Immune Stimulation Research Articles

An Overview on Amaranthus cruentus L.: Alternative Source of Crop, its Economical Uses and Associated Beneficial Microbes

Amaranthus cruentus L.is a medicinally important locally available tropical plant belonging to the family Amaranthaceae. In present scenario, this plant is designated as ‘alternative source of crop’ or ‘Pseudocereal’ having various nutritional properties (Protein, carbohydrate, fatty acid, Vitamins, Minerals). Moreover, this plant has various medicinal properties also i.e. antioxidant, antihypertensive, antidiabetic, immune stimulation, anthelmintic, hepatoprotective, antimalarial etc. This plant has Worldwide distribution and short growing period of time. It is also regarded as Fodder crop and other industrial uses. The microbes associated with this plant help to protect the host plants from both pathogenic microorganisms and adverse climatic conditions by secreting different types of bioactive secondary metabolites i.e. phenolics, flavonoids, terpenoids, terpenes, tannins, alkaloids, quinones, anthocyanin, saponin, steroids and promote growth and development of the plant. Some unique features of this plant are also descried here. This review intends to expand the understanding of taxonomic characteristics of this plant, economic uses, associated beneficial microbes and their application for sustainability of this alternative source of crop. The cultivation of A. cruentus possess a good attraction among both the researchers and farmers and cultivation procedure is more easy than other species. Some species of this genus are not cultivated yet, they are under process. Additionally, this species is more adorably cultivated due to the high nutritional value of its seeds.

Engineered Immunomodulatory Extracellular Vesicles from Epithelial Cells with the Capacity for Stimulation of Innate and Adaptive Immunity in Cancer and Autoimmunity.

Extracellular vesicles (EVs) are generated in all cells. Systemic administration of allogenic EVs derived from epithelial and mesenchymal cells has been shown to be safe, despite carrying an array of functional molecules, including thousands of proteins. To address whether epithelial cell-derived EVs can be modified to acquire the capacity to induce an immune response, we engineered 293T EVs to harbor the immunomodulatory molecules CD80, OX40L, and PD-L1. We demonstrated abundant levels of these proteins in the engineered cells and EVs. Functionally, the engineered EVs efficiently elicited positive and negative costimulation of human and murine T cells. In the setting of cancer and autoimmune hepatitis, the engineered EVs modulated T cell functions and altered disease progression. OX40L EVs also provided enhanced antitumor activity in combination with anti-CTLA-4 in melanoma-bearing mice. In addition, we added multiple immunomodulatory proteins in EVs (EVmIM), attempting to elicit an immune response in both lymphoid and myeloid compartments. The EVmIM containing CD80, 4-1BBL, CD40L, CD2, and CD32 engaged both T cells and antigen presenting cells (APCs) in melanoma tumors, demonstrating the capacity for EVmIM to elicit antitumor activity. Our work provides evidence that EVs can be engineered to induce specific immune responses with translational potential to modulate immune cell functions in pathological settings.

Toripalimab: A new age in fighting Nasopharyngeal Carcinoma

Madam, Nasopharyngeal carcinomas (NPCs) lie amongst a group of malignant tumours whose incidence has steadily increased over the past 30 years.[1] These have a significantly higher incidence rate in Asian populations, which account for over 70% of new cases worldwide.[2] The leading risk factors associated with NPCs are tobacco and alcohol consumption, followed by HPV infections. Usually diagnosed at advanced stages, a combination of chemo and radiotherapy is the standard treatment. However, they have one of the lowest survival rates compared to other cancers.[1] Programmed cell death protein 1(PD - 1) is a receptor that is expressed on T cells, B cells, macrophages, dendritic cells and natural killer cells. Binding of PD-1 with its ligands, namely PD-L1 and PD-L2, transmits an inhibitory signal, which decreases the proliferation and production of cytokines by T cells, thereby inhibiting immune system stimulation. Toripalimab is a monoclonal antibody that blocks interactions between the PD - 1 and PD-L1 or PD-L2.[3] Toripalimab was approved by the FDA on 10/27/2023 with the following indications; first line treatment of locally advanced, recurrent or metastatic NPC(RM-NPC) in combination with standard chemotherapy using cisplatin plus gemcitabine, and as a lone agent in patients with RM-NPC showing disease progression despite being on or after platinum containing chemotherapy.[3] The approval was based off of two clinical trials; POLARIS-02 and JUPITER-02,[4][5] which suggested that toripalimab combined with standard chemotherapy provided statistically as well as clinically significant, progression free survival and overall survival benefits compared to standard chemotherapy alone. The aforementioned clinical trials showed a manageable safety profile with the overall rate of grade 3 or greater side effects being similar in the two groups- toripalimab plus standard chemotherapy versus standard chemotherapy alone and that the most common side effects were mainly due to standard chemotherapy itself. The main immune related adverse effects reported were hypothyroidism, hyperthyroidism, abnormal liver function, interstitial lung disease and dermatomyositis.[4][5] In recent years immunotherapy has emerged as a highly effective avenue when used in combination with targeted therapies. This combination enhances the immune cells’ attack capabilities and thereby enhances the efficacy of the treatment.[3] Toripalimab is the first FDA- approved immune checkpoint mediator for the treatment of RM-NPC and the promising results from the clinical trials alongside the tractable safety profile support the use of Toripalimab in combination with gemcitabine-cisplatin chemotherapy as the new standard of care for this population.

