Abstract: Background: Killed Whole Cell Genome-Reduced Bacteria (KWC/GRB), a versatile vaccine platform, can produce very low cost, thermostable, easily manufactured vaccines expressing complex immunogens that include potent immunomodulators. This system supports iterative optimization through a Design–Build–Test–Learn (DBTL) workflow aimed at enhancing immunogenicity. We applied this approach to developing an HIV-1 gp41 Membrane-Proximal External Region (MPER) vaccine using the scaffolded MPER antigen, 3AGJ, a recombinant heterologous protein engineered to mimic MPER structures recognized by broadly neutralizing monoclonal antibodies (bNAbs). Methods: Five KWC/GRB vaccines expressing versions of 3AGJ were designed, including versions linked to immunomodulators and multimers of the immunogen. Display on the surface of the bacteria was evaluated by flow cytometry using the broadly neutralizing monoclonal antibody 2F5. Outbred HET3 mice were vaccinated intramuscularly, MPER-specific an-tibody responses were assessed by ELISA, and the ability of the vaccines to induce neutralizing antibodies determined. Neutralization was measured against tier 1 and tier 2 HIV-1 pseudoviruses. Results: All five vaccines were strongly expressed on the bacterial surface and induced clear MPER-specific antibody responses in every mouse. About 33% of the animals showed detectable HIV-1 neutralization. Conclusion: A KWC/GRB 3AGJ scaffold-MPER vaccine can induce HIV-1 neutralizing antibodies. While improvements in the responses would be needed for a clinically useful vaccine, the findings provide an initial validation of the concept. There are many strategies that can be used to enhance and extend immune responses induced by KWC/GRB vaccines that can be employed to yield improved anti-HIV immune responses.

Abstract: Autoimmune rheumatic diseases arise when the immune system transitions from a flexible, self-regulating network into a metabolically and epigenetically fixed inflammatory attractor state. This review synthesizes emerging evidence that immune tolerance is governed by a coupled epigenetic–metabolic axis integrating mitochondrial fitness, chromatin accessibility, redox balance, and nutrient flux across lymphoid, myeloid, and stromal compartments. We examine how chronic cytokine signaling, hypoxia, and oxidative stress destabilize regulatory programs, imprint glycolytic effector states, and remodel enhancer landscapes, thereby sustaining autoreactive circuits even after inflammatory pathways are pharmacologically suppressed. Multi-omic and spatial analyses reveal that pathogenic chromatin architectures, persistent mitochondrial dysfunction, and intercellular metabolite exchange cooperate to establish self-sustaining inflammatory ecosystems in rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis, and Sjögren’s syndrome. We further highlight therapeutic strategies aimed at tolerance reprogramming, including metabolic correction, chromatin-targeted agents, CAR-Tregs, tolerogenic dendritic cells, and integrative biomarkers that quantify metabolic–epigenetic coherence. By reframing autoimmunity as a disorder of energetic and chromatin desynchronization rather than isolated immune activation, this review outlines a mechanistic path toward durable, drug-free remission through deliberate restoration of the molecular architecture that maintains immune self-recognition.

Abstract:

Within Sudden Infant Death Syndrome (SIDS) resides several primary phenomena; these include a state of immune immaturity, susceptibility to infection, and an inflammatory state. Most SIDS risk factors pertain in some way or another to higher risk of infection (prematurity, lack of breastfeeding, low or absent transplacental antibody, ethnicity, genetics, risky gene polymorphisms, poverty, etc.). Most SIDS cases display evidence of an inflammatory state (raised inflammatory markers and inflammatory cytokines). The pattern of inflammation is very similar to that observed following vaccination, which for achieving successful levels of protective immunity requires components that induce high reactogenicity. It is this reactogenicity which, under certain circumstances can cause immune paralysis. Immune paralysis leaves a vulnerable infant open to infection and systemic inflammatory response syndrome leading to shock. Such a mechanism is explored in this rapid review in the context of the aetiopathogenesis of SIDS.

