Exposure to World Trade Center (WTC) dust following the September 11, 2001 attacks has been linked to numerous negative health outcomes in First Responders, including cognitive impairment disproportionately impacting First Responders with aging. However, the mechanisms underlying neurocognitive decline remain poorly characterized. This investigation examined the effects of WTC dust exposure on myelin integrity and neuroinflammatory signaling in twelve-week old male spontaneously hypertensive rats. To model Ground Zero exposure in First Responders, WTC dust was intratracheally administered for two consecutive days (2.43 [± 0.78] mg/day). It was hypothesized that WTC dust exposure would enhance neuroinflammatory signaling and promote an earlier, progressive decline of myelin abundance. Rats were euthanized at 3 and 12 mo post-exposure. Changes in myelin abundance (Black-Gold II), myelin basic protein (MBP) and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase), and microglial responses (ionized calcium-binding protein; Iba1) were assessed through immunohistochemical staining, and cytokines were measured in brain homogenates. WTC dust exposure resulted in increased Black-Gold II staining at 3 mo despite no changes in MBP or CNPase. In contrast, microglial activation and IL-1β concentrations were elevated in dust-exposed rats at 12 mo. Collectively, these data suggest WTC dust exposure induces sustained neuroinflammatory signaling and may elicit a compensatory myelin response indicative of early neurodegeneration.
Atopic dermatitis (AD) is primarily a TH2-driven pathology, yet seasonal factors may activate additional immune pathways. This study examined seasonal variation in plasma cytokines in adults with AD treated with dupilumab (N = 20), AD patients without systemic therapy (N = 15), and healthy controls (N = 34). Cytokines (interleukin [IL]-17A, thymic stromal lymphopoietin (TSLP), interferon (IFN)γ, tumor necrosis factor (TNFα), IL-2, IL-6, IL-12p70, IL-23, and IL-31) were measured during summer and winter under unstimulated and phorbol 12-myristate 13-acetate (PMA)/ionomycin-stimulated conditions. Dupilumab-treated patients showed significant winter increases in stimulated IFNγ, TNFα, IL-2, IL-6, and TSLP, indicating preserved inducible TH1/innate and alarmin activity. Untreated AD patients displayed winter elevation of stimulated IL-6 and unstimulated IFNγ, while healthy controls showed broad winter up-regulation of stimulated TH1/innate cytokines. Higher winter cytokine levels tended to occur in dupilumab-treated patients with scoring atopic dermatitis (SCORAD) > 10 (evaluation of severity AD - scoring of atopic dermatitis). The findings obtained here suggest that seasonal environmental factors modulate immune responses even under biologic therapy, supporting season-aware, individualized AD management.
Bispecific CD3 T-cell engagers (TCE) redirect endogenous T-cells to tumor cells and have demonstrated substantial clinical activity, but their mechanism of action also drives immune-mediated toxicities that are often underpredicted in nonhuman primate (NHP) studies. To investigate the biological basis of this translational gap, we performed a comparative evaluation of human and cynomolgus macaque T-cells using immunophenotyping, functional characterization with TCE, and single-cell transcriptomic profiling. Human and NHP donors showed broadly similar T-cell frequencies, subset distributions, and robust responses to polyclonal stimulation, confirming preserved intrinsic effector capacity in both species. However, across multiple CD20-, BCMA-, and GPRC5D-targeted TCE, human PBMC consistently exhibited greater cytotoxic potency and markedly elevated IFNγ production relative to NHP. Transcriptomic analyses revealed that although global TCR signaling architecture is conserved, human T-cells exhibit increased expression of some proximal signaling mediators, including LAT, PAG1, SKP1, and PI3K-family genes. These functional and molecular differences provide a mechanistic explanation for reduced pharmacologic responsiveness observed in NHP toxicology studies. While NHP studies can provide valuable insight for identifying antigen-dependent cytotoxic hazards, their limitations underscore the increasing importance of incorporating human-relevant models in nonclinical safety assessment.