Prenatal Lipopolysaccharide Stimulation Modulates Gastrointestinal Immunity and Oxidative Status in Weaned Pigs.

Gastrointestinal immunity and antioxidant defenses may be bolstered in young animals through prenatal immune system stimulation (PIS), but this is largely uninvestigated in swine. This study tested the hypothesis that PIS could regulate offspring's gastrointestinal immune response and oxidative stress profile. To this end, a PIS model was utilized in sows, delivering low-dose LPS during the final third of gestation to target the developing immune system. On day 78±1.8 of gestation, 14 Camborough sows (parity = 2.6±1.4) received either saline (CON) or LPS from E. coli O111:B4 (2.5 μg/kg of body weight). A subset of 34 weaned barrows (n=17 CON, PIS), weaned at 21±1.3 days, were anesthetized for subcutaneous temperature loggers and jugular catheter placement. Following recovery, all pigs received an intravenous injection of LPS (10 μg/kg body weight) from E. coli O111:B4. Our findings demonstrate that PIS enhances the gut immune response by upregulating key inflammatory cytokines, indicative of a proinflammatory profile. Consistently across the jejunum and ileum, SCF was modulated with heightened expression in PIS than CON (P≤ 0.05). In the ileum alone, PIS exhibited heightened expression of proinflammatory cytokines and chemokines, including TNFα, IL-6, IL-1β, and CCL3L1, compared to CON (P≤ 0.05). Exposure to PIS resulted in reduced systemic total antioxidant capacity (TAC) at hours 2-4 post-challenge (P= 0.004). Piglets exposed to PIS had decreased jejunal tissue malondialdehyde (MDA) concentrations (P= 0.049). Together, these data indicate that exposure to PIS alters the inflammatory profile of the GI immune response and oxidative status in weaned pigs.

Bivalent OX40 Aptamer and CpG as Dual Agonists for Cancer Immunotherapy.

Cancer immunotherapy has revolutionized cancer treatment by harnessing the body's immune system to recognize and attack tumors. Over the past 25 years, the use of blocking antibodies has fundamentally transformed the landscape of cancer therapy. However, despite extensive research, agonist antibodies targeting costimulatory receptors such as ICOS, GITR, OX40, CD27, and 4-1BB have consistently underperformed in clinical trials over the past 15 years, failing to meet the anticipated success. One reason the agonist antibodies failed is that researchers escalated the dose to the highest tolerable level, which can lead to cell exhaustion, especially when used as a single agent. In this study, we introduced novel in situ therapeutic agents by combining a bivalent RNA aptamer of OX40, biROX40, which binds to two copies of the OX40 receptor as an agonist, with CpG, a toll-like receptor 9 (TLR9) immune stimulator. These agents were specifically designed for lymphoma treatment, with the dose reduced to the lowest bioactive amount to maximize efficacy while minimizing potential side effects. BiROX40 and CpG exhibited a dual immune activation effect and demonstrated a synergistic response even at extremely low dose of 0.32 mg/kg (5.75 μg per mouse) for biROX40 and moderate dose of 1.39 mg/kg (25 μg per mouse) for CpG, resulting in remarkable antitumor efficacy. This effect was achieved through the promotion of intratumoral CD8+ T cell proliferation and cytokine secretion, inhibition of regulatory T cell (Treg) proliferation, and enhanced generation and proliferation of memory T cells in immune organs. The agonistic effects of these reagents led to tumor regression not only at the treated sites but also at distant, nontreated locations in the animal models. This outcome highlighted the induction of a robust systemic antitumor immune response, which effectively suppressed tumor recurrence. This in situ combination therapy, utilizing low-dose biROX40 alongside CpG, offers a straightforward and widely applicable strategy to enhance immune responses in cancer immunotherapy. This approach overcomes the limitations of high-dose single-agent anti-OX40 therapies (whether antibodies or aptamers), including immune cell exhaustion and diminished efficacy.

