Plant-Derived Extracellular-Vesicle-Like Particles: Immune Regulation and Disease Therapy

Flow cytometry is a widely used method in immunological research that allows for rapid and accurate analysis of individual cells to detect their immunophenotypic characteristics and functional status. This technique is particularly useful for studying immune cells. Flow cytometry can be used to detect and analyze various types of immune cells, such as T cells, B cells, dendritic cells, and macrophages. By analyzing the characteristic parameters of immune cells, such as killing activity, proliferative activity, and secretion factors, immune-related diseases can be diagnosed and treated. Flow cytometry can also be utilized to investigate various cell types within the immune system. For instance, it can be employed to study T lymphocytes in lymphoid tissue and B lymphocytes in bone marrow. This enables researchers to gain insights into the functions of these cells in immune regulation and the maintenance of internal environmental stability. Flow cytometry is of great significance in immunological research as it can be used not only to detect various types of immune cells but also to study different immune systems. In terms of immune regulation, flow cytometry can be used to detect and analyze various types of immune cells, such as B lymphocytes and macrophages, as well as their distribution and functional status in vivo. Flow cytometry can also be used to study subpopulations of T lymphocytes and B lymphocytes in bone marrow.

Key genes linking gut microbiota, immune cells, and osteoporosis: A multi-omics approach.

Emerging in vitro evidence suggests that histone modification plays a role in vitamin D‐mediated immune‐regulatory and anti‐inflammation effects. We hypothesized that DNA methylation is also involved in vitamin D deficiency mediated immune cell regulation. A genome‐wide methylation scan was performed using the Illumina HumanMethylation 27 BeadChip on leukocytes DNAs of 11 vitamin D deficient cases (serum 25(OH)D ≤25 nmol/L) and 11 age‐matched controls (serum 25(OH)D>75 nmol/L); the subjects were African American normal‐weight males aged 14–19 years. The Limma package was used to analyze each CpG site for differential methylation between cases and controls. To correct for multiple testing, the set of raw p values were converted to false discovery rates (FDR). Seventy nine CpG sites achieved statistical significance (raw p<0.001). The top two CpG sites (cg16317961 and cg04623955) survived multiple testing using a FDR of 10% (FDR=0.078). Moreover, half of the top ten genes have been implicated in immune cell activation, regulation and inflammatory response. In summary, our pilot data suggest that vitamin D deficiency is associated with methylation differences in blood leukocyte DNA in African American adolescents, suggesting that DNA methylation may be involved in vitamin D‐mediated immune regulation. Larger genome‐wide studies are warranted to confirm these findings.

Extracellular vesicles (EVs), secreted by all cellular organisms, are pivotal mediators of intercellular communication. By transporting biologically active cargos such as proteins, lipids, and nucleic acids, EVs facilitate transfer of molecular signals, effectively reflecting the characteristics of their parent cells. Immune cellderived EVs (iEVs) play a crucial role in the activation and regulation of both adaptive and innate immune responses. In the context of immune activation, iEVs drive immune cell development and activation, as well as enhance antigen presentation through both direct and cross-dressing mechanisms. Furthermore, iEVs act as signaling entities within immunological synapses, significantly amplifying immune response efficiency. In immune regulation, iEVs modulate the expression of immune checkpoint (IC) molecules and sustain immune homeostasis by transporting immunosuppressive cytokines and microRNAs, thereby mitigating excessive immune reactions. Nevertheless, the mechanistic underpinnings of iEV-mediated immune cell activation, antigen presentation, and immunoregulation remain inadequately explored. This review provides a comprehensive overview of the functions of iEVs from diverse immune cell origins and underlying mechanisms. It also examines cutting-edge engineering strategies targeting iEVs and their parent cells, while discussing their promising applications in oncology and immune-related diseases. These insights lay the foundation for the rational development of next-generation immunotherapies. While promising, the clinical translation of iEVs is hindered by low yield, high batch-to-batch variability, and insufficient targeting efficiency. The final section discusses key challenges and potential solutions.

