Medicine and Pharmacology

Abstract:

Background/Objectives: Therapeutic cancer vaccines can generate measurable immunity without tumor regression. Adjuvants initiate innate activation, but antitumor efficacy also requires antigen presentation, lymphoid priming, effector differentiation, tumor trafficking, cancer-cell recognition and killing, and persistence. We compared the magnitude, duration, and downstream breadth of clinically used adjuvant responses with live vaccination. Methods: We reanalyzed transcriptomic data from six randomized human vaccine studies and one controlled mouse experiment; one human study sequenced six sorted leukocyte populations. Twenty-three prespecified gene sets spanning innate ignition, antigen presentation, adaptive differentiation, trafficking, and effector programs were scored by participant-paired, baseline-adjusted comparisons with matched controls. One trial sampled placebo, adjuvanted, unadjuvanted, and live attenuated vaccines daily. Results: AS01B, AS01E, AS03, and MF59 increased type I interferon by 9.6, 8.6, 7.5, and 3.2 percentile points at 24 hours; AS04 and aluminum salt remained at control levels. Yellow fever 17D sustained interferon through day 7; MF59 resolved by day 3 and AS01/AS03 by day 7. Antigen-presenting-cell activation and MHC class I machinery rose with interferon. Apart from a small day-7 germinal-center/plasmablast signal in two AS03 trials, no reproducible sustained downstream program was detected in blood; acute cytotoxic and natural-killer-cell decreases mainly reflected blood-cell composition. Conclusions: Adjuvants produce robust ignition, but these blood datasets do not establish the later cellular functions required for tumor-cell killing. Duration is one candidate determinant of this transition. Cancer-vaccine trials should measure ignition together with compartment-appropriate priming, trafficking, cytotoxic function, and persistence across repeated doses.

Abstract: Background: Sequential cytokine conditioning of human peripheral blood mononuclear cell (PBMC)-derived cultures can generate heterogeneous extracellular outputs whose biological identity is easily oversimplified. Methods: We retrospectively integrated archived records from an IL-4/GM-CSF-supported differentiation sequence, subsequent IL-10/TGF-β1 conditioning, and recovery of a multicomponent extracellular preparation. Each analytical domain retained its documented experimental unit and was linked within a source-to-output framework. Results: One represented Annexin V/7-AAD output comprised 26,071 events with 99.66% viability. Archived cell-associated regions were integrated descriptively. The recovered preparation showed a heterogeneous particle-associated signal predominantly at approximately 170–240 nm together with reported detection of CD9, CD63, CD81, and TSG101. An aggregate summary of ten conditioned cultures reported mean extracellular values of 25,320.16 pg/mL IL-10, 659.35 pg/mL IL-4, 99.61 pg/mL IL-6, and 3.01 pg/mL TNF-α; IL-10 and IL-4 were conditioning inputs and were therefore interpreted as final-environment attributes. Conclusions: The study establishes a traceable source-to-output framework for a human PBMC-derived multicomponent extracellular preparation, preserves a biogenesis-neutral identity for its vesicular and non-vesicular components, and provides a transferable evidence-to-claim pathway for tolerance-oriented biologic development.

Abstract: Ragweed (Ambrosia artemisiifolia) pollen allergy is a major and growing public health problem worldwide, with clinical manifestations ranging from rhinitis and conjunctivitis to asthmatic episodes and anaphylaxis. The major ragweed allergen, Amb a 1, exhibits high IgE-binding potential and triggers type 1 hypersensitivity reactions. Although allergen-specific immunotherapy (AIT) effectively induces long-term immune tolerance, conventional extract-based formulations face critical limitations including batch-to-batch variability, heterogeneous allergen composition, IgE-mediated adverse reactions, inconsistent immunogenicity, and poor patient compliance. This critical re-view synthesizes and evaluates published evidence on current synthetic vaccine strategies for ragweed AIT, with emphasis on their design, immunological performance, and quality control considerations. This review critically evaluates current approaches to synthetic vaccine development for ragweed AIT, with particular focus on pep-tide-based constructs, recombinant proteins, DNA vaccines, and adjuvant-enhanced formulations, alongside corresponding quality control strategies. The available evidence indicates that synthetic vaccines provide a more rational, molecularly characterized trajectory for producing optimized AIT products with improved safety profiles, enhanced clinical efficacy, reduced injection frequency, and improved batch standardization relative to extract-based immunotherapy. These advances highlight the potential of synthetic vaccine platforms to support personalized immunotherapy and more effective disease-modifying treatment of allergic disorders.