OBJECTIVE:The aim of our study is to evaluate the laboratory results of sensitization to molecular components in atopic dermatitis patients and to show if there is the significant relation between the sensitization to molecular components of food allergens (regardless of the occurrence of clinical reactions) and the sensitization to molecular components of inhalant allergens. MATERIALS AND METHODS:The complete dermatological and allergological examination including the examination with Multiplex ISAC testing was performed in all included patients. This study analyzed the data to determine the relation between the sensitization to molecular components of food and inhalant allergens. This study used the statistical calculation with matrix. RESULTS, DISCUSSION, AND CONCLUSION:Altogether 100 atopic dermatitis patients were examined - 48 men, 52 women, the average age 40.9 years. The significant relation between the sensitization to molecular components of timothy and molecular components of food allergens (PR 10 proteins, storage proteins, and lipid transfer proteins) was confirmed. Also, the significant relation between the sensitization to PR 10 proteins of food allergens and sensitization to Aln g 1, Bet v 1 (PR 10 proteins), Ole e 9 (Beta-1,3-glucanase), and Bet v 2, Mer a 1 (Profilin) was recorded.
The fire ant Solenopsis invicta is an aggressive invasive species whose venom frequently triggers hypersensitivity reactions, including severe anaphylaxis. In endemic regions, its stings represent a significant cause of Hymenoptera-related allergy. Four venom allergens have been identified - phospholipase A1 (Sol i 1), antigen 2 (Sol i 2), antigen 3 (Sol i 3), and antigen 4 (Sol i 4) - with Sol i 3 recognized as the predominant sensitizer. However, the molecular determinants that drive Sol i 3 allergenicity and its potential cross-reactivity with other Hymenoptera venoms remain insufficiently understood. This study identified the linear immunoglobulin E (IgE) epitopes of Sol i 3 and examined their recognition by sera from yellow jacket venom (YJV) - and Polistes wasp-allergic patients. Two linear epitopes were mapped: Sol i 3_e1 (ELRQRVASGKEMRG) and Sol i 3_e2 (WAKTTKIGCGRIMF). Although Sol i 3 exhibits limited sequence and structural similarity to other antigen 5 proteins, it contains a conserved immunoreactive core (WAKTTK), analogous to the WAKTKE motif described for the allergen Poly p 5 from Polybia paulista. This conserved region may represent a shared epitope contributing to cross-reactivity among Hymenoptera venoms. Consistently, sera from P. dominula-sensitized patients and YJV-sensitized patients recognized Sol i 3_e2. These findings define key B-cell epitopes of Sol i 3 and reveal a conserved motif that may underlie cross-reactivity, offering implications for improved diagnosis and immunotherapy.
Neutrophil functions are tightly regulated by numerous membrane receptors, the activation of which determines the appropriate response to various endogenous and exogenous factors. Accumulating evidence indicates that environmental endocrine-disrupting chemicals (EDC) can modulate immune system function, inter alia, by influencing neutrophil activity. Imazalil (IMZ), an EDC known to exhibit antagonistic activity against the androgen receptor expressed on neutrophils, exemplifies a compound that may potentially affect these processes. The aim of this study was to evaluate the effect of imazalil and testosterone on the expression of CD molecules associated with fundamental neutrophil functions. Whole blood collected from healthy men served as the study material. Plasma imazalil concentrations were determined using GC-MS. Isolated neutrophils were incubated in the presence of IMZ (at a concentration established in the present study: 28 ng/ml) or testosterone (at a physiological concentration). Cytometric analysis revealed that exposing cells to imazalil resulted in an elevated percentage of neutrophils expressing CD14, CD66b, CD62L, and CD284, accompanied by a decrease in the percentage of cells expressing CD10, CD11c, CD15, CD49d HLA-DR, as well as a reduction in MPO+ neutrophils. No changes in the percentage of neutrophils expressing the analyzed molecules were observed in the presence of testosterone. In conclusion, imazalil modulates the neutrophil immunophenotype, which may inhibit functions such as adhesion, migration, chemotaxis, and phagocytosis. Consequently, this may predispose individuals exposed to this compound to innate immunity disorders.
