Cell-mediated Research Articles

Serum Soluble Fibrinogen-Like Protein 2 Estimates Increased T-Helper 2 Cytokine, Elevated Disease Severity, and Unfavorable Outcomes in Allergic Rhinitis Patients.

Soluble fibrinogen-like protein 2 (sFGL2) may be involved in the pathology and progression of allergic rhinitis (AR) through regulating T-helper (Th)2 cell response. This study aimed to explore the ability of sFGL2 to estimate outcomes in AR patients. sFGL2 was detected in the serum sample of 119 AR patients at baseline and 20 healthy controls (HCs) after enrollment by enzyme-linked immunosorbent assay. In AR patients, disease severity was assessed at baseline as well as 4 weeks (W4) and 8 weeks (W8) after enrollment by the total nasal symptom score (TNSS). sFGL2 was higher in AR patients than HCs [median (interquartile range): 100.0 (70.0-154.0) vs 47.0 (31.5-78.5) ng/mL] (P < .001). In AR patients, sFGL2 was positively correlated with TNSS score (P = .002), itching score (P = .037), sneezing score (P = .012), and interleukin (IL)-4 (P = .006) at baseline. TNSS score declined from baseline to W8 in AR patients (P < .001). To further explore the association between sFGL2 levels and treatment outcomes, we compared the sFGL2 levels between patients with TNSS scores at W4 or W8 >4 and those with TNSS scores at W4 or W8 ≤4. It was found that sFGL2 was only increased in AR patients with a TNSS score at W4 >4 versus those with a TNSS score at W4 ≤4 (P = .012). Serum sFGL2 is increased in AR patients, and its high level estimates an increased Th2 cytokine and disease severity as well as poor outcomes.

Myeloid-derived suppressor cells inhibit responses of T follicular helper cells during experimental Plasmodium yoelii infection.

Malaria remains a significant global public health problem. T follicular helper (Tfh) cells, a subset of CD4+ T cells, have the capacity to regulate B cells, plasma cells, and antibody production, among other functions. Myeloid-derived suppressor cells (MDSCs) possess strong immunosuppressive abilities and can negatively regulate various immune responses. However, the role of MDSCs in inhibiting Tfh-cell responses during Plasmodium infection remains unclear. In this study, we investigated the regulatory effect of MDSCs on Tfh cell-mediated immune responses upon Plasmodium infection. We found that the numbers of MDSCs increased upon Plasmodium infection. Further mechanism study revealed that MDSC-derived Arg-1 and PD-L1 prevented Tfh cell proliferation and activation. Conversely, the addition of nor-NOHA or anti-PD-L1 monoclonal antibodies enhanced the proliferation and activation of Tfh cells, indicating that the inhibitory effect of MDSCs on Tfh cells was dependent on Arg-1 and PD-1/PD-L1. Invivo depletion of MDSCs enhanced Tfh-cell responses and antibody production, as well as relieved symptoms of infected mice and improved their survival rates. These findings provide insights into the immunosuppressive role of MDSCs in inhibiting Tfh cell immune responses and further impairing humoral immunity. Our study provides new strategies for malaria prevention and control.

Agaricus bisporus mannose-binding protein stimulates the innate immune cells

Purpose: A lectin-like protein from the mushroom Agaricus bisporus has been shown to slightly increase the proliferation of RAW 264.7 cells. Following its identification as a mannose-binding lectin, henceforth called A. bisporus mannose-binding protein (Abmb), the protein is hypothesized to stimulate the innate immune cells response. The present work was aimed to substantiate that hypothesis. Furthermore, this study complements Abmb exploration as a potential agent for anti-breast cancer, which its treatment is hampered with compromised immunity of patient receiving chemotherapy. Method: Abmb’s effect on the phagocytic activity of the macrophage was measured with FACS. Nitric oxide production was checked using Griess test while expression of the cytokines in the RAW 264.7 cells was analysed at gene and protein level using PCR and FACS, respectively. Abmb’s effect on the expression of surface markers of the human immune cells in the peripheral blood mononuclear cells was checked with specific antibodies for targeted cluster differentiation and analysed using FACS. Result: Abmb increased the phagocytic activity of the macrophage and NO production. Abmb increased the expression of cytokines i.e. tumour necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10. With the peripheral blood mononuclear cells, Abmb activated dendritic and Natural Killer cells, but not the B- or T-cells. Conclusion: Abmb increased the activity of the macrophage cells and activated the immune cells that are related to the innate immune system, particularly the cellular immunity.