Abstract: The G protein-coupled seven-transmembrane receptor, CXC chemokine receptor 5 (CXCR5), is predominantly expressed on B cells located in the secondary lymphoid tissues, follicular helper T cells, and lymphoma cells. Binding to its ligand, CXCL13, mediates cell migration and regulates lymphocyte trafficking. Aberrant CXCL13/CXCR5 expression and signaling have been implicated in tumor progression, autoimmune diseases, and chronic inflammatory disorders. Therefore, specific mAbs against CXCR5 are expected to be useful for diagnosis and therapeutic applications. In this study, novel anti-human CXCR5 mAbs (Cx₅Mabs) were developed through flow cytometry-based high-throughput screening. One clone, Cx₅Mab-6 (IgG_2b, κ), recognized CXCR5-overexpressed Chinese hamster ovary (CHO)-K1 cells but did not react with the other 5 CXCR receptors-overexpressed CHO-K1 cells in flow cytometry. Additionally, Cx5Mab-6 recognized endogenous CXCR5 in the human Burkitt lymphoma Raji cell line. The dissociation constant (K_D) values of Cx₅Mab-6 for CHO/CXCR5 and Raji were 3.4 × 10⁻⁹ M and 1.2 × 10⁻10 M, respectively. Furthermore, Cx₅Mab-6 is useful for Western blotting and can detect CXCR5 in human lymphoma tissue by immunohistochemistry. These findings suggest that Cx₅Mab-6 is versatile for basic research and has potential applications in clinical diagnosis.

Abstract: Entry of SARS-CoV-2 via mucosal surfaces of the upper respiratory tract including the oral cavity may be influenced by innate and adaptive immunity. We aim to determine whether antibodies in secretions might influence the SARS-CoV-2 burden and thus the severity of infection. Blood and stimulated oral fluid (SOF) samples were collected at recruitment (d0) from 173 patients, and sequentially at d14, d30 and d90 from 52 SARS-CoV-2 RT-PCR-confirmed patients. Anti-SARS-CoV-2 spike antibodies of IgG, IgA and SIgA isotypes were detected by ELISA. SARS-CoV-2 RNA copies were quantified by RT-PCR. SARS-CoV-2 RNA copies become negative by d14 in most subjects. At d14, 5/18 SOF samples continued to be RT-PCR positive. Serum/SOF IgG antibodies were detected in all patients; IgG/IgA/SIgA antibodies increased by d14 and decreased by d90. At d0 and d14, SOF RNA copies were negatively correlated with SOF/serum IgG and with SOF IgA/SIgA antibodies. Higher SOF antibodies were significantly associated with a more rapid decline in SARS-CoV-2 burden. SARS-CoV-2 RNA copies and spike-specific antibodies show differential trends during the major and minor COVID-19 waves in India. Avidity indices for SOF IgG/IgA antibodies declined by d90. Taken together our data suggests a potential functional role for SOF anti-SARS-CoV-2 spike antibodies in reducing the SARS-CoV-2 burden.

Abstract: Background/Objectives: Placebo responses can substantially influence patient-reported outcomes in clinical trials and are particularly relevant in allergen immunotherapy (AIT), where treatment efficacy is commonly assessed using symptom- and medication-based endpoints. We conducted an exploratory post hoc cross-study comparison of birch pollen-allergic participants who received pre-seasonal placebo injections in two randomised, double-blind, placebo-controlled AIT trials with participants from a third trial in whom the planned study injections were not administered before the corresponding birch pollen season. Methods: Data were pooled from three prospective clinical studies evaluating the mannan-conjugated birch pollen allergoid EP-088-T502: T502-SIT-020, T502-SIT-045, and T502-SIT-068. Placebo-injection cohorts from studies T502-SIT-020 and T502-SIT-045 were compared with the injection-free cohort from study T502-SIT-068. Clinical outcomes included the Combined Symptom and Medication Score (CSMS), daily Symptom Score (dSS), and daily Medication Score (dMS). Results: The three comparison cohorts comprised 321 participants: 161 in the injection-free cohort and 160 in the placebo-injection cohorts. Participants in the injection-free cohort had significantly higher symptom and medication scores during the secondary fixed 1–30 April analysis period than participants in the placebo-injection cohorts. Median CSMS was 1.97 in the injection-free cohort compared with 1.32 and 0.86 in the placebo-injection cohorts. Likewise, median dSS (1.08 vs. 0.79 and 0.59) and dMS (0.87 vs. 0.33 and 0.25) were higher in the injection-free cohort. Pairwise comparisons indicated statistically significant between-cohort differences. Conclusions: In this non-randomised, non-concurrent cross-study comparison, symptom and rescue-medication scores were lower in the placebo-injection cohorts than in the injection-free cohort. Because the cohorts were derived from separate studies conducted in different years, these findings are hypothesis-generating and do not establish a causal effect of placebo injections. Trial-related contextual factors and other cross-study differences should be considered when interpreting the results.