Fine particulate matter (PM2.5) exposure contributes to over 4 million premature deaths annually, yet the mechanistic role of lung microbiota in PM2.5-induced pulmonary inflammation remains poorly understood. In collaboration of 16S rRNA and single-cell RNA multi-omics analysis and in vivo/in vitro experimental validation with antibiotic intervention strategies, the study here examined PM2.5-microbiota interactions in murine PM2.5 exposure models and cellular systems. It was found that PM2.5 exposure induced lung microbiota dysbiosis characterized by Gram-negative bacterial expansion, particularly Proteobacteria dominance, accompanied by reduced microbial diversity. scRNA analysis revealed coordinated activation of TLR4/MyD88/NLRP3 inflammatory signaling pathways and p53/p21/p16-mediated cell cycle arrest. Moreover, PM2.5 exposure activated NLRP3 inflammosome-dependent macrophage pyroptosis as evidenced by increased interleukin (IL)-1β, IL-18, caspase-1, and GSDMD expression. In vitro studies demonstrated that the inflammatory changes induced by PM2.5 exposure were statistically indistinguishable from those of LPS-positive controls, confirming endotoxin-like mechanisms. Critically, antibiotic pretreatment effectively attenuated PM2.5-induced inflammatory responses, cell cycle arrest, and tissue pathology, which established causality between microbiota disruption and pulmonary dysfunction. In conclusion, this study revealed lung microbiota dysbiosis as a critical mediator of PM2.5-induced pulmonary inflammation through Gram-negative bacterial expansion and subsequent endotoxin-like activation of inflammatory cascades, thereby providing novel mechanistic insights and potential microbiome-targeted therapeutic strategies for air pollution-associated respiratory diseases.
Drug-induced autoimmune (DIA) toxicity represents a significant challenge in drug safety evaluation, and traditional autoantibody biomarker antinuclear antibodies (ANA) exhibits limitations in DIA assessment, highlighting the urgent need for more reliable biomarkers. This study employed d-penicillamine to induce an autoimmune disease model in Brown Norway (BN) rats and systematically investigated the expression changes of autoimmunity-related microRNAs (miRNAs) in spleen, exploring their potential value as novel biomarkers for predicting DIA toxicity. A total of 30 male BN rats were randomly divided into control group (0 mg/kg), low-dose d-penicillamine group (150 mg/kg), and high-dose group (450 mg/kg), with necropsy performed at days 7 and 14 post-administration. ANA detection, histopathological examination of spleen, surface immune-related proteins CD11a and CD278 on splenic CD3+CD4+ T-cells, and miRNA expression profiling in spleen tissue were conducted. Receiver operating characteristic (ROC) curves were plotted to evaluate the correlation of ANA and miRNAs with the degree of splenic pathological damage, and the relationships between miRNAs and both ANA and CD11a/CD278 were analyzed. The results demonstrated that 16 miRNAs were significantly up-regulated in spleen tissue, among which 13 miRNAs showed superior predictive performance (AUC = 0.828-0.898) compared to the traditional biomarker ANA (AUC = 0.783), with most of these miRNAs exhibiting correlation with CD11a/CD278 expression. These findings indicate that aberrant miRNA expression in the d-penicillamine-induced autoimmune disease preclinical model in BN rats holds potential for early prediction of DIA toxicity, with some miRNAs demonstrating higher diagnostic value than ANA, thereby providing novel molecular biomarker candidates for early diagnosis of DIA toxicity
Community-acquired pneumonia (CAP) remains a major cause of morbidity and mortality, particularly in older adults and patients with chronic co-morbidities. Cytokine patterns and simple hematological indices may improve risk stratification beyond conventional clinical scores. In this prospective single-center study, 41 adults with CAP treated at Kaunas Hospital of Lithuanian University of Health Sciences between November 2024 and March 2025 were enrolled. Clinical data, pneumonia severity index (PSI), complete blood count-derived indices (neutrophil-lymphocyte ratio [NLR], platelet-lymphocyte ratio [PLR], systemic immune-inflammation index [SII]), and serum concentrations of interleukin (IL)-6, IL-8, interferon (IFN)-γ, and G-CSF were obtained on admission (Visit 1) and after 7 days of treatment (Visit 2). Patients were stratified by age (≤65 vs >65 years), co-morbidities, and PSI class. Non-parametric tests and Spearman correlations were applied. The results indicate that patients with co-morbidities and those > 65 years had significantly higher IL-6 levels than younger and non-comorbid patients. IL-6, IFNγ, and G-CSF concentrations were highest at admission and declined by day 7, particularly in PSI Class II patients. Higher PSI classes were associated with increased IL-8, IL-6, and IFNγ. NLR and SII were significantly higher in older patients and in PSI III-IV compared with PSI I-II. IL-6 and G-CSF showed strong positive correlations with NLR and SII, especially in elderly and comorbid patients, whereas PLR displayed weaker and less consistent associations. From these data, it is concluded that in CAP, serum IL-6, IFNγ, and G-CSF reflect age, co-morbidity burden, and disease severity, while NLR and SII closely mirror cytokine-driven systemic inflammation. These readily available indices may serve as cost-effective prognostic markers and, combined with cytokine profiling, could enhance early risk stratification and guide individualized management in CAP.