Engineering a Near-Infrared Spiro-Based Aggregation-Induced Emission Luminogen for DNAzyme-Sensitized Photothermal Therapy with High Efficiency and Accuracy.

Aggregation-induced emission luminogen (AIEgens)-based photothermal therapy (PTT) has grown into a sparkling frontier for tumor ablation. However, challenges remain due to the uncoordinated photoluminescence (PL) and photothermal properties of classical AIEgens, along with hyperthermia-induced antiapoptotic responses in tumor cells, hindering satisfactory therapeutic outcomes. Herein, a near-infrared (NIR) spiro-AIEgen TTQ-SA was designed for boosted PTT by auxiliary DNAzyme-regulated tumor cell sensitization. TTQ-SA with a unique molecular structure and packing mode was initially fabricated, endowing it with a strong AIE effect, favorable PL quantum yield, and good photothermal performance. DNAzyme, as a gene silencing tool, could alleviate antiapoptosis response during PTT. By integrating TTQ-SA and DNAzyme into folate-modified poly(lactic-co-glycolic acid) (PLGA) polymer, the as-fabricated nanosystem could promote cell apoptosis and sensitize tumor cells to PTT, thereby maximizing the therapeutic outcomes. With the combination of spiro-AIEgen-based PTT and DNAzyme-based gene silencing, the as-designed nanosystem showed promising NIR and photothermal imaging abilities for tumor targeting and demonstrated significant cell apoptotic, antitumor, and antimetastasis effects against orthotopic breast cancer. Furthermore, a synergistic antitumor effect was realized in spontaneous MMTV-PyMT transgenic mice. These findings offer new insights into AIEgen-based photothermal theranostics and DNAzyme-regulated tumor cell sensitization, paving the way for synergistic gene silencing-PTT nanoplatforms in clinical research.

Discovery and engineering of the antibody response to a prominent skin commensal.

The ubiquitous skin colonist Staphylococcus epidermidis elicits a CD8+ T cell response pre-emptively, in the absence of an infection1. However, the scope and purpose of this anti-commensal immune program are not well defined, limiting our ability to harness it therapeutically. Here, we show that this colonist also induces a potent, durable, and specific antibody response that is conserved in humans and non-human primates. A series of S. epidermidis cell-wall mutants revealed that the cell surface protein Aap is a predominant target. By colonizing mice with a strain of S. epidermidis in which the parallel β-helix domain of Aap is replaced by tetanus toxin fragment C, we elicit a potent neutralizing antibody response that protects mice against a lethal challenge. A similar strain of S. epidermidis expressing an Aap-SpyCatcher chimera can be conjugated with recombinant immunogens; the resulting labeled commensal elicits high antibody titers under conditions of physiologic colonization, including a robust IgA response in the nasal and pulmonary mucosa. Thus, immunity to a common skin colonist involves a coordinated T and B cell response, the latter of which can be redirected against pathogens as a novel form of topical vaccination.

Cell-Cultured Influenza Vaccine Enhances IFN-γ+ T Cell and Memory T Cell Responses Following A/Victoria/2570/2019 IVR-215 (A/H1N1) Infection