Abstract:

Background: Oral food challenges (OFCs) are still the reference standard for confirming food allergy, yet the influence of previous anaphylaxis on challenge outcomes remains uncertain. Patients with a history of anaphylaxis are often considered at higher risk, which may affect clinical decision-making process. In our study, we sought to identify predictors of OFC failure stratified by previous history of anaphylaxis. Methods: We conducted a retrospective evaluation of standard-of-care OFCs to cow’s milk and hen’s egg white performed between January 2014 and July 2025 at the Pediatric Allergy Center. Eligible participants had suspected or confirmed IgE-mediated allergy to cow’s milk protein (CMP) or hen’s egg white protein (HEWP), and were classified according to the presence or absence of previous anaphylaxis to the challenged food. Clinical data, including age, gender, allergic comorbidities, family history of atopy, and levels of specific IgE (sIgE) were obtained from medical records and compared between groups. Open OFCs were conducted according to PRACTALL guidelines (2012/2024) under inpatient supervision with full emergency support. Specific IgE levels were measured using the ImmunoCAP® assay (Phadia, Uppsala, Sweden). Logistic regression models were used to assess the relationship between comorbidities, sIgE concentrations and OFC outcomes. Receiver operating characteristic (ROC) analysis evaluated diagnostic accuracy of sIgE levels in predicting OFC outcomes. Results: The analysis included 192 pediatric patients (median age 4.75 years, 66.1% male) undergoing OFCs: 106 to CMP and 86 to HEWP. Six challenges (3.1%) were inconclusive, giving 186 valid results. The overall OFC failure rate was 32.3% (30.2% to CMP and 34.9% to HEWP). Patients with a past history of anaphylaxis more frequently underwent cow’s milk challenges (p = 0.01). Atopic dermatitis was a more common comorbidity in those without prior anaphylaxis (p = 0.04), regardless of the trigger. In hen’s egg challenges, children with a history of anaphylaxis reacted to significantly lower cumulative doses (p = 0.03) than patients without. Logistic regression analysis identified atopic dermatitis as a predictor of OFC failure in children without prior anaphylaxis (p = 0.02), and asthma as a borderline predictor in those with previous systemic reactions (p = 0.05). Specific IgE levels correlated with OFC outcomes across allergens, with casein-sIgE showing the highest discriminative performance (AUC = 0.81) in children without previous anaphylaxis. Conclusions: Atopic dermatitis and asthma were identified as potential risk factors influencing OFC outcomes, depending on the patient’s history of anaphylaxis. The predictive accuracy of sIgE was different in groups stratified by presence of prior anaphylaxis. Casein-sIgE showed the highest diagnostic accuracy in children without previous severe reactions to CMP. A prior history of anaphylaxis was associated with increased allergen sensitivity but did not predict a higher risk of severe reactions during supervised OFCs. Presence of anaphylactic reactions in the past is an important consideration when selecting children for OFCs to CMP and HEWP, since it delineates distinct risk factors for challenge failure in these patient populations.

Abstract: NK cells are key components of the innate immune system, capable of recognizing and eliminating tumor or virus-infected cells and able to modulate both innate and adaptive immune responses. This makes NK cells attractive candidates for cancer immunotherapy, through passive approaches such as adoptive NK-cell transfer, or active approaches aimed at enhancing endogenous NK-cell activity in vivo. Promising results have emerged from preclinical studies and early-phase clinical trials. Nevertheless, the therapeutic efficacy of NK cell–based approaches is often limited by several factors, such as the poor NK cell persistence in vivo, the inefficient tumor infiltration, and the immunosuppressive milieu typical of the tumor microenvironment. The preclinical development of NK cell–based therapies relies largely on animal models. Humanized mouse models have evolved from early immunodeficient strains to more advanced systems incorporating human cytokines, which more effectively support NK cell development, maturation, and function. These models have substantially improved our understanding of human NK cell biology and enabled the evaluation of novel therapeutic strategies. However, further optimization is still required to better recapitulate the tissue-specific heterogeneity of human NK cells and their conditioning by the tumor microenvironment. In this review, we provide an overview of recent advances in the generation of humanized mouse models for NK cell–based cancer immunotherapy, discussing their advantages and limitations and highlighting how emerging technologies may contribute to the development of more predictive preclinical platforms.