Atopic dermatitis (AD) is a chronic inflammatory skin disorder characterized by impaired skin barrier function, persistent inflammation, and increased vulnerability to environmental irritants. Chronic inflammation in AD can also contribute to premature skin aging by promoting collagen degradation, wrinkle formation, and reduced skin elasticity. Conventional treatments, including topical corticosteroids, may lead to adverse effects with long-term use, underscoring the need for safer, sustainable alternatives. Glycomacropeptide (GMP), a bioactive peptide derived from κ-casein during cheese production, has attracted interest for its anti-inflammatory, antioxidant, and skin barrier-supporting properties. Evidence from in vitro, in vivo, and limited clinical studies suggests that GMP may modulate inflammatory responses, attenuate oxidative stress, support collagen integrity, and promote wound healing. Its amphiphilic nature also enables its use as a natural emulsifier in topical formulations. In addition, the utilization of GMP contributes to sustainable development by valorizing dairy byproducts. This narrative review synthesizes current literature on GMP's dermatological potential, with a focus on its applicability in AD management and skin aging mitigation, while identifying research gaps and directions for future clinical evaluation.
This review was undertaken because it is known that high doses of atrazine activate the HPA axis, leading to increased corticosterone secretion, which could result in immunosuppression. The mammalian immunotoxicity of atrazine was evaluated based on in vitro and in vivo studies and the association between chlorotriazine use and the risk of cancer in humans based on epidemiological studies. In vitro studies reported that μM concentrations of atrazine may adversely affect cytokine production, lymphocytes, and dendritic cell function. However, there were no consistent patterns of effects of the chlorotriazines on the developing or mature immune system in sub-chronic, chronic, and lifetime animal studies. In a detailed immunotoxicity study on atrazine in rats, there were no effects of atrazine treatment on T-cell-dependent, IgM antibody production or natural killer cell activity. Based on the compendium of toxicology data reviewed, it was concluded that atrazine is unlikely to be immunotoxic at any dose to which humans might realistically be exposed. This conclusion was supported by epidemiological studies indicating that there was no consistent association between occupational exposure to atrazine and cancers of the immune system.
Exposure to environmental agents has been linked to inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn’s disease (CD) in humans. In these studies, we describe a modification of an experimental model used historically in the pharmaceutical industry to help identify therapies for the treatment of IBD to facilitate its use for identification of environmental agents that have the potential to accelerate, exacerbate, and/or impair recovery from IBD. In this model, female C57BL/6 mice were exposed to low levels of dextran sodium sulfate (DSS) for 7 consecutive days in drinking water to allow for a modest level of colon inflammation and pathology as measured by a battery of clinical, pathological, toxicological endpoints (water consumption, body weights, colon length, body temperature, stool consistency, and hematochezia), and cytokine/chemokine production in the serum and colon. Treatment with DSS for 7 d showed a clear dose-response with 1% DSS producing minimal changes in the colon and 3% DSS inducing severe damage with IBD. A concentration of 2% DSS in drinking water for 7 d was selected for investigating disease recovery and exacerbation by an environmental agent as it induced mild colon inflammation that showed nearly complete resolution within 21 d following cessation of DSS exposure. Cytokine and chemokine profiles showed a Type 1 predominant immune response in the colon and serum that is consistent with inflammation observed in human IBD. The model was used to determine the impact of administration of a high salt diet (HSD) on DSS IBD progression, severity, and recovery. While administration of HSD by itself had no effect on indicators of colon damage or inflammation, co-administration of HSD with DSS, produced marked exacerbation and persistence of disease supporting the potential of the model for identifying environmental agents that can affect IBD.