Abstract: Type 1 Diabetes Mellitus (T1DM) is preceded by a prolonged asymptomatic autoimmune phase during which islet autoantibodies represent the earliest detectable biomarkers of disease. Although radiobinding assays (RBA) remain the reference method for autoantibody detection, their use is limited by the requirement of radioactive reagents, specialized facilities and high operational costs, highlighting the need for sensitive, non-radiometric assays suitable for large-scale screening. In this study, we developed a bridge Fluorophore-Linked Immunosorbent Assay (b-FLIA) for the simultaneous detection of glutamic acid decarboxylase autoantibodies (GADA) and zinc transporter 8 autoantibodies (ZnT8A) using a recombinant ZnT8/GAD65 chimeric antigen expressed in a baculovirus–insect cell system. Assay performance was evaluated using sera from healthy controls and diabetic patients previously characterized by RBA. The b-FLIA achieved an area under the receiver operating characteristic curve of 0.9867, with 90.5% sensitivity and 100% specificity for the detection of ZnT8A and/or GADA. Furthermore, the assay showed an almost perfect agreement with RBA (κ = 0.912). These findings demonstrate that the combination of a baculovirus-expressed ZnT8/GAD65 chimeric antigen with a bridge fluorescence immunoassay constitutes a practical and sensitive platform for multiplex detection of diabetes-associated autoantibodies, with potential application in routine laboratories and future population-based screening programs.

Abstract: Background: Pharmacist involvement in immunization has expanded as a strategy to improve access to vaccines. In Costa Rica, private community pharmacies operate under a regulatory framework for vaccination services, yet little evidence exists on the private dispensing of administrable vaccines. The aim of this study was to describe the dispensing activity of administrable vaccines in private community pharmacies in Costa Rica by units, economic value, vaccine category and provincial distribution during MAT February 2025–MAT February 2026. Methods: A descriptive, retrospective, national study was conducted using an aggregated secondary IQVIA database of the Costa Rican private pharmaceutical market. Dispensed units, economic value in USD, percentage growth and share by vaccine category and province were analyzed; no individual patient, professional, establishment or nominal records were used. Results: Dispensed units rose from 162,664 to 217,387 (+33.6%). Economic value increased from USD 8,700,809 to USD 13,416,717 (+54.2%). Influenza was the highest-volume category in MAT February 2026 (87,064 units). Yellow fever showed the greatest growth, rising from 22,281 to 70,133 units and from USD 2.7 million to USD 7.0 million. HPV ranked second in economic value (USD 3.6 million). San José concentrated 63.0% of units and 52.9% of national economic value. Conclusions: The private market for administrable vaccines in Costa Rican community pharmacies expanded substantially in volume and economic value, driven mainly by yellow fever, while influenza led in units and HPV retained a major economic weight. These findings position private pharmacies as an active vaccine dispensing channel but do not allow estimation of vaccination coverage or doses actually administered.