Background: Influenza remains a significant public health challenge, with vaccination being a substantial way to prevent it. Cell-cultured influenza vaccines have emerged to improve on the drawbacks of egg-based vaccines, but there are few studies focusing on T cell immunity with both types of vaccines. Therefore, we studied the following 2022–2023 seasonal influenza vaccines manufactured and marketed in South Korea with a standard and high dose: cell-based (C_sd and C_hd) and egg-based (E_sd and E_hd) vaccines. Methods: Along with a saline control group, C_sd, C_hd, E_sd, and E_hd vaccines were administered to BALB/c mice, followed by a challenge with the A/Victoria/2570/2019 (H1N1) strain. Results: After the challenge, four out of five mice in the saline group died by day 7 post-infection (P.I.). None of the vaccinated groups experienced over 20% weight loss or any deaths. On day 7 P.I., the lung viral load in the saline group (mean log value of 4.17) was higher than that in the vaccinated groups, with the C_sd group showing the lowest viral load (mean log value of 3.47). The C_sd group showed a significantly high response in macrophage 1 (M1), IFN-γ+ T cells, and tissue-resident memory (TRM) T cells compared with the E_sd group on day 2 P.I. These M1, IFN-γ+ T cells, and TRM cells showed similar trends (p &lt; 0.01). In terms of humoral immunity, only the E_hd group showed HAI titers above 40 for all four strains before and after the challenge. Conclusions: The high levels of T cells in the cell-cultured vaccines suggest, pending further real-world research, that these vaccines may offer advantages.

Unraveling the impact of SARS-CoV-2 mutations on immunity: insights from innate immune recognition to antibody and T cell responses

Throughout the COVID-19 pandemic, the emergence of new viral variants has challenged public health efforts, often evading antibody responses generated by infections and vaccinations. This immune escape has led to waves of breakthrough infections, raising questions about the efficacy and durability of immune protection. Here we focus on the impact of SARS-CoV-2 Delta and Omicron spike mutations on ACE-2 receptor binding, protein stability, and immune response evasion. Delta and Omicron variants had 3–5 times higher binding affinities to ACE-2 than the ancestral strain (KDwt = 23.4 nM, KDDelta = 8.08 nM, KDBA.1 = 4.77 nM, KDBA.2 = 4.47 nM). The pattern recognition molecule mannose-binding lectin (MBL) has been shown to recognize the spike protein. Here we found that MBL binding remained largely unchanged across the variants, even after introducing mutations at single glycan sites. Although MBL binding decreased post-vaccination, it increased by 2.6-fold upon IgG depletion, suggesting a compensatory or redundant role in immune recognition. Notably, we identified two glycan sites (N717 and N801) as potentially essential for the structural integrity of the spike protein. We also evaluated the antibody and T cell responses. Neutralization by serum immunoglobulins was predominantly mediated by IgG rather than IgA and was markedly impaired against the Delta (5.8-fold decrease) and Omicron variants BA.1 (17.4-fold) and BA.2 (14.2-fold). T cell responses, initially conserved, waned rapidly within 3 months post-Omicron infection. Our data suggests that immune imprinting may have hindered antibody and T cell responses toward the variants. Overall, despite decreased antibody neutralization, MBL recognition and T cell responses were generally unaffected by the variants. These findings extend our understanding of the complex interplay between viral adaptation and immune response, underscoring the importance of considering MBL interactions, immune imprinting, and viral evolution dynamics in developing new vaccine and treatment strategies.

The effect of sleep and shift work on the primary immune response to messenger RNA-based COVID-19 vaccination.

Shift work can cause circadian misalignment, which often results in sleeping problems and has been associated with immune dysfunction. To better understand the impact of shift work on a primary immune response to vaccination, we compared severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-specific humoral and cellular immune responses after one injection of the messenger RNA (mRNA)-1273 vaccine between day workers (n = 24) and night shift workers (n = 21). In addition, duration and quality of sleep were assessed for a period of 7 days around the time of vaccination using actigraphy and daily sleep diaries, and their relationship with immunogenicity of mRNA-1273 vaccination was studied. We found that median total sleep time on the 2 days immediately after vaccination, which coincided with the days that night shift workers worked night shifts, was significantly lower in night shift workers (342 and 318 min) than day workers (431 and 415 min) (both p < 0.001). There was no difference in sleep quality between day workers and night shift workers. Furthermore, no difference in the antibody response between the two groups was observed, yet night shift workers had a significantly higher virus-specific T-cell response than day workers 28 days after immunisation (p = 0.013). Multivariate regression analysis showed no association between sleep duration, sleep quality and SARS-CoV-2-specific humoral or cellular immune responses. Collectively, these findings indicate that shift work-induced sleep loss and night shift work have little to no effect on the primary immune response to mRNA-based COVID-19 vaccination.