This study investigated the relationship between CD200 molecule expression on B- lymphocytes and the levels of specific IgE to molecular components of storage mites (Gly d 2, Lep d 2), dog (Can f 1, Can f 2), cat (Fel d 1), shrimp (Pen m 2), molds (Asp f 6, Mala s 11, Alt a 6, Alt a 1, Mala s 6, Cla h), and German cockroach (Bla g 9) in patients with atopic dermatitis (AD), both with and without dupilumab therapy. The study included 46 patients with AD- 26 without dupilumab treatment and 20 with dupilumab treatment. Serum levels of specific IgE were measured using the ALEX2 Allergy Xplorer diagnostic microarray and CD200 molecule was evaluated with the use of flow cytometry. Spearman's rank correlation coefficient was used to assess the relationship between B-lymphocyte CD200 expression and specific IgE levels to molecular components. According to the results, the association between CD200 expression and specific IgE levels to molecular components was low (up to 10%) in AD patients without dupilumab therapy. In patients with dupilumab therapy, the association was non-linear, indicating that the two monitored parameters had opposite effects. In conclusion, the present study did not confirm any association between the CD200 molecule on B-lymphocytes and specific IgE levels to molecular components.
The aim of the current study was to evaluate the incidence of soy allergy in patients with atopic dermatitis (AD) and to evaluate the results of specific IgE against molecular components of soy. Altogether, 100 AD patients were examined. Soy allergy was confirmed in an open exposure test (history), and the presence of specific IgE against molecular components of soy (Gly m 4, Gly m 5, Gly m 6, Gly m 8) was evaluated using an ALEX2 Allergy Explorer test. The results for the measures of specific IgE against molecular components of soy (Gly m 4, Gly m 5, Gly m 6, Gly m 8) and clinical reactions in the open exposure test were then compared. Soy allergy was confirmed in 12% of patients. The sensitivity of specific IgE against Gly m 4 was 50.0% (21.1-78.9%). In another 29% of patients we recorded the positive results for specific IgE against Gly m 4 without any clinical reaction to soy. Compared to results from a previous study in 2013, there was an increase in the incidence of soy allergy in AD patients. An elimination diet and an exposure test are recommended to detect a reaction to soy. ALEX2 Allergy Xplorer test gives us a comprehensive picture of sensitization and the possibility of evaluation of cross-reacting allergens.
Unwanted immunogenicity of therapeutic proteins arises through the combination of many factors, with the route of administration considered a significant contributor. Contrary to historic data on vaccine delivery, analysis of various therapeutic protein products indicates that the subcutaneous route is not a systematic risk. However, individual product assessments may identify factors specific to the circumstance of their use. Preclinical in vivo studies may add additional information to the comparative immunogenicity risk assessment of intravenous versus subcutaneous administrations. Moreover, immunogenicity risk assessment of new biotherapeutic modalities, such as bispecific antibodies and antibody-linked cytokines, may benefit from a full analysis of risk factors, including preclinical in vivo data. The study here provides immunogenicity analysis of an IgG, two CD3 bispecific antibodies, and two Fc-linked immunocytokines administered intravenously and subcutaneously, aiming to highlight similarities and differences between these administration routes. The current results suggest that the development of anti-drug antibodies does not solely depend on the route of administration but is influenced by multiple risk factors, which should be addressed on a case-by-case basis. This paper reflects on the challenges of interpreting the data and propose standards for improving sample and data collection to aid future analysis.
The aim of this study was to assess the relationship between the expression of the CD23 molecule on B-cells and the levels of specific IgE against allergens and molecular components of storage mites (Gly d 2, Lep d 2), dog (Can f 1, Can f 2), cat (Fel d 1), shrimp (Pen m 2), molds (Asp f 6, Mala s 11, Alt a 6, Alt a 1, Mala s 6, Cla h), and German cockroach (Bla g 9) in atopic dermatitis (AD) patients (with and without dupilumab therapy). Here, 46 patients with AD were included (26 without dupilumab treatment, 20 with dupilumab treatment). Serum levels of specific IgE were measured using the component-resolved diagnostic microarray ALEX2 Allergy Xplorer, and the expression of the CD23 molecule on B-cells was evaluated using flow cytometry. For statistical analysis, a Spearman’s rank correlation was used. The data indicated there was a higher correlation between CD23 expression on B-cells and specific IgE against molecular components of storage mites Bla g 9 (up to 27%), cat Fel d 1 (22.7%), and allergen extract Cla h (Cladosporium herbarum) up to 38.9% in AD patients treated with dupilumab. These results regarding the higher association suggested a significant role in the non-inflammatory clearance and uptake of these specific IgE antibodies.