Abstract: Encapsulating protein subunit vaccines in biodegradable microparticles (MP) can increase induction of long-term protective mucosal and systemic antibodies after respiratory administration but requires incorporation of a mucosal immunostimulant. We previously found that incorporating complement peptide-derived immunostimulant-02 (CPDI-02) with MP-encapsulated LPS-free OVA through surface modification of the same ~1 µm PLGA 50:50 MP greatly increases mucosal and systemic OVA-specific antibodies in young, naïve female C57BL/6 mice at 14 and 90 days post-intranasal administration. Here, we directly compared effects of incorporating CPDI-02 or inactive scCPDI-02 with MP-encapsulated LPS-free OVA by separate encapsulation (SE), co-encapsulation (CE), or surface modification (SM) on induction of OVA-specific IgA, IgM, and IgG antibody secreting cells (ASCs) in the lungs and spleen, IgA antibodies in nasal lavage fluid, IgA and IgG antibodies in bronchoalveolar lavage fluid, and IgG subclasses in the serum at 14 days post-intranasal administration. We found that SE incorporation induced greater or similar levels of mucosal and systemic OVA-specific ASCs and mucosal IgA and IgG titers but greater titers of systemic IgG subclasses than CE or SM incorporation versus inactive scCPDI-02. Thus, given that surface modification with CPDI-02 induces high titers of OVA-specific systemic and mucosal antibodies at 14 and 90 days post-intranasal immunization under the same experimental conditions, separate encapsulation of CPDI-02 is expected to more broadly induce long-term mucosal and systemic antibodies against MP-encapsulated protein vaccines than co-encapsulation and surface modification after intranasal and possibly other routes of mucosal immunization.

Abstract: Immunopharmacology spans immunostimulation, immunosuppression, and immunomodulation across oncology, transplantation, autoimmunity, allergy, and infection, yet it lacks a unifying organising framework comparable to the hallmarks of cancer in oncology; the immunological hallmark schemes that exist are disease- or therapy-specific rather than general. We present the Immunity Hallmarks Matrix, a two-axis conceptual framework constructed by structured expert synthesis. The first axis is a biological spine of fifteen hallmarks of the immune response, each a discrete and mechanistically defined capability, grouped into three functional clusters — Recognition and Activation, Effector and Memory, and Regulation, Tolerance and Context. The second axis classifies every therapeutic strategy by its intent toward a hallmark: to Enhance, Diminish, or Alter that capability. The organising principle is therapeutic intent rather than molecular identity: the same biological hallmark may be acted upon in opposite directions in different clinical settings, illustrated by the dual-coding of checkpoint control and by the enhance-early, diminish-late course of a viral illness. The crossing of the two axes yields a grid whose intersections are either coherent therapeutic targets, structurally empty (not a coherent goal), or as-yet-unaddressed white space that the framework exposes as hypothesised opportunity. The matrix is offered as a navigable, generative conceptual grammar for immunopharmacology with educational and research applications, and it subsumes existing descriptive frameworks as special cases. It is a conceptual synthesis, not a clinical instrument or a prescribing tool.

Abstract: Hepatitis B virus (HBV) remains highly endemic in sub-Saharan Africa, where perinatal and early childhood transmission contribute substantially to chronic infection. Although routine infant vaccination is widely implemented, birth-dose coverage remains inconsistent in many settings. This study carried out in strict compliance with ethical rules, evaluated serological markers of HBV exposure and vaccine-induced immunity among fully vaccinated infants in Cameroon. It was a cross sectional study involving 9 to 15 months old hepatitis B fully vaccinated and HIV negative infants living in the towns of Douala and Yaoundé. Infants born to mothers known to be HBsAg-positive were excluded. Serum samples were tested for HBV serological markers, using lateral flow immunochromatography [1] techniques and indirect elisa [2,3]. Whereas 84.85% (56/66) of theses infants achieved seroprotective anti-HBs levels, 16.67% (11/66) was Anti-HBc seropositive. Overall, 63.64% of these anti-HBc seropositive infants were significantly less likely to demonstrate seroprotection compared with anti-HBc seronegative infants (adjusted OR 30.3; 95% CI 5.58–164.75). These results that highlights the association between anti-HBc positivity and reduced seroprotection, warrants further investigation, including molecular testing to distinguish passive maternal antibody transfer from occult or resolved infection. Strengthening perinatal HBV prevention strategies remains critical to achieving elimination targets.

Abstract: Autoimmune and autoinflammatory diseases represent two major, closely related categories of immune-mediated disorders. Patients with mixed autoinflammatory–autoimmune phenotypes often experience substantial diagnostic delay and suboptimal treatment owing to overlapping clinical features and heterogeneous pathogenic mechanisms. In this article, we review current approaches to the diagnosis and management of mixed autoinflammatory and autoimmune disorders, highlight illustrative clinical scenarios and available therapeutic strategies, and discuss emerging artificial intelligence-based methodologies for precision diagnosis, risk stratification, and prognosis in this complex disease spectrum.