Abstract: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) and next-generation al-logeneic T cell therapies are curative options for hematologic malignancies, but their effi-cacy is often limited by graft-versus-host disease (GvHD), a complex syndrome resulting from dysregulated donor–host immune interactions. GvHD is mediated by interconnected signaling pathways that regulate T cell activation, metabolic and epigenetic program-ming, tissue-specific migration, stromal-immune interactions, and fibrotic remodeling. Therapeutic agents targeting key pathways, including ruxolitinib (JAK1/2), ibrutinib (BTK/ITK), and belumosudil (ROCK2), can modulate inflammatory responses while pre-serving graft-versus-tumor (GvT) activity. Emerging technologies, such as CRISPR-based genome editing, single-cell and spatial multi-omics, and AI-driven network modeling, enable patient-specific mapping of signaling hierarchies, prediction of disease trajectories, and identification of actionable targets. Integration of these approaches supports precise modulation of immune circuits, offering alternatives to broad immunosuppression. These insights provide a framework for next-generation, individualized interventions that pro-mote durable immune tolerance without compromising anti-tumor immunity and high-light rational combination strategies to improve outcomes in allo-HSCT and allogeneic T cell therapies.

Abstract: Optic neuritis (ON) has been recognized since antiquity, but its modern clinical identity emerged only in the late 19th century and was definitively shaped by the Optic Neuritis Treatment Trial (ONTT). The ONTT established the natural history, visual prognosis, association with multiple sclerosis (MS), and therapeutic response to corticosteroids, building the foundation for contemporary ON management. Over the past two decades, ON has evolved from a seemingly uniform demyelinating syndrome into a group of biologically distinct disorders. The identification of aquaporin-4-IgG ON (AQP4-ON), myelin oligodendrocyte glycoprotein antibody–associated ON (MOG-ON), and double-negative ON has transformed diagnostic and therapeutic strategies. These subtypes differ in immunopathology, clinical course, MRI features, retinal injury patterns, CSF profiles, and long-term outcomes, making early and accurate differentiation essential. MRI provides key distinctions in lesion length, orbital tissue inflammation, bilateral involvement, and chiasmal or optic tract extension. Optical coherence tomography (OCT) offers complementary structural biomarkers, including severe early ganglion cell loss in AQP4-ON, relative preservation in MOG-ON, and variable patterns in double-negative ON. CSF analysis further refines diagnosis, with oligoclonal bands strongly supporting MS-ON. Together, these modalities enable precise early stratification and timely initiation of targeted immunotherapy, which is critical for preventing irreversible visual disability. Despite major advances, significant unmet needs persist. Access to high-resolution MRI, OCT, cell-based antibody assays, and evidence-based treatments remains limited in many regions, contributing to global disparities in outcomes. The pathogenesis of double-negative ON, reliable biomarkers of relapse and visual recovery, and standardized multimodal diagnostic thresholds remain unresolved. Future research must expand biomarker discovery, refine imaging criteria, and ensure equitable global access to cutting-edge diagnostic platforms and therapeutic innovations. Four decades after the ONTT, ON remains a dynamic field of investigation, with ongoing advances holding the potential to transform care for patients worldwide.

Abstract:

Atopic allergy is rising globally and placing a significant strain on healthcare systems, yet the understanding of the underpinning mechanisms of allergic sensitisation remains incomplete. Extracellular vesicles (EVs) have recently emerged as important mediators of immune modulation due to their diverse cargo and therefore may play a mechanistic role in allergic sensitisation development. Thus, this study investigated whether EVs released by activated dendritic cells (DCs) contribute to allergic sensitisation of the common egg allergen, ovalbumin (OVA). DCs were generated from human monocytes cultured with GM-CSF and IL-4, then stimulated with LPS and/or OVA. EVs were subsequently isolated using size-exclusion chromatography and added to freshly isolated naïve T cells at defined time points. T-cell responses were then analysed using spectral flow cytometry. The results highlight EVs derived from LPS or LPS+OVA-stimulated DCs enhanced IL-4 production and reduced IFN-γ production in naïve T cells from egg-allergic donors, indicating a shift toward a Th2 profile. In healthy donors, LPS-induced DC EVs also suppressed IFN-γ expression. Notably, EVs alone were insufficient to activate T cells without CD3/CD28 co-stimulation, suggesting that EVs may function as a “third signal” shaping T-cell polarisation. These findings highlight a potential role for DC-derived EVs in initiating allergic sensitisation.

