and cells resembling distal tubules and collecting ducts. Similarly, glutamate and fatty acid alterations were focused on tumor microenvironment immune cells. Further dissecting these findings will provide additional therapeutic avenues for newer targets, such as glutaminase inhibitors and combinations with current therapies. Citation Format: Lucas A. Salas, Ze Zhang, Laurent Perreard, Fred W. Kolling, Jason R. Pettus, Brock C. Christensen. Dissecting metabolic alterations of clear cell renal cell carcinomas one cell at a time [abstract]. In: Proceedings of the AACR Special Conference: Advances in Kidney Cancer Research; 2023 Jun 24-27; Austin, Texas. Philadelphia (PA): AACR; Cancer Res 2023;83(16 Suppl):Abstract nr B006.

Integrated bioinformatics analysis of differentially expressed genes in the temporomandibular joint internal derangement.

This study aimed to identify significant mechanisms and potential treatments for temporomandibular joint internal derangement (TMJD) through integrated bioinformatics analysis. Gene expression data sets (GSE66864) from the Gene Expression Omnibus (GEO) database were downloaded. Differentially expressed genes (DEGs) were identified both in the treatment groups and in controls by R packages. Network analysis of protein-protein interaction (PPI) and Human Protein Atlas was used to explore DEGs' potential function. DGIdb database was utilized to gain potential drug targets. In conclusion, 126 DEGs were selected for TMJD through bioinformatics analysis. Both GO and Kyoto Encyclopedia of Genes and Genomes analyses combined showed the pathways involved in TMJD. A PPI network was constructed to select the top 10 hub genes, of which five hub genes were chosen for further investigation. Moreover, the microenvironment of immune cells related to hub genes was evaluated by R packages. Macrophages play an important role in inflammation and oral-related tumors. The Human Protein Atlas analysis indicated that the five hub genes are highly related to head and neck cancer. Finally, eight potential drugs were selected for two genes using the DGIdb database. Our integrated bioinformatics analysis identified DEGs in TMJD and provided potential ideas for further research and treatment approaches. However, experimental validation of the hub genes and potential drug targets is still needed. The key mechanisms of the identified genes and their potential roles as biomarkers or drug targets in TMJD require further investigation.

229-LB: Generation of Human CAR-Treg Cells for Inducing Immunological Tolerance against Diabetogenic T Cells in Human Pancreatic Islets

We have developed pancreatic beta-cell, antigen-specific, Chimeric Antigen Receptor (CAR) T regulatory cells (Tregs) and explored their therapeutic potential against type 1 diabetes (T1D)/Latent Autoimmune Diabetes of Adult (LADA) in human pancreatic tissues ex-vivo. Two human Glutamic Acid Decarboxylase 65 kD isoform (GAD65) B-cell paratopes known to bind to two immunodominant regions in the N-terminal (CAR-N) and Middle (CAR-M) regions of human GAD65 were selected for assembly onto T cell receptors of Tregs. We first show that GAD65-CAR Tregs had a superior proliferation index (3 times than naïve Tregs) and were able to suppress diabetogenic, T cells in-vitro and this suppression was significantly superior to naïve Tregs at 2:1 (Tresp vs Treg) ratio (p&amp;lt;0.004). Further, in ex vivo co-culture experimentation with pancreatic islets, confocal images demonstrated homing of GAD65-CAR Tregs to pancreatic islets as early as 24 hrs. With live immunofluorescence imaging and flow cytometry we demonstrated that GAD65-CAR Tregs from T1D/LADA patient were able to suppress diabetogenic cytotoxic T lymphocytes (CTLs) (p&amp;lt;0.04) in the immune cell microenvironment in presence of heterologous pancreatic islet ex vivo. These findings verify that GAD65-CAR Tregs recognize endogenous GAD65 on isolated islets in an MHC-independent manner, are proliferative robustly and suppress autologous CTLs. Thus, GAD65-CAR Tregs can induce tolerance against T1D diabetogenic T cells and the use of GAD65-CAR Tregs can prove a promising strategy for inducing tolerance in islet transplantation to replace the immunosuppressive drugs in a safe potent manner. Finally, homing of the GAD65-CAR Tregs to the islets, provided evidence for antigen-specific re-directionality. The robust proliferation and superior suppression of diabetogenic T cells by GAD65-CAR Tregs validate the functionality of GAD65-CAR Tregs in inducing tolerance in T1D patients and host vs graft islet rejections. Disclosure S. Imam: None. S. Rafiqi: None. J. C. Jaume: None. Funding University of Toledo