Abstract: Allergen-specific IgE multiplex serology enables the molecular characterization of sensitizations in large populations. This study analyzed results from 21,121 canine sera collected in the USA during 2024–2025 using the Pet Allergy Xplorer (PAX) 238-allergen platform to determine sensitization prevalence and profiles. Sensitization to at least one allergen and to at least one environmental allergen was detected in 90.7% and 86.2% of dogs, respectively. House dust mites (HDM) were the predominant allergens (79.5%), followed by Hymenoptera venoms (39.8%), meats (21.9%), weeds (21.5%), and tree pollens (18.6%). Sensitization to the Dermatophagoides farinae HDM was the most prevalent (76.7%). Unsupervised clustering of 16,366 sensitized dogs identified six biologically coherent profiles centered on flea saliva, mammalian serum albumins/IgGs, honeybee venom, HDM group-2 allergens, storage mites, and PR-10-family allergens. A second, pollen/plant-food-focused analysis found clusters associated with Parietaria Par j 2, ragweed, granule-bound starch synthases, and PR-10 allergens, as well as a minor Phl p 6-associated grass pollen profile. Altogether, these results define the molecular sensitization repertoire of allergic dogs in the US and provide the background for interpreting IgE serological results and optimizing allergen selection for immunotherapy.

Abstract: Lipedema affects an estimated 11 to 12% of women and is characterized by bilateral, symmetric, painful enlargement of the limb subcutaneous fat that resists dietary weight loss. Its primary substrate is increasingly understood to be adipo-vascular and connective: genome-wide association studies implicate adipose, vascular, and extracellular-matrix loci rather than immune genes, and the affected fat is metabolically healthier than its mass predicts. Inflammation is itself well documented in lipedema tissue, but its type 2, reparative character suggests that it amplifies this substrate rather than initiating the disease. Against this substrate, however, a focused neuroimmune cluster remains unexplained by purely structural models: a diagnostically accurate quantitative sensory testing pattern, elevated tissue histamine, frequent comorbid fibromyalgia, and disease that is more severe in limbs carrying an additional local vascular trigger. We hypothesize that altered tissue mechanics and adipo-vascular biology help trigger a localized immune cascade, so that these observations may define a clinically identifiable subgroup exhibiting a type 2 immune and mast cell-activation endotype, nested within the broader adipo-vascular disorder. The core framework is deliberately limited to this testable sequence: adipo-vascular susceptibility, per-cell mast cell/type 2 activation, histaminergic peripheral sensitization, the quantitative sensory testing signature, and pain generation in a clinically identifiable endotype. We argue that the endotype is best characterized by per-cell mast cell secretory activity rather than abundance, which is not consistently increased in lipedema tissue. It is defined a priori, before treatment exposure, by a composite of baseline features (affected-fat erythema or warmth, atopic or mast cell-activation features, and the characteristic quantitative sensory testing pattern), with response to mast cell-directed therapy reserved as its prospective test rather than a defining criterion. We present the framework as a graded synthesis, distinguishing evidence shown directly in lipedema from that inferred from related fields and from untested hypotheses. Broader adipogenic, gut-immune, HLA-associated, cancer-related, ADHD-related, and systemic immunomodulatory concepts are separated into a dedicated exploratory section; they are secondary hypotheses and are not required for the central mast cell-histamine-QST-pain claim. The framework identifies mast cell activation as a potential therapeutic target in this subgroup for future investigation, pending prospective validation.

Abstract: Background/Objectives: Chronic spontaneous urticaria (CSU) is a heterogeneous immune-mediated condition involving multiple inflammatory pathways. The clinical utility of platelet-activating factor (PAF) as a biomarker in CSU remains insufficiently defined. This study aimed to evaluate serum PAF levels in patients with CSU and to investigate their associations with clinical characteristics, disease activity, and laboratory and immunological parameters. Methods: This cross-sectional study included 205 adult patients diagnosed with CSU between January 2016 and May 2023. Clinical data, patient-reported outcome measures (PROMs), and extensive laboratory parameters were collected. Serum PAF levels were measured using ELISA in 171 patients. Associations were analyzed using appropriate statistical methods. Results: The cohort was predominantly female (77.6%), with a mean age of 42.6 ± 15.3 years. Most patients had uncontrolled disease (76.9%). The median UAS7 indicated moderate disease activity. PAF levels differed significantly across clinical subgroups. Higher PAF levels were observed in patients with angioedema (alone or in combination with urticaria) compared to those with urticaria alone (p = 0.002), and in patients with predominantly nocturnal and early morning symptoms (p = 0.018). No associations were found between PAF levels and disease activity scores or treatment response. PAF levels showed positive correlations with age, age at symptom onset, IgM, IgG anti-IgE, HSP70, and the CD4/CD8 ratio (all p < 0.05). Conclusions: Circulating PAF levels are associated with specific clinical phenotypes, circadian symptom patterns, and selected immunological markers in CSU, supporting a role for PAF in disease heterogeneity and immune activation. PAF may reflect underlying biological processes not captured by patient-reported disease severity.