Abstract: Background/Objectives: The current strategy for seasonal influenza prophylaxis relies on updating the vaccine components annually to account for the rapid antigenic drift of viruses and the low cross-protective efficacy of available vaccines. Mutant influenza viruses with truncated or deleted NS1 protein are known to stimulate cross-specific T-cell immune response and provide protection against heterosubtypic influenza A and B viruses. Methods: We generated NS1ΔC influenza A and B viruses with C-terminal NS1 deletions by reverse genetics. In a mouse model, we assessed the safety and immunogenicity of the B/Lee/NS1ΔC strain upon intranasal administration, as well as the mechanism of its cross-protective efficacy against sublethal B/Victoria and B/Yamagata challenge. We then investigated the potential of the intranasal Flu/UniVec vaccine–a trivalent formulation of NS1ΔC A/H1N1, A/H3N2, and B influenza viruses–to protect mice from lethal influenza infection with homologous, heterologous, and antigenically drifted influenza A and B viruses. Results: Intranasal immunization with the B/Lee/NS1ΔC strain was safe in mice. It activated cross-specific T-cell responses in the lungs and protected animals against heterologous challenge by reducing viral load, inflammation, and lung pathology. Immunization with the trivalent Flu/UniVec vaccine formulation improved survival and reduced weight loss and viral load upon challenge with A/H1N1pdm, A/H2N2, A/H5N1, and B/Victoria viruses. Conclusions: The trivalent intranasal replication-deficient Flu/UniVec influenza vaccine is a promising tool to improve seasonal influenza protection and preparedness for an influenza pandemic.

Abstract: CD80 and CD86 molecules are key costimulatory ligands expressed primarily on the surface of antigen-presenting cells (APCs). However, the presence of these molecules on T cells is currently uncommon and may be associated with various immunopathologies, such as allergic diseases. Literature data demonstrate that the presence of these molecules on T cells can polarize the immune response toward Th2, thereby enhancing inflammation. In our study, we found that all Type 2-dependent diseases we examined (atopic dermatitis, bronchial asthma) were associated with increased CD80 expression by immune cells. One of the modern and effective treatments for these diseases is genetically engineered biological therapy (GEBT). In this study, we found that in patients with severe allergy pathology, GEBT-associated cells exhibit changes in the percentage of cells expressing CD80 and CD86 on both monocytes and T cells.

Abstract: Background/Objectives: The Notch-ADAM17 pathway is a fundamental signaling mechanism where ADAM17, a disintegrin and metalloprotease, cleaves the Notch re-ceptor after the Notch receptor binds to a ligand. Crosstalk between Notch and AD-AM17 is often altered in pathological situations. Alterations in Notch2 expression, in particular, appear to be correlated with the onset of various autoimmune diseases. In primary Sjögren's disease (pSjD), an autoimmune disorder characterized by chronic inflammation, the role of ADAM17 has been extensively explored, but a correlation with Notch2 has not yet been evaluated. Methods: To analyze the gene and protein expression of Notch2 in pSjD and a possible correlation with ADAM17 expression and with the patient’s inflammatory grade, we employed an integrated co-detection pro-tocol to analyze salivary gland tissue sections by combining in situ hybridization (ISH) with immunohistochemistry (IHC). Results: combined ISH/IHC allows us to demon-strate an increased expression of Notch2 mRNA and protein in pSjD salivary glands (SGs) biopsies, that appears correlated with an increased expression of ADAM17, both in acinar and duct cells and in infiltrating lymphocytes. Notch2/ADAM17 expression is higher in biopsies of pSjD SGs characterized by a high degree of inflammation. Con-clusion: this work demonstrates the correlated expression in pSjD SGs of ADAM17, which plays multiple roles in the pathogenesis of SjD, and Notch2, widely considered a key player in various inflammatory mechanisms, offering a starting point for future therapeutic interventions to investigate.