A case report on Pseudo Felty’s syndrome

Pseudo-Felty’s syndrome is a rare disease entity defined by the presence of monoclonal expansion of T lymphocytes with neutropenia in association with rheumatoid arthritis. T-large granular lymphocyte (LGL) leukemia is a lymphoproliferative disease with an indolent behavior. LGL leukemia arises in a setting of sustained immune stimulation, such as autoimmune disorders and chronic viral infections, where there is a clonal expansion of T-LGL (cluster of differentiation [CD]3+/CD8+) cells. In this article, we present the case of a 60-year-old gentleman with Pseudo-Felty’s syndrome who presented with LGL proliferation. He is presently stable on disease modifying anti-rheumatoid drugs including methotrexate.

DOP064 Evaluation of pharmacological interventions in human Precision-Cut Intestinal Slices: insights into tissue-resident immunity in Inflammatory Bowel Diseases

Abstract Background Inflammatory Bowel Diseases (IBD), including Crohn's Disease (CD) and Ulcerative Colitis (UC), are marked by dysregulated immune responses and significant T cell infiltration, leading to chronic intestinal inflammation. The global burden of IBD is increasing, yet its pathogenesis remains only partly understood. Our study aims to gain deeper insights into the immunological responsiveness of diseased tissue using ex vivo primary human intestinal tissue slices (Precision Cut Intestinal Slices; PCIS). PCIS are ultrathin tissue slices of the intestine of human donors that remain viable in culture, preserve intestinal architecture, and maintain all relevant intestinal cells including tissue-resident immune cells. Here, we specifically investigated the effects of the JAK1 inhibitor filgotinib and the calcineurin inhibitor pimecrolimus on tissue-resident immune cell responses in PCIS ex vivo. Methods PCIS from ileum and colon resections of Crohn’s disease (CD) and non-CD patients were stimulated ex vivo with Concanavalin A, anti-CD3/CD28 antibodies and IL-1β. Induced responses were modulated with anti-inflammatory treatments such as pimecrolimus and filgotinib for 24h. We evaluated tissue viability (LDH-assay), cytokine release (multiplex ELISA), and PCIS morphology (H&E-staining and Immune fluorescence). Results CD and non-CD intestinal tissue slices maintained viability and intestinal morphology, including disease-specific features in ex vivo culture such as immune cell infiltrates and higher abundance of CD4+ and CD8+ T cells. Ex vivo treatment with various immune stimulations revealed distinct disease-specific cytokine release patterns, with a primary focus on Th1 and Th17 responses, but also including other Th cell, innate and epithelial responses. Filgotinib (1-100 μM) dose-dependently inhibited the secretion of general proinflammatory and T cell-specific cytokines (maximal decrease e.g. IL-8: 8-fold, IL-6: 50-fold, IL-17A: 27-fold). Pimecrolimus (25 µM) specifically decreased T cell-specific cytokines (maximal decrease e.g. IL-2: 13-fold, IL-17A: 22-fold, IL-17F: 27-fold or IFNγ: 7-fold) while the release of general proinflammatory mediators like IL-8 was less affected. Conclusion Our data show the anti-inflammatory effects of filgotinib and pimecrolimus in primary intestinal tissue slices ex vivo. These effects include modulation of various T cell-associated cytokines and other molecules involved in the calcineurin and JAK-STAT pathways. Notably, the different treatments showed distinct effects on cytokine release profiles. This study underscores the potential of PCIS to elucidate the mechanisms of JAK1 inhibitors, advancing IBD treatment strategies and providing a test platform for new drugs in a complex human tissue system.

Molecular responses of Mytilus coruscus hemocytes to lipopolysaccharide and peptidoglycan as revealed by 4D-DIA based quantitative proteomics analysis.

Mytilus are sessile filter feeders that live in close contact with numerous marine microorganisms. Hemocytes, the immunocompetent cells of Mytilus, participate in the immune response in a very efficient manner. Lipopolysaccharide (LPS) and peptidoglycan (PGN) follow specific microbe/pathogen-associated molecular patterns (MAMPs or PAMPs) and are involved in immune stimulation in host cells. This study evaluated the molecular profiles and reactions at protein level of Mytilus hemocytes after stimulation with LPS and PGN. Mytilus coruscus was challenged in vivo with LPS and PGN. The hemocytes were collected after 48 h and analyzed for quantitative proteomics, cell subpopulations, and the free amino acid composition. 4D-DIA technology-based proteomic analysis revealed different protein profiles, as well as different responses at protein level, under either the LPS or PGN challenge. C-type lectins, collagens, and CD151 protein were highly upregulated in LPS-challenged mussels, while phospholipase A2 and dCMP deaminase were highly upregulated in PGN-challenged mussels. Moreover, LPS challenge disrupted dsRNA-mediated translation and stimulated energy-related metabolism, while PGN challenge stimulated proteins involved in the inflammatory response and suppressed amino acid metabolism. In addition, the LPS and PGN challenges differed in their effects on the free amino acid composition and granulocytes ratio of the hemocytes. These findings highlight the different strategies employed by mussel hemocytes in response to different MAMPs, providing insights into the effects of LPS and PGN on Mytilus.