Abstract: BCR-mediated antigen internalization is the most efficient pathway for MHC class II presentation to CD4+ T cells, surpassing fluid-phase uptake by up to 10,000‑fold. Using hen egg lysozyme (HEL) complexed with antibodies D1.3 and HyHEL‑10, we show that antibody binding amplifies intrinsic conformational dynamics precisely at the epitope interface. Comparative B‑factor analysis of unbound and antibody‑bound HEL structures, together with NMR order parameters, reveals four findings: (1) antibody epitopes—including the subdominant T cell epitope HEL 112‑129—show elevated B‑factors (+22‑35%) upon complex formation; (2) these regions correspond to pre‑existing dynamic elements (S² = 0.65‑0.72); (3) immune complex size inversely correlates with global HEL B‑factors (global stabilization with local flexibility amplification, termed "focused flexibility"); and (4) HEL 112‑129 serves as a dual B‑cell/T‑cell epitope. Functionally, focused flexibility couples to FcRn‑dependent endosomal sorting: after pH‑dependent dissociation, FcRn binds the antibody Fc and directs recycling, while the conformationally primed antigen is actively trafficked to lysosomal compartments optimized for epitope‑conserving proteolysis. FcRn knockout studies confirm this pathway is required for efficient MHC II loading and CD4+ T cell activation. We propose that antibody binding creates an "optimally unstable" processing intermediate—sterically protected, locally destabilized, and FcRn‑routed—providing a structural and cellular basis for the extraordinary efficiency of BCR‑mediated antigen presentation.

Abstract: Background and Hypothesis: Schizophrenia spectrum disorders (SSD) refer to a collection of conditions causing loss of functionality, disrupted thoughts and perceptions. While the causes of SSD are not fully clear, systemic inflammation has been considered in their pathophysiology. Our purpose was to analyze peripheral inflammation using lymphocyte- and lipoprotein-derived inflammatory ratios in SSD patients. Then it was analyzed the association among these biomarkers, gender, psychopathology, metabolic and clinical data and the possible predictive value to diagnose SSD. Study Design: Cross-sectional case-control study. Blood-derived inflammatory indices were compared between SSD patients (n=351) and healthy control individuals (n=76). Associations with age, gender, body mass index (BMI), and substance use (alcohol, tobacco, xanthine) were evaluated. Diagnostic predictive ability was tested using ROC curve analysis. Study Results: SSD individuals showed significantly higher levels of MLR, SIRI, MHR, LHR, and PHR, along with lower HDL cholesterol and platelet volume, suggesting increased peripheral inflammation. These differences remained significant after accounting for sex and tobacco use. Within the psychosis group, males exhibited higher MHR and LHR values, and tobacco users showed elevated MHR, LHR, and PHR levels. ROC analysis revealed limited diagnostic utility for all tested biomarkers (AUC< 0.7), though findings from previous studies suggest potential value in specific subtypes. Conclusions: Psychotic disorders are associated with subtle yet consistent alterations in inflammation-related blood markers. While demographic and metabolic factors partly influence these indices, their predictive value for psychosis remains limited. These findings support the development of integrative models that combine clinical, social, and biomarkers to improve early identification.