Disabled-2 (DAB2) is a clathrin and cargo binding endocytic adaptor protein recognized for its multifaceted roles in signaling pathways involved in cellular differentiation, proliferation, migration, tumor suppression, and other fundamental homeostatic cellular mechanisms. The requirement for DAB2 in the canonical TGFβ signaling in fibroblasts suggested that a similar mechanism may exist in immune cells and that DAB2 may contribute to immunological tolerance and suppression of inflammatory responses. In this review, we synthesize the current state of knowledge on the roles of DAB2 in the cells of the innate and adaptive immune system, with particular focus on antigen presenting cells (APCs; macrophages and dendritic cells) and regulatory T cells (Tregs). The emerging role of DAB2 in the immune system is that of an immunoregulatory molecule with significant roles in Treg-mediated immunosuppression, and suppression of TLR signaling in APC. DAB2 itself is downregulated by inflammatory stimuli, an event that likely contributes to the immunogenic function of APC. However, contrary findings have been described in neuroinflammatory disorders, thus suggesting a highly context-specific roles for DAB2 in immune cell regulation. There is need for better understanding of DAB2 regulation and its roles in different immune cells, their specialized sub-populations, and their responses under specific inflammatory conditions.

Sepsis remains the leading cause of death of patients in pediatric intensive care unit.The adaptive immune dysfunction affects the occurrence and development of sepsis.Recent studies have found that interleukin-7 plays a critical part in the immune cell activation, proliferation and maturation, and it may have important means in the immune reconstitution in sepsis.Immunosuppression, the biological function of interleukin-7, the immune cell regulation and its application progress in sepsis were reviewed in this article. Key words: Sepsis; Interleukin-7; Immune regulation; Children

PurposeRheumatoid arthritis (RA) affects millions of people worldwide and causes chronic joint pain with incompletely understood pathogenesis and challenging management. Myosin light chain 1 (MYL1) a muscle regulatory protein, has an unknown role in RA pathogenesis.MethodsWe analyzed transcriptomic data from RA patients and healthy controls in the Gene Expression Omnibus (GEO). Using least absolute shrinkage and selection operator (LASSO) regression and random forest, we identified differentially expressed genes (DEGs). Diagnostic performance was assessed using receiver operating characteristic (ROC) curves and area under the curve (AUC). To explore potential mechanisms, we conducted functional enrichment and immune-infiltration analyses. We further validated MYL1 expression in a rat RA model using qRT-PCR and Western blotting.ResultsWe found MYL1 significantly downregulated in RA with high diagnostic value (AUC > 0.8). Enrichment analyses revealed its involvement in muscle structure development, immune regulation, and calcium signaling pathways. CIBERSORT analysis indicated associations with immune cell infiltration, particularly regulatory T cells, activated natural killer (NK) cells, and M1 macrophages. The rat model confirmed reduced MYL1 expression at both mRNA (p < 0.001) and protein (p = 0.009) levels, consistent with human data.ConclusionMYL1 is consistently downregulated in RA and may serve as a potential diagnostic biomarker. The results indicate that MYL1 may be involved in the pathological process of RA through calcium signaling, muscle function, and immune cell regulation. However, further clinical and mechanistic studies are warranted.

Immune metabolism is an emerging highlight in recent years. Revealing the metabolic characteristics of different immune cells in different responses may provide new perspective and direction for the pathogenesis and therapy of many immune-related diseases. Sepsis is a complex systemic inflammation caused by trauma, infection and other pathogenic factors. The immune cells have different metabolic features at different stages of the disease. These metabolic features are also involved in the regulation of immune cell proliferation, differentiation and function. By summarizing and analyzing the relevant literatures of immune cell metabolism and inflammation regulation in recent years, the metabolic regulatory factors of different immune cell subgroups and the related characteristics of immune cell metabolism in patients with sepsis were summarized. The in-depth understanding of the metabolic state in different immune cells, and the pathophysiological mechanism of septic immune disorders, especially the immune paralysis stage, would provide a theoretical basis for the clinical application of immune metabolic therapy.