In Vitro and In Vivo Antioxidant and Immune Stimulation Activity of Wheat Product Extracts.

Inflammation and oxidative stress are the main pathogenetic pathways involved in the development of several chronic degenerative diseases. Our study is aimed at assessing the antioxidant and anti-inflammatory activity of hydroalcoholic extracts obtained from wheat and its derivatives. The content of total phenolic and total flavonoid compounds and antioxidant activity were carried out by ABTS and DPPH assays. The ability of wheat extracts to promote microglia polarization towards an anti-inflammatory phenotype was evaluated analyzing the increased expression of anti-inflammatory markers by real-time qPCR and immunofluorescence assays. Antioxidant activity of all the extracts was evaluated in C. elegans by analyzing ROS levels and the expression of the antioxidant enzymes GST-4 and SOD-3 by real-time qPCR and fluorescence experiments. The expression of key genes involved in the innate immune response and stress resistance pathways-daf-16, sek-1, and pmk-1-was evaluated by real-time qPCR. Wheat extracts showed the ability to polarize microglia cells towards an anti-inflammatory phenotype, even after the addition of LPS. An antioxidant response was detected both in microglia and in Caenorhabditis elegans nematode, where the extracts also implemented an anti-stress resilience response and stimulated the innate immunity. The present study shows that wheat seeds, flour, chaff, and pasta present anti-inflammatory as well as antioxidant activities and may be considered as prospective positive health agents for the preparation of functional foods. Moreover, the valorization of by-products from agricultural and agro-industrial activities would also have significant implications in terms of circular economy.

A c-type lectin HcLec1 with dual function of immunology and mineralization from the freshwater oyster (Hyriopsis cumingii Lea).

Shell and pearl formation in bivalves is a sophisticated biomineralization process that encompasses immunological and mineralization aspects, particularly during shell repair and the initial stages of pearl cultivation when a nucleus is inserted. Here, we describe a novel C-type lectin, HcLec1, isolated and characterized from the freshwater pearl mussel Hyriopsis cumingii Lea. Immune challenge, RNA interference (RNAi) experiments, ELISA, and antibacterial assays were employed to investigate the role of HcLec1 in innate immunity. We also established shell damage repair and pearl nucleus insertion models to examine the impact of HcLec1 on the biomineralization process in Hyriopsis cumingii Lea. In vitro calcium carbonate crystallization assays were conducted to explore the direct role of HcLec1 in calcium carbonate crystal formation. The HcLec1 gene sequence is a full-length cDNA of 1552 bp, encoding 240 amino acids. HcLec1 comprises an N-terminal signal peptide and a carbohydrate-recognition domain (CRD), with QPD (Gln-Pro-Asp) and MND (Met-Asn-Asp) motifs for polysaccharide binding. Tissue expression analysis showed that HcLec1 is predominantly expressed in the gill tissue of Hyriopsis cumingii Lea under normal conditions, and its expression is significantly elevated in both gill and pearl sac tissues following nucleus insertion for pearl cultivation (P < 0.05). After immune stimulation with Aeromonas hydrophila and lipopolysaccharides (LPS), HcLec1 expression levels significantly increased in both cases (P < 0.01), indicating a role in bivalve innate immunity. RNA interference (RNAi)-mediated knockdown of HcLec1 led to a significant decrease in the expression levels of immune-related genes (WAP, α2m, and Lyso) and mineralization-related genes (CA, CHS, Nacrein, and Pif) (P < 0.05). In animal models for shell damage and nucleus insertion in pearl cultivation, HcLec1 showed a consistent expression pattern, with an initial significant decrease followed by a marked increase, peaking at day 14 (P < 0.05). This suggests a role for HcLec1 in pearl formation and shell repair. The recombinant HcLec1 protein demonstrated binding affinity to LPS and PGN, a robust ability to agglutinate Escherichia coli, Staphylococcus aureus, Aeromonas veronii, and Aeromonas hydrophila, and significantly inhibited bacterial growth (P < 0.05). Moreover, rHcLec1 promoted calcite crystal formation in saturated calcium carbonate solutions and altered crystal morphology. The HcLec1 gene plays a pivotal role in both innate immunity and biomineralization in the triangle sail mussel. This study enhances our understanding of the functional diversity of C-type lectins and provides a foundation for future studies on shell repair and pearl growth.