Impact of administration routes and dose frequency on the toxicology of SARS-CoV-2 mRNA vaccines in mice model.

The increasing use of SARS-CoV-2 mRNA vaccines has raised concerns about their potential toxicological effects, necessitating further investigation to ensure their safety. To address this issue, we aimed to evaluate the toxicological effects of SARS-CoV-2 mRNA vaccine candidates formulated with four different types of lipid nanoparticles in ICR mice, focusing on repeated doses and administration routes. We conducted an extensive analysis in which mice received the mRNA vaccine candidates intramuscularly (50 μg/head) twice at 2-week intervals, followed by necropsy at 2 and 14 dpsi (days post-secondary injection). In addition, we performed a repeated dose toxicity test involving three, four, or five doses and compared the toxicological outcomes between intravenous and intramuscular routes. Our findings revealed that all vaccine candidates significantly induced SARS-CoV-2 spike protein-specific IgG and T cell responses. However, at 2 dpsi, there was a notable temporary decrease in lymphocyte and reticulocyte counts, anemia-related parameters, and significant increases in cardiac damage markers, troponin-I and NT-proBNP. Histopathological analysis revealed severe inflammation and necrosis at the injection site, decreased erythroid cells in bone marrow, cortical atrophy of the thymus, and increased spleen cellularity. While most toxicological changes observed at 2 dpsi had resolved by 14 dpsi, spleen enlargement and injection site damage persisted. Furthermore, repeated doses led to the accumulation of toxicity, and different administration routes resulted in distinct toxicological phenotypes. These findings highlight the potential toxicological risks associated with mRNA vaccines, emphasizing the necessity to carefully consider administration routes and dosage regimens in vaccine safety evaluations, particularly given the presence of bone marrow and immune organ toxicity, which, though eventually reversible, remains a serious concern.

Insights into the paradoxical effect of smoking on vasculitis: a comprehensive review.

Vasculitis is a group of uncommon diseases characterized by inflammation of blood vessels, which contributes to the organ ischemia and damage. Cigarette smoke contains a high concentration of various toxins, which have the potential to affect the immune response and development of autoimmune/autoinflammatory rheumatic diseases including vasculitis. Smoking influences both innate and adaptive immune systems and plays binary functions in modulating immunity by either aggravating pathogenic immune responses or attenuating defensive immunity. Smoking contributes to the pathogenesis of autoimmune diseases by various mechanisms including induction of tissue damage and apoptosis, changes in innate immune function and production of proinflammatory cytokines, changes in humoral immunity and T cell responses and anti-estrogen effects. In this review, we considered the available evidence on the association between smoking with the risk, clinical manifestations, response to treatment and outcomes of vasculitis, and the effect of smoking cessation on these parameters. In conclusion, despite inconclusive evidence of an increased risk of giant cell arthritis and anti-neutrophil cytoplasmic autoantibody associated vasculitis (AAV) in smokers, there is strong evidence that smokers have a lower risk of Behcet's disease (BD). Furthermore, smoking changes the clinical picture and outcomes of BD and AAV.

Effects of Ca2+ on the Structure and Dynamics of PIP3 in Model Membranes Containing PC and PS.