Although mast cells play a critical role in allergic reactions, the cells are also involved in the protective immunity in the body. This study aims to investigate the role of mast cells in immune regulation during aberrant T helper (Th)2 responses. In this study, an adoptive antigen-specific Th2 response model was established with mast cell-deficient mice to test the role of mast cell in the immune regulation. Cell culture was employed to test the role of mast cells in the modulation of the expression of B cell lymphoma 6 protein (Bcl-6) in Th2 cells. The results showed that after adoptive transfer with immune cells, the mast cell-deficient mice showed stronger Th2 pattern responses in the intestine than that in the mast cell-sufficient mice. Mast cell-derived mouse mast cell protease-6 increased the expression of Bcl-6 in Th2 cells. Bcl-6 inhibited the expression of GATA-3 in Th2 cells, subsequently, forkhead box P3 was increased and the Th2 cytokines were reduced in the cells; the cells thus showed the immune regulatory properties similar to regulatory T cells. We conclude that bedsides initiating immune inflammation, mast cells also contribute to the immune regulation on Th2 polarization.

Wnt signaling is one of the important pathways to play a major role in various biological processes, such as embryonic stem-cell development, tissue regeneration, cell differentiation, and immune cell regulation. Recent studies suggest that Wnt signaling performs an essential function in immune cell modulation and counteracts various disorders. Nonetheless, the emerging role and mechanism of action of this signaling cascade in immune cell regulation, as well as its involvement in various cancers, remain debatable. The Wnt signaling in immune cells is very diverse, e.g., the tolerogenic role of dendritic cells, the development of natural killer cells, thymopoiesis of T cells, B-cell-driven initiation of T-cells, and macrophage actions in tissue repair, regeneration, and fibrosis. The purpose of this review is to highlight the current therapeutic targets in (and the prospects of) Wnt signaling, as well as the potential suitability of available modulators for the development of cancer immunotherapies. Although there are several Wnt inhibitors relevant to cancer, it would be worthwhile to extend this approach to immune cells.

LncRNAs exhibit subtype-specific expression, survival associations, and cancer-promoting effects in breast cancer

In recent years, B cells have emerged as an important player in the immune regulatory network. Using the animal model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE), we observed a lack of resolution of disease in B cell-deficient (μMT) mice. Whereas, their WT counterparts recovered, indicating a regulatory role for B cells in central nervous system (CNS) autoimmunity. CD4+Foxp3+ T regulatory cells (Tregs) also mediate resolution of EAE. Absence of CD80/CD86 (B7) on B cells correlated with late emergence of Tregs in the CNS during EAE, suggesting an interaction between B cells and Tregs through B7 molecules. Anti-CD20 mediated B cell depletion disrupted this interaction resulting in significant reduction in the number of Tregs and like μMT, manifested in unresolved EAE. Interestingly, in vitro studies identified a unique role for B cells in maintaining Treg Foxp3 expression, which was enhanced upon CD86 upregulation following anti-IgM activation. Adoptive transfer of B cells into μMT mice resulted in partial restoration of Treg numbers leading to resolution of EAE. When deficient in CD86, B cells had a significantly reduced ability to restore Treg numbers in μMT. These data identify a distinct role for B cells in immune regulation through CD86-mediated interaction with Tregs and maintenance of Foxp3. This novel immune regulatory property of B cells may be harnessed to develop therapies for autoimmune diseases.

