Journal of Immunotoxicology最新文献

It was previously reported that half of the anaphylaxis cases occurring after intra-articular administration of diclofenac etalhyaluronate (DEH) - developed as SI-613/ONO-5704 and marketed as JOYCLU^® - were induced by IgE-mediated mechanisms; mechanisms for the remaining cases remain unclear. In this study, we investigated the relationship of DEH-induced anaphylaxis to non-IgE-mediated mechanisms in vitro. Assays were carried out based on the production of downstream products of the complement cascade, calcium influx due to Mas-related G protein-coupled receptor-X2 (MRGPRX2) activation, mast cell degranulation, and expression of basophil activation markers. Human plasma, CHO-K1 cells stably expressing MRGPRX2, the human mast cell line LAD2, and the human basophil leukemia cell line KU812 were used for these evaluations. No effect of DEH treatment was found on complement activation, MRGPRX2 agonist activity, direct mast cell activation, or direct basophil activation. From this it could be concluded that DEH-induced anaphylaxis is unlikely to involve complement activation or direct activation of mast cells and basophils. However, the possibility remains that the anaphylaxis might be a non-immunological hypersensitivity reaction due to inhibition of cyclooxygenase-1 by non-steroidal anti-inflammatory drugs (NSAID). Further investigation into the relationship between the non-immunological hypersensitivity and anaphylaxis following DEH administration is warranted.

Plantago asiatica L., a perennial herb in the family Plantaginaceae, has been shown to impart several pharmacologic activities, including anti-oxidative, anti-inflammatory, and diuretic effects. In the study here, the anti-gout(y) arthritis (GA) effects of a crude extract from P. asiatica L. (PAE) were investigated in a rat GA model. For this, PAE was prepared by ethanol extraction and analyzed for phytochemicals by RP-HPLC and Q-TOF-MS. Thereafter, potential therapeutic effects of the PAE were investigated in rats; Wistar rats (male, 8 wk-of-age) were randomly allocated into four groups (n = 9/group) and intra-articularly injected with 3 mg monosodium urate (MSU) in saline solution to establish a GA model. For the study, rats received oral dosings of 0.3 mg colchicine/kg or 1 g PAE/kg (w/w) before and after gout was established. At fixed times after the treatments, assessment of joint swelling ratios and pathological changes in the joints, as well as of select cytokine expression in the blood, was done. RP-HPLC results showed the PAE contained at least 8 'active' ingredients, with plantamajoside, verbascoside, and cymaroside being the most abundant. In comparison to in control rats, MSU induced joint space narrowing, ankle joint swelling, and increased levels of pro-inflammatory interleukin (IL)-1β, IL-17a, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ, and reductions in anti-inflammatory IL-10 in the blood. PAE treatment significantly reversed patho- genic joint space narrowing and swelling, reversed the MSU-induced changes in inflammatory factors, and in general imparted effects very similar to those seen with colchicine (COL; known non-steroidal anti-inflammatory drug for clinical treatment of GA). Collectively, these findings provide experimental evidence supporting the potential applicability of PAE to treat gouty arthritis.

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.

Efgartigimod is a human IgG₁ antibody F_c fragment that reduces IgG levels through neonatal F_c receptor blockade. This study evaluated whether efgartigimod affects the generation of T-cell-dependent antibodies and cellular immune responses to keyhole limpet hemocyanin (KLH) immunization in non-human primates. Cynomolgus monkeys received efgartigimod or vehicle control intravenously for 11 wk, followed by a recovery phase. KLH challenges occurred during both the dosing phase and the recovery phase. No statistically significant differences emerged in anti-KLH IgM levels between the efgartigimod and control groups. Likewise, comparable KLH-specific T cell responses were observed between groups. Anti-KLH IgG titers were lower in efgartigimod-treated animals compared with controls only after the first boost of KLH, coinciding with decreases in total IgG titers in efgartigimod-treated animals, and returned to baseline levels by the end of the recovery phase. Taken together, these results indicate that efgartigimod does not suppress T-cell-dependent antibody responses or antibody class-switching. The findings of this study are consistent with efgartigimod's pharmacological mechanism of action and suggest that efgartigimod does not impair the generation of effective immune responses.

Inflammation plays a critical role in prostate cancer (PCa) pathophysiology, yet the diagnostic value of specific inflammatory markers remains unclear. This study evaluates the association between circulating and tissue inflammatory markers with PCa presence and their potential as biomarkers for risk stratification. This prospective study analyzed serum and prostate biopsy samples from 60 patients with PCa and 22 cancer-free controls. Concentrations of inflammatory markers, including IL-2, IL-4, IL-10, IL-13, IL-33, Oncostatin M, TNFα, PDGF-BB, and TREM-1, were measured using Luminex technology. Statistical analyses included the Mann-Whitney test, logistic regression, and ROC curve analysis to assess differences and diagnostic performance. PCa patients exhibited significantly higher serum levels of IL-2 (p = 0.001), IL-10 (p < 0.001), IL-33 (p < 0.001), Oncostatin M (p = 0.018), and TNFα (p = 0.017) compared to controls. In contrast, biopsy tissue levels of IL-4 (p < 0.001), IL-10 (p < 0.001), IL-13 (p = 0.004), Oncostatin M (p = 0.012), PDGF-BB (p = 0.039), and TREM-1 (p = 0.013) were significantly lower in PCa patients, suggesting an inverse association. IL-10 (inverse) and IL-4 (inverse) in biopsy tissue showed high specificity in ROC analysis (AUC = 0.788 and 0.804, respectively), while IL-2 and IL-33 in serum were positively associated with PCa risk. This study suggests that IL-4, IL-10, and IL-13 in biopsy tissue may serve as biomarkers of a protective effect, while elevated IL-2 and IL-33 in serum are associated with an increased risk of PCa. These findings highlight the potential of inflammatory markers in PCa risk stratification, warranting further investigation in larger cohorts.

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.

Cell-mediated immune (CMI) responses to adeno-associated virus (AAV) can lead to tissue damage and loss of therapeutic transgene expression. Identifying robust biomarkers and mechanisms of CMI can aid clinical practice and advancement of AAV gene therapies. The present work evaluated peripheral blood mononuclear cells (PBMC) from non-human primates (NHP) before and after immunization with adenovirus 5 encoding AAV9 capsid antigen. PBMC were stimulated ex vivo with AAV9 capsid peptides to evaluate CMI responses by interferon (IFN)-γ ELISpot, intracellular cytokines/activation markers, secreted cytokines, and RNAseq. AAV peptide stimulation produced a robust IFNγ ELISpot 11 days after immunization and ≈ 4 years after cryopreservation. Flow cytometry revealed increased IFNγ, interleukin (IL)-2, or tumor necrosis factor (TNF)-positive T-cells. Increases in secreted CXCR3 ligands (IP-10, I-TAC) were detected. Robust changes and correlations to ELISpot responses were revealed by RNAseq, including IFNγ, IP-10, and I-TAC, many downstream transcripts, and several IFN-independent pathways. These data from AAV-immunized NHP identify biomarkers that could serve as robust and sensitive supplements/alternatives to ELISpot for early detection of CMI responses. Assessment of these biomarkers in non-clinical and clinical studies is a critical next step to determine the translation of this work to administration of a therapeutic AAV vector.

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.

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.