Phosphatidylinositol phosphates (PIPs) are a family of seven different eukaryotic membrane lipids that have a large role in cell viability, despite their minor concentration in eukaryotic cellular membranes. PIPs tightly regulate cellular processes, such as cellular growth, metabolism, immunity, and development through direct interactions with partner proteins. Understanding the biophysical properties of PIPs in the complex membrane environment is important to understand how PIPs selectively regulate a partner protein. Here, we investigate the structure and dynamics of PIP3 in lipid bilayers that are simplified models of the natural membrane environment. We probe the effects of the anionic lipid phosphatidylserine (PS) and the divalent cation Ca2+ by using full-length lipids in well-formed bilayers. We used solution and solid-state NMR on naturally abundant 1H, 31P, and 13C atoms combined with molecular dynamics (MD) simulations to characterize the structure and dynamics of PIPs. 1H and 31P 1D spectra show good resolution at temperatures above the phase transition with isolated peaks in the headgroup, interfacial, and bilayer regions. Site-specific assignment of the chemical shifts of these reporters enables the measurement of the effects of Ca2+ and PS at the single atom level. In particular, the resolved 31P signals of the PIP3 headgroup allow for extremely well-localized information about PIP3 phosphate dynamics, which the MD simulations can further explain. A quantitative assessment of cross-polarization kinetics provides additional dynamics measurements for the PIP3 headgroups.

Elemental composition of sputum, immune status, and features of pharyngeal microbiota in workers of the anode department of a copper smelter

The development of bronchial and pulmonary pathology depends on characteristics of the aerosols and individual resistance of the body, largely determined by the immune system, to their exposure. Mucous membranes of a healthy person are colonized by saprophytic bacteria or normal microbiota. Yet, changes in the composition and properties of the microbial landscape induced by adverse environmental factors increase the risk of inflammation and contribute to changes in the reactivity of the immune system. The study aims to establish the immune status, microbiota of the pharyngeal mucosa, and the elemental composition of sputum, given its cellular composition, in workers of the anode department of a copper smelter. We tested sputum culture and elemental composition, established clinical and immunological status, and analyzed pharyngeal microbiota in 28 workers of the anode department of a copper smelter. Based on the results of testing, the subjects were divided into two groups with (1) and without (2) leukocytosis (&gt;10 cells in the field of view) detected in sputum. We found no acute or chronic diseases of the bronchi and lungs in the examined workers. In group 1, we observed shifts in the immune response manifested by a decrease in the number of immunocompetent cells, activation of neutrophil phagocytosis, formation of reactive oxygen species in neutrophils, and displacement of normal microbiota of the oropharyngeal mucosa by opportunistic microorganisms (in 100% versus 35%, p&lt;0.05). The elemental composition of sputum was noted for a higher frequency of iron and lead detection and increased concentrations of barium, lead, iron, arsenic, molybdenum, antimony, vanadium, and cadmium. We established a significant effect of metals contained in sputum on cellular, humoral, and local immunity. The inflammatory bronchopulmonary response to toxic aerosol exposure is accompanied by changes in the cellular and phagocytic links of immunity, as well as in the microbial landscape on the mucous membrane of the upper respiratory tract characterized by displacement of normal biota by opportunistic microorganisms. Limitations. This study has certain limitations associated with the small sample size and the lack of opportunity to compare metal concentrations found in sputum with reference levels. Ethics. The study was approved by the Local Ethics Committee of the Yekaterinburg Medical Research Center for Prophylaxis and Health Protection in Industrial Workers (extract from the minutes of the meeting No. 8 dated 12/26/2016).

Monocyte uptake of polymeric peptidoglycan is bimodal and governed by complement C3 and C4 opsonins.

Peptidoglycans (PGNs) are structural polymers of the bacterial cell wall and a common microbial molecular pattern encountered by our immune system daily. Low levels of PGNs are constitutively present in the systemic circulation in humans and elevate during inflammatory pathologies. Since all known PGN sensors are intracellular, PGN internalization is a prerequisite for the initiation of cellular immune responses. Here we report the mechanisms controlling the recognition and uptake of polymeric PGNs by circulating human mononuclear phagocytes. We found that complement C3 and C4 opsonins govern PGN recognition and internalization, but no single opsonin is indispensable due to multiple uptake redundancies. We observed a bimodal internalization of polymeric PGNs with distinct requirements for complement C4. At low PGN concentrations, C3 mediated PGN recognition by surface receptors while the efficient internalization of PGN polymers critically required C4. Supraphysiologic PGN concentrations triggered a secondary uptake modality that was insensitive to C4 and mediated instead by C3 engagement of complement receptors 1 and 3. To our knowledge this is the first description of non-overlapping C3 and C4 opsonophagocytoses working in parallel. Controlling these uptake mechanisms has the potential to modulate PGN clearance and/or the dysregulated immune responses during bacterial infections.