Abstract: The literature describes cases of chronic poisoning by organophosphates: pesticides, flame re-tardants, and other industrial and household chemicals. The delayed effects of acute, subacute, subchronic and chronic poisoning manifest themselves years after exposure, with the effects of the toxic factor eventually masked by age-related diseases with an immunological component. The aim of this study was to investigate the immunological profile of blood cells in a cohort of elderly patients diagnosed with occupational pathology caused by subacute or (sub)chronic ex-posure to organophosphates in the 1980s. Analysis of well-being assessments revealed that patients in the control and organophos-phate-exposed groups had approximately equal levels of memory and thinking impairments, anxiety, melancholy and depression, and cognitive function assessments. However, the organo-phosphate-exposed group experienced significantly more balance and memory impairments, and more pronounced neurological symptoms, manifested by abnormal wrist reflexes, cranio-cerebral changes, impaired vibration and distal sensitivity, and depressed abdominal, ankle and plantar reflexes. The organophosphates-exposed group showed decreased red blood cell counts and hematocrit, white blood cell counts were unchanged in both groups. Lymphocyte phenotyping revealed a 1.5-2-fold increase in the relative and absolute levels of circulating plasma cell precursors with the CD19+IgD–CD27++ phenotype, an absolute count of cells with the CD19+CD27++CD38++ phenotype, and an absolute count of TEMRA CD4+ T cells in the organophosphate-exposed group of patients. However, the proportion of CD56+CD57+ cells of the EM4 (CD27–CD28+) CD8+ T cell subset was more than 2-fold reduced. The identified changes indicate a disruption of immune regulation in elderly people with occu-pational pathology, which along with indicators of the neurological and cognitive status of pa-tients can shed light on the pathogenesis of late pathology and serve as important markers in differential diagnosis.

Abstract: Prison staff are a high-risk population for latent tuberculosis infection (LTBI) and psy-chological distress due to their occupational environment. Considering the immuno-modulatory role of interleukin-22 (IL-22) and the limited evidence in this context, this cross-sectional study investigated the association between LTBI, serum IL-22 levels, and psychological distress among prison staff in São Paulo State, Brazil. Seventy-nine participants (24 females, 55 males) were evaluated using the QuantiFERON®-TB Gold Plus assay, the Depression, Anxiety, and Stress Scale-21 (DASS-21), and enzyme-linked immunosorbent assay (ELISA) for IL-22. LTBI prevalence was 34.2%, with symptoms of depression, anxiety, and stress present in 13.92%, 22.78%, and 20.25% of partici-pants, respectively. No significant differences in psychological scores were observed between LTBI-positive and negative groups. IL-22 levels tended to be higher in LTBI-positive individuals and were positively associated with interferon-gamma (IFN-γ) responses to TB-specific antigens, while inversely related to mito-gen-stimulated responses. These findings suggest a selective immunomodulatory role for IL-22, enhancing pathogen-specific responses while potentially downregulating generalized immune activation. Depression and stress scores were also positively asso-ciated with IFN-γ levels. These findings highlight IL-22 as a potential biomarker of immune-psychological dynamics in LTBI and support integrated mental and immune health monitoring in high-risk occupational environments.

Abstract:

Streptococcus pneumoniae contributes significantly to morbidity, mortality, and healthcare costs worldwide due to severe Invasive Pneumococcal Disease (IPD), particularly among young children and vulnerable populations. This review critically examines the current state of pneumococcal disease epidemiology, the evolution of vaccine strategies, and persistent challenges to achieve global control of the disease. The implementation of Pneumococcal Conjugate Vaccines (PCVs) has yielded substantial public health gains, establishing herd immunity and sharply reducing vaccine-type IPD incidence. However, this success has been fundamentally challenged by serotype replacement, where non-vaccine serotypes have subsequently emerged to cause a significant proportion of the residual disease burden. This epidemiological shift has necessitated the development and deployment of higher-valency PCVs (PCV15, PCV20, and PCV21) to expand serotype coverage. Furthermore, optimal protection requires personalized strategies for high-risk cohorts where vaccine effectiveness can be compromised. In this context, the review details how pneumococcal vaccination - and particularly PPSV23 - serves as an indispensable diagnostic tool to evaluate a broad spectrum of Inborn Errors of Immunity (IEI) and in particular humoral defects. Diagnostic challenges are strained by non-standardized assays and the limited panel of unique serotypes available for testing in the PCV era. The scientific priority is now the development of universal protein-based vaccines, to provide protection against all serotypes and non-encapsulated strains by targeting conserved virulence factors. This integrated approach, combining expanded PCV coverage with novel vaccine technology, is essential to mitigate the ongoing public health burden of pneumococcal disease.