Abstract: Background: The mannan-conjugated birch pollen polymerised allergoid EP-088_T502 has previously been shown to reduce allergic symptoms and anti-allergic medication use during the birch pollen season. This confirmatory phase III trial aimed to evaluate the efficacy, safety, tolerability, and immunologic effects of six pre-seasonal subcutaneous injections.Methods: In this DBPC trial, 278 participants with birch pollen–induced allergic rhinoconjunctivitis (ARC) received either placebo or a cumulative dose of 28,000 mTU EP-088_T502. The primary efficacy endpoint was the combined symptom and medication score (CSMS) during the peak birch pollen season. Safety, tolerability, health-related quality of life (QoL), and immunologic parameters were also assessed.Results: EP-088_T502 significantly reduced the CSMS during the peak birch pollen season compared with placebo (mean absolute difference [MAD] −0.31; p=0.016, FAS). In the PP and Complete Cases sets, MAD were -0.36 (p=0.012) and -0.51 (p=0.013), respectively. Health-related QoL improved by 20% in the EP-088_T502 group compared with placebo during the peak season (p<0.005). EP-088_T502 increased Bet v1-specific IgG4 levels after treatment compared with placebo (6.4-fold; p<0.0001). The Bet v1-specific IgE/IgG4 ratio was reduced by 73% from baseline and by 69% compared with placebo (both p<0.0001). Thirteen systemic allergic reactions occurred, including one grade III systemic reaction considered related to EP-088_T502.Conclusions: EP-088_T502 significantly improved symptom and medication scores in participants with birch pollen–induced ARC and induced a substantial Bet v1-specific IgG4 response. The higher cumulative dose was associated with enhanced immunologic effects and an acceptable safety and tolerability profile.

Abstract: Pediatric autoimmune disorders are increasingly common, but outcomes are poor due to delayed diagnosis. Immune system dysfunctions typically cause these in the presence of essential micronutrient deficiencies, especially vitamin D. Besides, vitamin D deficiency contributes to both immune and endocrine imbalances that may trigger or exacerbate autoimmunity in children. This review highlights the role of micronutrients, cofactors, and mechanisms, especially vitamin D, in regulating immune responses, maintaining hormonal equilibrium, and disease progression. It explores the complex interactions between vitamin D, zinc, selenium, probiotics, and omega-3 fatty acids and the immune and endocrine systems in pediatric autoimmune pathophysiology. The article underscores the importance of early diagnosis, effective interventions, and targeted micro-nutritional strategies to reduce the incidence and severity of these disorders. Proactive supplementation—especially with vitamin D3 and complementary micronutrients—can strengthen immune function, reduce inflammation and oxidative stress, and mitigate the development of childhood autoimmune diseases. Understanding the interconnected roles of different micronutrients, gut microbiota, and immune dysregulation is essential to preventing and effectively managing these conditions. The review encourages pediatricians and other healthcare professionals to adopt integrative approaches, including holistic, orthomolecular, and personalized nutritional therapies, to prevent these disorders and associated complications. Such cost-effective approaches improve clinical outcomes and guide future research and public health strategies. Optimizing micronutrient levels may significantly enhance pediatric well-being and long-term quality of life.

Abstract: Adoptive cellular therapy has emerged as one of the most pivotal modalities in cancer immunotherapy. Although chimeric antigen receptor (CAR)-T cell therapy has achieved remarkable clinical success in hematological malignancies, its broad application is hampered by severe immune-related toxicities, autologous manufacturing limitations, and suboptimal efficacy against most solid tumors, highlighting an urgent unmet clinical need for safer and more versatile cellular therapeutics. As a promising alternative, CAR-NK cell therapy exhibits inherent advantages in safety profile, off-the-shelf accessibility, and multimodal antitumor mechanisms. NKG2D, a pivotal activating receptor broadly expressed on NK cells, specifically recognizes stress-inducible ligands such as MICA/B and ULBP family molecules. These ligands are abundantly overexpressed in various hematological and solid malignancies yet rarely detected in normal tissues, thereby rendering NKG2D an ideal candidate for universal targeted immunotherapy. In this review, we systematically examine the structural design and iterative optimization of NKG2D CAR constructs and elucidate the inherent advantages of CAR-NK over conventional CAR-T cells. We also comprehensively review recent research advances in NKG2D CAR-NK across multiple malignancies, and critically analyze three pivotal translational bottlenecks, including tumor antigen escape, the immunosuppressive tumor microenvironment, and inadequate in vivo persistence. Moreover, we highlight current genetic engineering and combinatorial strategies to overcome these limitations, and outline future research directions focusing on the development of universal off-the-shelf products, multifunctional cell engineering, and rational combination regimens. By integrating mechanistic advances, preclinical findings and early clinical evidence, this review provides a systematic theoretical basis and translational guidance for the structural optimization, clinical translation, and widespread clinical adoption of NKG2D CAR-NK therapy, laying a solid foundation for its future development as a standardized universal anticancer cellular therapeutic.