Dysfunction and regulatory interplay of T and B cells in chronic hepatitis B: immunotherapy and emerging antiviral strategies

In the context of chronic hepatitis B virus (HBV) infection, the continuous replication of HBV within host hepatocytes is a characteristic feature. Rather than directly causing hepatocyte destruction, this replication leads to immune dysfunction and establishes a state of T-B immune tolerance. Successful clearance of the HBV virus is dependent on the close collaboration between humoral and cellular immunity. Humoral immunity, mediated by B-cell subpopulations, and cellular immunity, dominated by T-cell subpopulations show varying degrees of dysfunction during chronic hepatitis B (CHB). Notably, not all T- and B-cells produce positive immune responses. This review examine the most recent developments in the mutual regulation of T-B cells during chronic HBV infection. Our focus is on the prevailing immunotherapeutic strategies, such as T cell engineering, HBV-related vaccines, PD-1 inhibitors, and Toll-like receptor agonists. While nucleos(t)ide analogues (NUCs) and interferons have notable limitations, including inadequate viral suppression, drug resistance, and adverse reactions, several HBV entry inhibitors have shown promising clinical efficacy. To overcome the challenges posed by NUCs or monotherapy, the combination of immunotherapy and novel antiviral agents presents a promising avenue for future CHB treatment and potential cure.

Booster COVID-19 mRNA vaccination ameliorates impaired B-cell but not T-cell responses in older adults

Age-associated differences in the effect of repetitive vaccination, particularly on memory T-cell and B-cell responses, remain unclear. While older adults (aged ≥65 years) exhibited enhanced IgG responses following COVID-19 mRNA booster vaccination, they produced fewer spike-specific circulating follicular helper T cells-1 than younger adults. Similarly, the cytotoxic CD8+ T-cell response remained diminished with reduced PD-1 expression even after booster vaccination compared with that in younger adults, suggesting impaired memory T-cell activation in older adults. In contrast, although B-cell responses in older adults were weaker than those in younger adults in the primary response, the responses were significantly enhanced upon booster vaccination, reaching levels comparable with that observed in younger adults. Therefore, while booster vaccination ameliorates impaired humoral immunity in older adults by efficiently stimulating memory B-cell responses, it may less effectively enhance T-cell-mediated cellular immunity. Our study provides insights for the development of effective therapeutic and vaccine strategies for the most vulnerable older population.

Development and application of an uncapped mRNA platform

Background A novel uncapped mRNA platform was developed. Methods Five lipid nanoparticle (LNP)-encapsulated mRNA constructs were made to evaluate several aspects of our platform, including transfection efficiency and durability in vitro and in vivo and the activation of humoral and cellular immunity in several animal models. The constructs were eGFP-mRNA-LNP (for enhanced green fluorescence mRNA), Fluc-mRNA-LNP (for firefly luciferase mRNA), SδT-mRNA-LNP (for Delta strain SARS-CoV-2 spike protein trimer mRNA), gDED-mRNA-LNP (for truncated glycoprotein D mRNA coding ectodomain from herpes simplex virus type 2 (HSV2)) and gDFR-mRNA-LNP (for truncated HSV2 glycoprotein D mRNA coding amino acids 1–400). Results Quantifiable target protein expression was achieved in vitro and in vivo with eGFP- and Fluc-mRNA-LNP. SδT-mRNA-LNP, gDED-mRNA-LNP and gDFR-mRNA-LNP induced both humoral and cellular immune responses comparable to those obtained by previously reported capped mRNA-LNP constructs. Notably, SδT-mRNA-LNP elicited neutralizing antibodies in hamsters against the Omicron and Delta strains. Additionally, gDED-mRNA-LNP and gDFR-mRNA-LNP induced potent neutralizing antibodies in rabbits and mice. The mRNA constructs with uridine triphosphate (UTP) outperformed those with N1-methylpseudouridine triphosphate (N1mψTP) in the induction of antibodies via SδT-mRNA-LNP. Conclusions Our uncapped, process-simplified and economical mRNA platform may have broad utility in vaccines and protein replacement drugs. KEY MESSAGES The mRNA platform described in our paper uses internal ribosome entry site (IRES) (Rapid, Amplified, Capless and Economical, RACE; Register as BH-RACE platform) instead of caps and uridine triphosphate (UTP) instead of N1-methylpseudouridine triphosphate (N1mψTP) to synthesize mRNA. Through the self-developed packaging instrument and lipid nanoparticle (LNP) delivery system, mRNA can be expressed in cells more efficiently, quickly and economically. Particularly exciting is that potent neutralizing antibodies against Delta and Omicron real viruses were induced with the new coronavirus S protein mRNA vaccine from the BH-RACE platform.