Evaluation of Immunogenicity of Inactivated Whole-Virion Booster Vaccine against SARS-CoV-2 in a phase I, II Clinical Trial

Relevance. The development and production of own vaccine contributes to the growth of professional competencies of the scientific community, improvement and modernization of the country's industrial production.Aims. To evaluate the immunogenicity parameters of whole-virion vaccine against SARS-CoV-2 in a phase I, II clinical trial (up to day 180).Materials &amp; Methods. The clinical study included a phase I/II trial. In Phase I, a sequential allocation of subjects (1:1) by open-label method into two groups with a single administration of vaccine at dose 1 and at dose 2 was performed. In phase II (double-blind randomized), subjects were allocated to immunization with vaccine at doses 1, 2 or placebo in a 1:1:1 ratio. To evaluate the immunogenicity of an inactivated whole-virion booster vaccine against SARS-CoV-2 («BelCovidVac») in 135 healthy subjects (18–50 years of age in phase 1 and 18-60 years of age in phase 2) previously vaccinated/recovered from COVID-19. IgG concentration (BAU/mL) to the S-protein of SARS-CoV-2 was measured by the «SARS-CoV-2-IgG quantitative-ELISA-BEST» ELISA kit, viral neutralising activity of sera and proliferative activity of T-lymphocytes were evaluated, geometric mean (GMC) of IgG concentration was calculated, geometric mean of virus neutralizing activity titer (GMT compared to baseline. Statistical processing of the data was performed using the R programming language.Results. The values of fold change in IgG concentration to SARS-CoV-2 on day 42 and 90 in subjects who received «BelCovidVac» vaccine were significantly higher than those of subjects who received placebo (p=0,05, p=0,02, respectively). The value of the fold change in the titer of viral neutralizing activity in the study subjects who received «BelCovidVac» vaccine was significantly higher than the values of the subjects who received placebo (p=0,02). A statistically significant positive correlation (ρSp=0,51 p=0,0005) between the content of antigen-specific T-cells and the level of IgG after 28 days in the studied subjects and between the fold change in the titer of viral neutralizing activity of sera and the fold change in IgG concentration (ρSp = 0,40 p&lt;0,001) was established. On day 180, all groups of subjects show a decrease in antigen-specific IgG concentrations to the initial level.Discussion. At the time of the search for volunteers and the beginning of the clinical trial of the vaccine «BelKovidVac» the immune layer of the population had already been formed, which made the task of the researchers more difficult. But the vaccine «BelKovidVac» showed its immunogenicity, as evidenced by the results of the study.Conclusions. A significant increase in GMC IgG and GMT viral neutralizing activity of sera in the group vaccinated with «BelCovidVac» and no significant increase in GMC IgG and GMT viral neutralizing activity of sera in the placebo group were revealed, which is the evidence of immune response stimulation as a result of «BelCovidVac» vaccine administration and its efficacy.

A comparative study of the genetically improved Abbassa Nile tilapia strain (GIANT-G9) and a commercial strain in Egypt: growth vs. stress response

Selective breeding is a potent method for developing strains with enhanced traits. This study compared the growth performance and stress responses of the genetically improved Abbassa Nile tilapia strain (G9; GIANT-G9) with a local commercial strain over 12 weeks, followed by exposure to stressors including high ammonia (10 mg TAN/L), elevated temperature (37 °C), and both for three days. The GIANT-G9 showed superior growth, including greater weight gain, final weight, length gain, specific growth rate, and protein efficiency ratio, as well as a lower feed conversion ratio and condition factor compared to the commercial strain. The expression of growth hormone in the brain of the GIANT‐G9 increased significantly after 6 weeks, although it slightly decreased after 12 weeks. Growth hormone receptor 1 expression also increased significantly after 6 weeks. Muscle insulin-like growth factors (igf1 and igf2) levels up-regulated significantly only after 12 weeks in the GIANT‐G9. Under stress, serum enzymes (alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase (ALP)) were significantly higher in the GIANT‐G9, while the commercial strain had lower levels. No significant changes were observed in liver ALP activity among stressed strains. Under stress, the GIANT‐G9 exhibited marked upregulation of splenic Toll-like receptors (tlr2, tlr9, tlr21), myeloid differentiation primary response protein 88 (myd88), nuclear factor kappa B (nf-κB), interleukin (il) 1β, and il6. Notably, il6 expression was higher than il1β in the spleen, with the opposite pattern in the head kidney. In response to immune stimulation, globulin levels significantly increased in the GIANT‐G9 but with similar values to the stressed commercial strain. Myostatin expression increased in the spleen of the stressed GIANT‐G9. The commercial strain exhibited the best liver catalase and superoxide dismutase activities under stress, while the GIANT‐G9 showed increased liver glutathione-S-transferase (GST) activity after exposure to ammonia and temperature stress. Serum lysozyme activity increased in the stressed commercial strain and under temperature stress in the GIANT‐G9 but decreased under other stress conditions. Overall, the stressed commercial strain demonstrated higher survivability than the stressed GIANT‐G9. The study revealed significant interactions between strains and stress factors. The GIANT‐G9 exhibited higher growth rates but lower antioxidant and immune capacities compared to the commercial strain at the juvenile stage of life and production cycle.