Trichloroethylene (TCE) is a volatile synthetic chemical used in various industrial processes like metal degreasing. Large amounts of TCE have been released into the environment. Exposure to TCE can occur through routes, such as inhalation for workers using TCE or ingestion of drinking water in contaminated areas. Macrophages are key immune cells in virtually all tissues in the human body, including the fetal membranes, making them a plausible target for DCVC-induced immunotoxicity. Macrophages are critical for maintaining anti-microbial defenses during pregnancy, but little data exists on TCE immunotoxicity during pregnancy. We previously showed that the TCE metabolite, S-(1,2-dichlorovinyl)-L-cysteine (DCVC), down-regulates immune functions in fetal membranes. To gain insight into immune functions impacted by DCVC, we treated a macrophage cell model (THP-1 cells) with DCVC followed by stimulation with bacterial or fungal toxins relevant for intrauterine infections: lipopolysaccharide (LPS), lipoteichoic acid (LTA), or zymosan. DCVC inhibited toxin-stimulated release of cytokines (e.g. TNFα and IL-1β) for all three microbial toxins. We then conducted benchmark dose modeling and compared benchmark doses for DCVC cytotoxicity vs. cytokine suppression and determined that inhibition of cytokine release was the more potent endpoint compared to cytotoxicity. Finally, we analyzed a previously generated transcriptomic dataset from THP-1 cells stimulated with LPS, with or without DCVC treatment. We identified transcription factors that were enriched with DCVC and/or LPS treatment, including NF-kB and Vitamin D receptor (VDR). Our findings show that DCVC potently alters cellular and molecular macrophage immune responses involved in defense against intrauterine pathogens.
The aim of the study here was to evaluate the association between expression of CD23 molecule on B-lymphocytes and the level of specific IgE to molecular components of birch, Bermuda grass, hazel pollen, timothy, and rye grass in atopic dermatitis (AD) patients (with and without dupilumab therapy). A total of 46 patients suffering from AD were included: 26 without dupilumab treatment and 20 with dupilumab treatment. Serum levels of specific IgE were measured by the components resolved diagnostic assay ALEX2 Allergy Xplorer, the expression of CD23 molecule on B-lymphocytes was evaluated with flow cytometry. For the statistical analysis, the Spearman’s rank correlation coefficient was used. In patients treated with dupilumab, the higher association was observed between the expression of CD23 on B-lymphocytes and specific IgE to molecular components Bet v 1, Cor a 1.0103, Cor a 1.0401, and Phl p 1. This study demonstrated that the relationship between CD23 expression on B-lymphocytes and specific IgE to pollen molecular components varies depending on whether the patient was treated with dupilumab and the type of molecular component involved.
In December 2019, the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was identified in Wuhan, China, leading to the global Coronavirus Disease pandemic. The rapid spread of SARS-CoV-2 highlighted the urgent need for effective vaccines. However, the high cost, cold storage requirements, and scalability challenges associated with mRNA vaccines have necessitated alternative vaccine technologies. In the study, the safety of a plant-based vaccine was evaluated. The vaccine, an emulsion of the SARS-CoV-2 S1 antigen and a synthetic TLR4 agonist produced and purified from Nicotiana benthamiana, was administered to Sprague-Dawley rats three times over 4 wk. Mortality, clinical signs, body weight, food consumption, vision, urinalysis, gross findings, organ weight, hematology, serum biochemistry, histopathology, and immunogenicity were evaluated. The results showed that antibodies were efficiently produced and maintained for one month following vaccination with the plant-derived receptor-binding domain (RBD) antigen of COVID-19. Furthermore, the rats showed no toxicological symptoms, with reversible changes at the injection site and minor histological alterations in the spinal cord and bone marrow, typical of vaccine responses. The plant-derived SARS-CoV-2 vaccine appears safe following repeated administration over 4 wk and represents a promising alternative for potential use in human clinical trials and clinical applications.