Adenosine Uptake through the Nucleoside Transporter ENT1 Suppresses Antitumor Immunity and T Cell Pyrimidine Synthesis.

Immunosuppression by adenosine is an important cancer immune checkpoint. Extracellular adenosine signals through specific receptors and can be transported across the cell membrane through nucleoside transporters. While adenosine receptors are well-known to regulate tumor immunity, the impact of adenosine transporters remains unexplored. In this study, we investigated the effect on tumor immunity of equilibrative nucleoside transporter-1 (ENT1), the major regulator of extracellular adenosine concentrations. Blocking or deleting host ENT1 significantly enhanced CD8+ T cell-dependent antitumor responses. Tumors inoculated into ENT1-deficient mice showed increased infiltration of effector CD8+ T cells with an enhanced cytotoxic transcriptomic profile and significant upregulation of granzyme B, IFN-γ, IL-2, TNF-α, and CXCL10. ENT1-deficiency was further associated with decreased tumor-infiltrating T regulatory cells and CD206high macrophages and suppressed CCL17 production. ENT1-deficiency notably potentiated the therapeutic activity of PD-1 blockade. T cells upregulated ENT1 upon activation, and blocking ENT1 enhanced their function when co-cultured with cognate antigen/HLA-matched melanoma cells. Mechanistically, ENT1-mediated adenosine uptake inhibited the activity of phosphoribosyl pyrophosphate synthetase (PRPS) in activated T cells, thereby suppressing production of uridine 5'-monophosphate (UMP) and its derivatives required for DNA and RNA synthesis. In summary, this study identified ENT1-mediated adenosine uptake as an important mechanism of adenosine-mediated immunosuppression and pyrimidine starvation that can be targeted to enhance antitumor T cell responses.

Transcriptome Analysis Reveals Sertoli Cells Adapting Through Redox and Metabolic Pathways Under Heat Stress in Goats

Background/Objectives: Climate change-induced temperature elevations pose significant challenges to livestock reproduction, particularly affecting testicular function in small ruminants. This study investigates the acute heat-stress response in goat Sertoli cells (SCs), aiming to elucidate the molecular mechanisms underlying heat-induced damage to male reproductive tissues. Methods: SCs were isolated from testes of 4-month-old black goats and exposed to heat stress (44 °C for 2.5 h). We employed transcriptome sequencing, CCK-8 assay, electron microscopy, ROS measurement, autophagy detection, Western blot analysis, and lactate concentration measurement. Bioinformatics analyses including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and protein–protein interaction network analyses were performed on the transcriptome data. Results: Heat stress significantly reduced SC viability, induced oxidative stress and autophagy, and altered gene expression profiles. We identified 1231 significantly differentially expressed genes, with significant enrichment in membrane-related processes and metabolic pathways. Metabolism-related genes, including PKLR, ACOT11, and LPCT12, were significantly downregulated. Protein–protein interaction network analysis revealed ten hub genes potentially crucial in the heat-stress response: HSP90AA1, HSPA5, BAG3, IGF1, HSPH1, IL1A, CCL2, CXCL10, ALB, and CALML4. Conclusions: This study provides comprehensive insights into the molecular mechanisms underlying goat SC response to heat stress. The identified genes and pathways, particularly those related to metabolism and stress response, offer potential targets for developing strategies to mitigate heat-stress effects on livestock reproduction. These findings contribute to our understanding of climate change impacts on animal husbandry and may inform the development of heat-stress resistant livestock lines.