A Novel Approach for In Vitro Testing and Hazard Evaluation of Nanoformulated RyR2-Targeting siRNA Drugs Using Human PBMCs.

Nucleic acid (NA)-based drugs are promising therapeutics agents. Beyond efficacy, addressing safety concerns-particularly those specific to this class of drugs-is crucial. Here, we propose an in vitro approach to screen for potential adverse off-target effects of NA-based drugs. Human peripheral blood mononuclear cells (PBMCs), purified from buffy coats of healthy donors, were used to investigate the ability of NA-drugs to trigger toxicity pathways and inappropriate immune stimulation. PBMCs were selected for their ability to represent potential human responses, given their likelihood of interacting with administered drugs. As proof of concept, a small interfering RNA (siRNA) targeting Ryanodine Receptor mRNA (RyR2) identified by the Italian National Center for Gene Therapy and Drugs based on RNA Technology as a potential therapeutic target for dominant catecholaminergic polymorphic ventricular tachycardia, was selected. This compound and its scramble were formulated within a calcium phosphate nanoparticle-based delivery system. Positive controls for four toxicity pathways were identified through literature review, each associated with a specific type of cellular stress: oxidative stress (tert-butyl hydroperoxide), mitochondrial stress (rotenone), endoplasmic reticulum stress (thapsigargin), and autophagy (rapamycin). These controls were used to define specific mRNA signatures triggered in PBMCs, which were subsequently used as indicators of off-target effects. To assess immune activation, the release of pro-inflammatory cytokines (interleukin-6, interleukin-8, tumor necrosis factor-α, and interferon-γ) was measured 24 h after exposure. The proposed approach provides a rapid and effective screening method for identifying potential unintended effects in a relevant human model, which also allows to address gender effects and variability in responses.

Instigator of Type 1 Diabetes Mellitus (T1D): Connecting Human Leukocyte Antigen (HLA) with Infant Gut Microbiome

Type 1 Diabetes Mellitus (T1D) is driven by the interplay between genetics and other factors, though the exact relationship between gut microbiome and T1D onset remains unclear. This study aims to investigate the connections between Human Leukocyte Antigen (HLA) genetics, gut microbiome, and their relationships to T1D seroconversion. Data from the Diabimmune website provided stool samples (n=1443) from children aged 16 to 36 months from Finland, Russia, and Estonia, categorized by HLA risk factors and seroconversion status. Statistical analyses using the Wilcoxon rank-sum test measure were conducted. High-risk HLA individuals generally had a lower relative abundance of beneficial bacteria such as Lachnospira. Predicted metabolic pathways PWY.5676 and PWY.7220/7222 had an increased abundance in low-risk HLA infants in multiple age groups. Prevotella was more abundant in seroconverted samples, while Ruminococcus, Hungatella, and Sutterella were more abundant in non-seroconverted samples. Shared pathways between different age groups such as GLYOXYLATE.BYPASS showed higher abundance in non-seroconverted samples, each with similar median abundance across different age groups. The findings suggest that gut microbiome dysbiosis, characterized by a lower abundance of beneficial SCFA-producing bacteria and the prevalence of pathways supporting bacterial persistence and immune stimulation, contributes to T1D onset. Future research should focus on advanced genomic and metabolomic analyses, functional assays, co-culture experiments, and developing predictive models using machine learning to assess autoimmune disease risk based on bacterial strains and HLA haplotypes

Targeted intra-tumoral hyperthermia using uniquely biocompatible gold nanorods induces strong immunogenic cell death in two immunogenically 'cold' tumor models.