Systematic pan-cancer analysis identifies cuproptosis-related gene DLAT as an immunological and prognostic biomarker.

Lipoylated dihydrolipoamide S-acetyltransferase (DLAT), the component E2 of the multi-enzyme pyruvate dehydrogenase complex, is one of the key molecules of cuproptosis. However, the prognostic value and immunological role of DLAT in pan-cancer are still unclear. Using a series of bioinformatics approaches, we studied combined data from different databases, including the Cancer Genome Atlas, Genotype Tissue-Expression, the Cancer Cell Line Encyclopedia, Human Protein Atlas, and cBioPortal to investigate the role of DLAT expression in prognosis and tumor immunity response. We also reveal the potential correlations between DLAT expression and gene alterations, DNA methylation, copy number variation (CNV), tumor mutational burden (TMB), microsatellite instability (MSI), tumor microenvironment (TME), immune infiltration levels, and various immune-related genes across different cancers. The results show that DLAT displays abnormal expression within most malignant tumors. Through gene set enrichment analysis (GSEA), we found that DLAT was significantly associated with immune-related pathways. Further, the expression of DLAT was also confirmed to be correlated with the tumor microenvironment and diverse infiltration of immune cells, especially tumor-associated macrophages (TAMs). In addition, we found that DLAT is co-expressed with genes encoding major histocompatibility complex (MHC), immunostimulators, immune inhibitors, chemokines, and chemokine receptors. Meanwhile, we demonstrate that DLAT expression is correlated with TMB in 10 cancers and MSI in 11 cancers. Our study reveals that DLAT plays an essential role in tumorigenesis and cancer immunity, which may be used to function as a prognostic biomarker and potential target for cancer immunotherapy.

Neurotransmitters arrive to control systemic autoimmunity

Immune cell microenvironment plays a major role in the aberrant function of immune cells in systemic lupus erythematosus. Zeng and co-authors show that in human and murine lupus, splenic stromal cell-derived acetylcholine switches B cell metabolism to fatty acid oxidation and promotes B cell autoreactivity and disease development.

Hemagglutinin expressed by yeast reshapes immune microenvironment and gut microbiota to trigger diverse anti-infection response in infected birds.

The H5N8 influenza virus is a highly pathogenic pathogen for poultry and human. Vaccination is the most effective method to control the spread of the virus right now. The traditional inactivated vaccine, though well developed and used widely, is laborious during application and more interests are stimulated in developing alternative approaches. In this study, we developed three hemagglutinin (HA) gene-based yeast vaccine. In order to explore the protective efficacy of the vaccines, the gene expression level in the bursa of Fabricius and the structure of intestinal microflora in immunized animals were analyzed by RNA seq and 16SrRNA sequencing, and the regulatory mechanism of yeast vaccine was evaluated. All of these vaccines elicited the humoral immunity, inhibited viral load in the chicken tissues, and provided partial protective efficacy due to the high dose of the H5N8 virus. Molecular mechanism studies suggested that, compared to the traditional inactivated vaccine, our engineered yeast vaccine reshaped the immune cell microenvironment in bursa of Fabricius to promote the defense and immune responses. Analysis of gut microbiota further suggested that oral administration of engineered ST1814G/H5HA yeast vaccine increased the diversity of gut microbiota and the increasement of Reuteri and Muciniphila might benefit the recovery from influenza virus infection. These results provide strong evidence for further clinical use of these engineered yeast vaccine in poultry.