Abstract: Background/Objectives: Allergen immunotherapy (AIT) with chemically modified allergoids represents a promising approach for the treatment of allergic rhinoconjunctivitis. This post-hoc analysis investigated the clinical efficacy of a mannan-conjugated birch pollen allergoid (T502) by comparing outcomes in patients who received placebo in one pollen season and active treatment in the following year. Methods: Data were derived from four randomized clinical trials (EudraCT Numbers. 2018-002522-23, 2020-004126-32, 2021-002252-36, 2022-004082-20) conducted over four consecutive years. Two independent patient groups were analyzed, each receiving placebo in the first year and subcutaneous T502 (10,000 mTU/mL) in the subsequent year. Daily symptom score (dSS), daily medication score (dMS), and combined symptom and medication score (CSMS) were assessed during the birch pollen season each April. Results: In the first comparison between 2020 and the early start in January 2021, treatment with T502 led to a 43.79% reduction in dSS (p = 0.000), a 71.43% reduction in dMS (p = 0.001), and a 42.1% reduction in CSMS (p = 0.001), all statistically significant. In the second comparison between 2022 and a late start in March 2023, the reduction in dMS was 74.47% (p = 0.001), whereas reductions in dSS (1.47%, p = 0.898) and CSMS (20.3%, p = 0.057) were not statistically significant. Conclusions: Subcutaneous immunotherapy (SCIT) with T502 significantly reduces allergic symptoms and medication use, particularly when all injections are completed before the onset of the birch pollen season in April. These findings highlight both the clinical value of T502 and the importance of optimized treatment timing.

Abstract:

Background/Objectives: For decades, only native allergen extracts with a high incidence of adverse drug reactions (ADRs) were available for allergen-specific immunotherapy (AIT) - in general administered subcutaneously - to treat allergy to feline epithelia. Modified allergen extracts are a promising alternative to reduce the number of ADRs. The purpose of this study was to collect data on the safety of the depigmented allergoid from cat under real-world conditions in clinical routine. Methods: The study was designed as voluntary non-interventional post-authorization safety study (NIS-PASS) with specific focus on the occurrence of adverse events (AEs) – respectively ADRs – upon injections and on the assessment of the influence on quality of life (QoL). Period of observation was the initial phase (up-dosing) of the AIT. Results: 101 patients were included, of whom 91 patients were treated with the depigmented allergoid from cat. Regardless of the age group, around 50% of patients reported ADRs, which were mainly delayed local reactions (LR). Other ADRs occurred only sporadically without persistent impairment for the patients. The incidence of ADRs neither differed significantly between quick and conventional up-dosing regimens nor between adult and adolescent patient groups. The QoL data revealed no significant changes in any domains of the SF-12 questionnaire for the observation period of 12 weeks. Conclusions: Overall, subcutaneous allergen immunotherapy (SCIT) with depigmented allergoid from cat is a well-tolerated and safe treatment option for patients with cat allergy.

Abstract: Autoimmune rheumatic diseases (AIRD) are heterogeneous, relapsing–remitting disorders in which early diagnosis, flare prediction, and individualized treatment selection remain critical unmet needs. Recent advances in multimodal biomarkers—including serological and inflammatory markers, quantitative imaging (ultrasound/MRI), -omics signatures (e.g., interferon- and B-cell–related programs), and digital phenotypes from wearables and smartphones—can now be fused through AI pipelines to enhance phenotyping, risk stratification, and treatment-response modeling. This review synthesizes recent advances across three interconnected domains: (i) imaging artificial intelligence (AI), which standardizes the quantification of synovitis, erosions, and microvascular changes; (ii) omics-based stratification approaches in systemic lupus erythematosus (SLE) and related AIRD; and (iii) remote, patient-generated data streams that extend and complement traditional clinic-based assessments. We emphasize implementation science, highlighting registry-enabled infrastructures (e.g., ACR RISE), federated learning to preserve privacy across sites, and modern validation standards (TRIPOD+AI, PROBAST+AI, CONSORT-AI/SPIRIT-AI). Finally, we address equity and drift-monitoring, underscoring the need for continuous recalibration across ancestry, sex, age, and care settings. Collectively, these innovations are transitioning precision rheumatology from conceptual promise toward pragmatic, clinic-embedded deployment.