Hyperthermia is an established adjunct in multimodal cancer treatments, with mechanisms including cell death, immune modulation, and vascular changes. Traditional hyperthermia applications are resource-intensive and often associated with patient morbidity, limiting their clinical accessibility. Gold nanorods (GNRs) offer a precise, minimally invasive alternative by leveraging near-infrared (NIR) light to deliver targeted hyperthermia therapy (THT). THT induces controlled tumor heating, promoting immunogenic cell death (ICD) and modulating the tumor microenvironment (TME) to enhance immune engagement. This study explores the synergistic potential of GNR-mediated THT with immunotherapies in immunogenically 'cold' tumors to achieve durable anti-tumor immunity. GNRs from Sona Nanotech Inc.™ were intratumorally injected and activated using NIR light to induce mild hyperthermia (42-48°C) for 5 minutes. Tumor responses were analyzed for cell death pathways and immune modulation. The immunogenic effects of THT were assessed alone and in combination with intratumoral interleukin-2 (i.t. IL-2) or systemic PD-1 immune checkpoint blockade. Immune cell infiltration, gene expression changes, and tumor growth kinetics were evaluated. THT reduced tumor burden through cell death mechanisms, including upregulated ICD marked by calreticulin exposure within 48 hours. By 48 hours, CD45+ immune cell levels were increased, including increased levels of immunosuppressive M2 macrophages. While THT led to innate immune cell stimulations highlighted by gene expression upregulation in the STING cGAS pathway and enhanced M1 and dendritic cell levels, tumor regrowth was observed within six days post-treatment. To enhance THT's immunogenic effects, the therapy was combined with intratumoral interleukin-2 (i.t. IL-2) or systemic PD-1 immune checkpoint blockade. Sequential administration of i.t. IL-2 post-THT induced robust CD8+ T-cell infiltration and led to sustained tumor regression in both treated and distant tumors, accompanied by the emergence of memory T cells. However, IL-2-induced immunosuppressive T-reg populations were also sustained to tumor endpoint suggesting that therapy could be further enhanced. Additionally, PD-1 expression, which was upregulated in CD8+ T cells by THT, was targeted with systemic PD-1 inhibition, further augmenting immune engagement within the TME. These combinatory treatments demonstrated synergistic effects, promoting durable anti-tumor responses and immune memory. Collectively, GNR-mediated THT effectively reduces tumor burden and remodels the TME, potentiating systemic immunity and enhancing the impact of complementary immunotherapies.

Tumor Vaccine Exploiting Membranes with Influenza Virus-Induced Immunogenic Cell Death to Decorate Polylactic Coglycolic Acid Nanoparticles.

Immunogenic cell death (ICD) of tumor cells, which is characterized by releasing immunostimulatory "find me" and "eat me" signals, expressing proinflammatory cytokines and providing personalized and broad-spectrum tumor antigens draws increasing attention in developing a tumor vaccine. In this study, we aimed to investigate whether the influenza virus (IAV) is efficient enough to induce ICD in tumor cells and an extra modification of IAV components such as hemeagglutinin (HA) will be helpful for the ICD-induced cells to elicit robust antitumor effects; in addition, to evaluate whether the membrane-engineering polylactic coglycolic acid nanoparticles (PLGA NPs) simulating ICD immune stimulation mechanisms hold the potential to be a promising vaccine candidate, a mouse melanoma cell line (B16-F10 cell) was infected with IAV rescued by the reverse genetic system, and the prepared cells and membrane-modified PLGA NPs were used separately to immunize the melanoma-bearing mice. IAV-infected tumor cells exhibit dying status, releasing high mobility group box-1 (HMGB1) and adenosine triphosphate (ATP), and exposing calreticulin (CRT), IAV hemeagglutinin (HA), and tumor antigens like tyrosinase-related protein 2 (TRP2). IAV-induced ICD cells enhance biomass-derived carbon (BMDCs) migration, antigen uptake, cross-presentation, and maturation in vitro. Furthermore, immunization with IAV-induced ICD cells effectively suppressed tumor growth in melanoma-bearing mice. The isolated cell membrane inherited the immunological characteristics from the ICD cells and elicited robust antitumor immune responses through decorating PLGA NPs loading with a tumor-specific helper T-cell peptide and supplemented with ATP in a hydrogel system. This study indicated a promising strategy for developing cell-based and personalized tumor vaccines through fully taking advantage of the immune stimulation mechanisms of ICD occurrence in tumor cells, IAV modification, and nanoscale delivery.

Spatiotemporal Dynamic Immunomodulation by Infection-Mimicking Gels Enhances Broad and Durable Protective Immunity Against Heterologous Viruses.