Evolution of the ocular immune system.

The evolution of the ocular immune system should be viewed within the context of the evolution of the immune system, and indeed organisms, as a whole. Since the earliest time, the most primitive responses of single cell organisms involved molecules such as anti-microbial peptides and behaviours such as phagocytosis. Innate immunity took shape ~2.5 billion years ago while adaptive immunity and antigen specificity appeared with vertebrate evolution ~ 500 million years ago. The invention of the microscope and the germ theory of disease precipitated debate on cellular versus humoral immunity, resolved by the discovery of B and T cells. Most recently, our understanding of the microbiome and consideration of the host existing symbiotically with trillions of microbial genes (the holobiont), suggests that the immune system is a sensor of homoeostasis rather than simply a responder to pathogens. Each tissue type in multicellular organisms, such as vertebrates, has a customised response to immune challenge, with powerful reactions most evident in barrier tissues such as the skin and gut mucosa, while the eye and brain occupy the opposite extreme where responses are attenuated. The experimental background which historically led to the concept of immune privilege is discussed in this review; however, we propose that the ocular immune response should not be viewed as unique but simply an example of how the tissues variably respond in nature, more or less to the same challenge (or danger).

Identification and functional analysis of novel biomarkers related to retinal neovascularization in proliferative diabetic retinopathy

Background Proliferative diabetes retinopathy (PDR) seriously affects the vision of patients. Exploring the key genes of retinal neovascularization is crucial for developing new biomarkers and therapeutic targets. Objective This study aimed to identify key genes associated with retinal neovascularization in Proliferative Diabetic Retinopathy (PDR), intending to develop new biomarkers and therapeutic targets. This would further our understanding of the progression of diabetic retinopathy and improve patient prognosis. Methods The gene data from 36 diabetic retinopathy patient samples and 45 samples from healthy volunteers or diabetic patients were selected from the GEO DataSets (Gene Expression Omnibus), specifically datasets GSE102485 and GSE160310. Utilizing the SVA algorithm to merge datasets and the limma package in R to identify differentially expressed genes (DEGs), we conducted a bioinformatic analysis of diabetic retinopathy. Functional insights were gained through DAVID database analyses, while STRING database-derived Protein-Protein Interaction (PPI) networks visualized in Cytoscape provided further context. Key genes were identified through LASSO regression and SVM analyses, with ROC curves assessing their diagnostic value. Single gene set enrichment analysis (GSEA) enhanced our understanding of the perturbed biological processes and pathways, advancing knowledge of diabetic retinopathy at the genomic level. Results A rigorous bioinformatic analysis yielded a comprehensive list of 1139 differentially expressed genes (DEGs), of which six pivotal genes—KDM5D, AC007040.11, AC015688.3, NLRP2, GYPC, and TMSB4Y—were identified as central to the study. These six genes consistently demonstrated a high diagnostic accuracy, with each exhibiting an area under the receiver operating characteristic (ROC) curve (AUC) exceeding 0.75. Gene Ontology (GO) enrichment analysis elucidated their primary roles in intricate biological processes, including inflammatory and immune responses, T-cell activation, cell apoptosis, and angiogenesis. Moreover, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment revealed their involvement in crucial signaling cascades such as cytokine-cytokine receptor interactions, cell adhesion molecule pathways, PI3K-Akt signaling, and hematopoietic cell lineage, further substantiating their significance in the pathogenesis of diabetic retinopathy. Conclusions In this research, we conducted a comprehensive gene expression analysis in patients with and without PDR, identifying differentially expressed genes, pivotal biomarkers, and critical pathways potentially involved in PDR progression. Our findings enhance understanding of PDR's molecular dynamics and offer potential avenues for improved prognostication and therapeutic intervention, despite limitations due to sample group heterogeneity.