Abstract:

The C1858T PTPN22 (R620W) variant has been implicated in the pathogenesis of several autoimmune disorders and represents a promising immunotherapeutic target for Type 1 diabetes. We have been implementing a novel immunotherapeutic approach based on the use of lipoplexes that deliver siRNA duplexes. Efficacy and safety of lipoplexes halting variant expression was demonstrated in the peripheral blood of patients in vitro. According to regulatory authorities in Europe preclinical safety and efficacy must be ascertained in vivo in appropriate animal models before undertaking clinical investigations. In the light of the foregoing the aim of this study was to verify that lipoplexes against the murine Ptpn22-R619W, equivalent to the human PTPN22-R620W, could be used for animal experimentation. The murine fibroblast cell line L929 was transfected with the PF62-pLentiPtpn22-R619W plasmid. We designed siRNA duplexes specific for the Ptpn22-R619W allele and formulated them into cationic lipoplexes in order to halt the variant expression in the transfected L929 cell line. Transfection of fibroblasts expressing R619W using lipoplexes resulted in efficient silencing at 100 pmol siRNA after 48 hours post-transfection reaching higher significant knock‑down after 72 hours. Lipoplexes efficiently suppress the pathogenic Ptpn22 variant expression in vitro, supporting the feasibility of a pre‑clinical platform for in vivo lipoplexes testing in CRISPR‑engineered NOD/ShiLtJ mice carrying the R619W mutation.

Abstract: While the methodological principle “correlation does not imply causation” serves as a crucial safeguard in scientific inquiry, its overly restrictive application can impede legitimate investigative pathways and hinder the proper evaluation of emerging evidence. This phenomenon has been clearly observable in the COVID-19 vaccine safety discussions, where serious adverse events with temporal proximity to vaccination have been frequently rejected without comprehensive examination. This work invites reflection on how the inappropriate application of this methodological principle can obstruct scientific inquiry into adverse event surveillance, potentially affecting scientific credibility. This work applies Kuhnian paradigm theory to analyze how institutionalized frameworks operate simultaneously as mechanisms for knowledge advancement and as barriers to conceptual innovation, particularly in politically contested domains such as vaccine safety evaluation. Additionally, a case report is analyzed using Hill´s criteria for causality, in which an immunohistochemical method was used for the first time to evaluate whether mRNA-based vaccines against SARS-CoV-2 contributed to the cause of death. Results showed that three of the nine criteria demonstrated strong evidence of causality, four were partially fulfilled, and two were not fulfilled. Overall, the report provides approximately 55% causal evidence. This suggests, but does not prove, a causal relationship between the BNT162b2 vaccine and multifocal necrotizing encephalitis. The absence of the nucleocapsid protein does not provide definitive evidence to demonstrate a vaccine origin. An analytical method is described that can distinguish between the wild-type SARS-CoV-2 spike protein and the recombinant spike protein expressed following mRNA vaccination.

Abstract: Background Interferon Regulatory Factor 5 plays an important role in the regulation of innate immune responses by amplifying the Nuclear Factor κB response, which is critical in gout inflammation. Furthermore, the rs4728141 polymorphism C allele was associated with both increased IRF5 expression and susceptibility to gout. We examine the association between rs4728141 and cytokine production in response to various Toll-Like Receptor ligands and describe the transcriptomic and proteomic changes observed in patients with gout and controls in relation to this polymorphism. Materials and Methods We examine the transcriptome of freshly isolated peripheral blood mononuclear cells (PBMCs) from 93 healthy donors and 63 gout patients as well as serum inflammatory proteome in 197 control and 195 gout samples. 24-hour stimulation experiments of freshly isolated human PBMCs were performed, followed by RNA-sequencing in gout patients and cytokine production measurement by ELISA in healthy donors and gout patients. Results The rs4728141 C allele was associated with increased IL-1β expression in unstimulated PBMCs of controls, but not in gout. No association between the polymorphism and serum inflammatory proteome was found. As expected, an increased IRF5 expression was observed in stimulated PBMCs of rs4728141 C allele carriers in response to several stimulations. Interestingly, IL-1β production was specifically enhanced in association to the rs4728141 C allele when cells were stimulated with palmitate with or without monosodium urate crystals. Conclusions The recently identified gout risk allele C rs4728141, which maps to the vicinity to the IRF5 gene is associated with elevated proinflammatory responses and this was specifically observed in response to gout-relevant stimulations such as palmitate in presence or absence of monosodium urate crystals. This pattern of cytokine production shows a functional impact of rs4728141 in gout through altered IL-1β production, a main driver of inflammation in gout.