Infectious bursal disease virus (IBDV) infection destroys the bursa of Fabricius, causing immunosuppression and rendering chickens susceptible to secondary bacterial or viral infections. IBDV large-segment-protein-expressing DNA has been shown to confer complete protection of chickens from infectious bursal disease (IBD). The purpose of the present study was to compare DNA-vaccinated chickens and unvaccinated chickens upon IBDV challenge by transcriptomic analysis of bursa regarding innate immunity, inflammation, immune cell regulation, apoptosis and glucose transport. One-day-old specific-pathogen-free chickens were vaccinated intramuscularly three times at weekly intervals with IBDV large-segment-protein-expressing DNA. Chickens were challenged orally with 8.2 × 10(2) times the egg infective dose (EID)50 of IBDV strain variant E (VE) one week after the last vaccination. Bursae collected at 0.5, 1, 3, 5, 7, and 10 days post-challenge (dpc) were subjected to real-time RT-PCR quantification of bursal transcripts related to innate immunity, inflammation, immune cell regulation, apoptosis and glucose transport. The expression levels of granzyme K and CD8 in DNA-vaccinated chickens were significantly (p < 0.05) higher than those in unvaccinated chickens upon IBDV challenge at 0.5 or 1 dpc. The expression levels of other genes involved in innate immunity, inflammation, immune cell regulation, apoptosis and glucose transport were not upregulated or downregulated in DNA-vaccinated chickens during IBDV challenge. Bursal transcripts related to innate immunity and inflammation, including TLR3, MDA5, IFN-α, IFN-β, IRF-1, IRF-10, IL-1β, IL-6, IL-8, iNOS, granzyme A, granzyme K and IL-10, were upregulated or significantly (p < 0.05) upregulated at 3 dpc and later in unvaccinated chickens challenged with IBDV. The expression levels of genes related to immune cell regulation, apoptosis and glucose transport, including CD4, CD8, IL-2, IFN-γ, IL-12(p40), IL-18, GM-CSF, GATA-3, p53, glucose transporter-2 and glucose transporter-3, were upregulated or significantly (p < 0.05) upregulated at 3 dpc and later in unvaccinated chickens challenged with IBDV. Taken together, the results indicate that the bursal transcriptome involved in innate immunity, inflammation, immune cell regulation, apoptosis and glucose transport, except for granzyme K and CD8, was not differentially expressed in DNA-vaccinated chickens protected from IBDV challenge.

In 2005, Altuvia and colleagues were the first to identify the gene that encodes miR-451 in the human pituitary gland, located in chromosome region 17q11.2. Subsequent studies have confirmed that miR-451 regulates various immune cells, including T cells, B cells, microglia, macrophages, and neutrophils, thereby influencing disease progression. The range of immune-related diseases affected encompasses various cancers, lymphoblastic leukemia, and injuries to the lungs and spinal cord, among others. Moreover, miR-451 is produced by immune cells and can regulate both their own functions and those of other immune cells, thus creating a regulatory feedback loop. This article aims to comprehensively review the interactions between miR-451 and immune cells, clarify the regulatory roles of miR-451 within the immune system, and assess its potential as both a therapeutic target and a biomarker for immune-related diseases.

BackgroundA systemic evaluation of immune cell infiltration patterns in experimental acute pancreatitis (AP) is lacking. Using multi-dimensional flow cytometry, this study profiled infiltrating immune cell types in multiple AP mouse models.MethodsThree AP models were generated in C57BL/6 mice via cerulein (CAE) injection, alcohol and palmitoleic acid (EtOH + POA) injection, and alcohol diet feeding and cerulein (EtOH + CAE) injection. Primary pancreatic cells and splenocytes were prepared, and multi-dimensional flow cytometry was performed and analyzed by manual gating and computerized PhenoGraph, followed by visualization with t-distributed stochastic neighbor embedding (t-SNE).ResultsCAE treatment induced a time-dependent increase of major innate immune cells and a decrease of follicular B cells, and TCD4+ cells and the subtypes in the pancreas, whereas elicited a reversed pattern in the spleen. EtOH + POA treatment resulted in weaker effects than CAE treatment. EtOH feeding enhanced CAE-induced amylase secretion, but unexpectedly attenuated CAE-induced immune cell regulation. In comparison with manual gating analysis, computerized analysis demonstrated a remarkable time efficiency and reproducibility on the innate immune cells and B cells.ConclusionsThe reverse pattern of increased innate and decreased adaptive immune cells was consistent in the pancreas in CAE and EtOH + POA treatments. Alcohol feeding opposed the CAE effect on immune cell regulation. Together, the immune profiling approach utilized in this study provides a better understanding of overall immune responses in AP, which may facilitate the identification of intervention windows and new therapeutic strategies. Computerized analysis is superior to manual gating by dramatically reducing analysis time.