Despite their safety and widespread use, conventional protein antigen-based subunit vaccines face significant challenges such as low immunogenicity, insufficient long-term immunity, poor CD8+ T-cell activation, and poor adaptation to viral variants. To address these issues, an infection-mimicking gel (IM-Gel) is developed that is designed to emulate the spatiotemporal dynamics of immune stimulation in acute viral infections through in situ supramolecular self-assembly of nanoparticulate-TLR7/8a (NP-TLR7/8a) and an antigen with tannic acid (TA). Through collagen-binding properties of TA, the IM-Gel enables sustained delivery and enhanced retention of NP-TLR7/8a and protein antigen in the lymph node subcapsular sinus of mice for over 7 days, prolonging the exposure of vaccine components in both B cell and T cell zones, leading to robust humoral and cellular responses. The IM-Gel system with the influenza A antigen confers cross-protection against multiple influenza subtypes (H1N1, H5N2, H3N2, H7N3, and H9N2) with long-term immune responses. Combination of the IM-Gel with the SARS-CoV-2 spike protein also elicits strong cross-reactive antibody responses against multiple SARS-CoV-2 variants (Alpha, Beta, NY510+D614G, Gamma, Kappa, and Delta). The IM-Gel, as a programmable immunomodulatory material, provides a vaccine design principle for the development of next-generation universal vaccines that can elicit broad and durable protective immunity against emerging viruses.

An interferon-stimulated long non-coding RNA USP30-AS1 as an immune modulator in influenza A virus infection.

Long non-coding RNAs (lncRNAs) are essential components of innate immunity, maintaining the functionality of immune systems that control virus infection. However, how lncRNAs engage immune responses during influenza A virus (IAV) infection remains unclear. Here, we show that lncRNA USP30-AS1 is up-regulated by infection of multiple different IAV subtypes and is required for tuning inflammatory and antiviral response in IAV infection. Genetically inactivation of USP30-AS1 enhances viral protein synthesis and viral growth. USP30-AS1 is an interferon-stimulated gene, and the induction of USP30-AS1 can be achieved by JAK-STAT mediated signaling activation. The immune regulation of USP30-AS1 is independent of its proximal protein-coding gene USP30. In IAV infection, deletion of USP30-AS1 unleashes high systemic inflammatory responses involving a broad range of pro-inflammatory factors, suggesting USP30-AS1 as a critical modulator of immune responses in IAV infection. Furthermore, we established a database providing well-annotated host gene expression profiles IAV infection or immune stimulation.

Loss of DHX36/G4R1, a G4 resolvase, drives genome instability and regulates innate immune gene expression in cancer cells.

G-quadruplexes (G4s) are four-stranded alternative secondary structures formed by guanine-rich nucleic acids and are prevalent across the human genome. G4s are enzymatically resolved using specialized helicases. Previous in vitro studies showed that DEAH-box Helicase 36 (DHX36/G4R1/RHAU), has the highest specificity and affinity for G4 structures. Here, by mapping genome-wide DNA double-strand breaks (DSBs), we demonstrate that knockout (KO) of DHX36 helicase increases DSB enrichment at G4 sites and that the presence of the G4 motif is a significant mediator of genome instability at regulatory regions. The loss of DHX36 corresponds with the significant upregulation of NF-κB transcriptional programs, culminating in the production and secretion of proinflammatory cytokines. Loss of DHX36 expression results in an increase in the innate immune signaling stimulator of interferon response cGAMP interactor 1 ( STING1 ) expression and activation of genes involved in immune response pathways. Importantly, higher levels of DHX36 mRNA expression in human B-cell acute lymphoblastic leukemia correlate with improved overall survival relative to lower expression of DHX36 , highlighting its critical role in preserving genome integrity at a cellular level and in the context of cancer.

Peptides as Versatile Regulators in Cancer Immunotherapy: Recent Advances, Challenges, and Future Prospects.

The emergence of effective immunotherapies has revolutionized therapies for many types of cancer. However, current immunotherapy has limited efficacy in certain patient populations and displays therapeutic resistance after a period of treatment. To address these challenges, a growing number of immunotherapy drugs have been investigated in clinical and preclinical applications. The diverse functionality of peptides has made them attractive as a therapeutic modality, and the global market for peptide-based therapeutics is witnessing significant growth. Peptides can act as immunotherapeutic agents for the treatment of many malignant cancers. However, a systematic understanding of the interactions between different peptides and the host's immune system remains unclear. This review describes in detail the roles of peptides in regulating the function of the immune system for cancer immunotherapy. Initially, we systematically elaborate on the relevant mechanisms of cancer immunotherapy. Subsequently, we categorize peptide-based nanomaterials into the following three categories: peptide-based vaccines, anti-cancer peptides, and peptide-based delivery systems. We carefully analyzed the roles of these peptides in overcoming the current barriers in immunotherapy, including multiple strategies to enhance the immunogenicity of peptide vaccines, the synergistic effect of anti-cancer peptides in combination with other immune agents, and peptide assemblies functioning as immune stimulators or vehicles to deliver immune agents. Furthermore, we introduce the current status of peptide-based immunotherapy in clinical applications and discuss the weaknesses and future prospects of peptide-based materials for cancer immunotherapy. Overall, this review aims to enhance comprehension of the potential applications of peptide-based materials in cancer immunotherapy and lay the groundwork for future research and clinical applications.