Search Results (817)

Pollen–food allergy syndrome (PFAS), also known as oral allergy syndrome, is a common condition affecting individuals sensitized to pollens such as birch, ragweed, and grass. This syndrome arises from immunological cross-reactivity between pollen allergens and structurally similar proteins found in various fruits, vegetables, and nuts. Although typically presenting with mild oral and pharyngeal symptoms, PFAS can occasionally result in severe allergic reactions, underscoring its clinical significance. This review explores the pathophysiology of PFAS, highlighting the molecular mechanisms underlying cross-reactivity and examining the main protein families involved, including those contributing to variations in symptom severity. Current diagnostic approaches, including skin prick testing, specific immunoglobulin E measurements, and component-resolved diagnostics, are discussed. Emerging diagnostic tools and biomarkers with potential to enhance accuracy are also examined. Therapeutic strategies for PFAS primarily focus on symptom management and avoidance of trigger foods. However, novel approaches such as allergen immunotherapy and biologics targeting key immune pathways are gaining traction as potential interventions for more severe or refractory cases. By addressing the diagnostic and therapeutic challenges of PFAS, this paper aims to provide clinicians and researchers with a comprehensive understanding of this condition, fostering improved patient care and the development of innovative treatment strategies. Full article

Endothelial dysfunction is a hallmark of several pathological conditions, including cancer, cardiovascular disease and inflammatory disorders. In these conditions, perturbed TCA cycle and subsequent succinate accumulation have been reported. The role of succinate as a regulator of immunological responses and inflammation is increasingly being recognized. Nevertheless, how endothelial cell function and phenotype are altered by elevated intracellular succinate has not been addressed yet. Thus, we employed numerous in vitro functional assays using primary HUVECs and diethyl succinate (DES), a cell membrane-permeable succinate analogue. An MTS assay 1 h post stimulation with DES suggested reduced metabolic activity in HUVECs. Concurrently, elevated production of ROS, including mitochondrial superoxide, and a reduction in mitochondrial membrane potential were observed. These findings were corroborated by Seahorse mito-stress testing, which revealed that DES acutely lowered the OCR, maximal respiration and ATP production. Given the link between mitochondrial stress and apoptosis, we examined important survival signalling pathways. DES transiently reduced ERK1/2 phosphorylation, a response that was followed by a skewed pro-apoptotic shift in the BAX to BCL2L1 gene expression ratio, which coincided with upregulating VEGF gene expression. This indicated an induction of mixed pro-apoptotic and pro-survival signals in the cell. However, the BAX/BCL-XL protein ratio was unchanged, suggesting that the cells did not commit themselves to apoptosis. An MTS assay, caspase 3/7 activity assay and annexin V/propidium iodide staining confirmed this finding. By contrast, stimulation with DES induced acute endothelial barrier permeability, forming intercellular gaps, altering cell size and associated actin filaments without affecting cell count. Notably, during overnight DES exposure gradual recovery of the endothelial barrier and cell sprouting was observed, alongside mitochondrial membrane potential restoration, albeit with sustained ROS production. COX-2 inhibition and EP4 receptor blockade hindered barrier restoration, implicating a role of COX-2/PGE₂/EP4 signalling in this process. Interestingly, ascorbic acid pre-treatment prevented DES-induced acute barrier disruption independently from ROS modulation. In conclusion, succinate acts as a significant regulator of endothelial mitochondrial function and barrier integrity, a response that is counterbalanced by upregulated VEGF and prostaglandin production by the endothelial cells. Full article

Non-specific Lipid Transfer proteins (nsLTPs) are relevant allergens of several pollens and plant foods. Sensitization to nsLTPs is not typical in our region. Still, it has become an increasingly common cause of IgE-mediated food allergies and food-induced anaphylaxis in Northern Europe in recent decades. No in-depth studies describe the prevalence of sensitization of molecular components to nsLTPs in Lithuania. This study aimed to determine the sensitization profile of atopic patients at the Immunology and Allergy Department of Kauno Klinikos to the components of nsLTPs, using molecular allergen component analysis. Sixty Lithuanian adults with symptoms of allergic rhinitis and/or allergic asthma and/or food allergies were included into the study. Specific immunoglobulin E (IgE) levels were measured using two in vitro techniques: allergen extract and molecular component analysis. Results showed that 25% of subjects were sensitized to nsLTP-containing allergen sources, mostly to Zea m 14, Mal d 3, Vit v 1, and Art v 3. The median amount of total IgE was higher in nsLTP-sensitized patients than in nsLTP-nonsensitized patients. Based on Cohen’s Kappa and McNemar tests, the results of allergen extract and component analysis tests do not always agree, especially when we determine the sensitization to allergen sources containing nsLTPs. Molecular allergen component analysis could be the first choice in determining detailed sensitization to nsLTPs in patients who experienced anaphylaxis of unknown origin. Full article

There is a high medical need to develop new strategies for the treatment of patients with acute myeloid leukemia (AML) refractory to conventional therapy. In vitro, the combinations of the blast-modulatory response modifiers GM-CSF + Prostaglandin E1, (summarized as Kit M) have been shown to convert myeloid leukemic blasts into antigen-presenting dendritic cells of leukemic origin (DC_leu) that were able to (re-)activate the innate and adaptive immune system, direct it specifically against leukemic blasts, and induce memory cells. This study aimed to investigate the immune modulatory capacity and antileukemic efficacy of Kit M in vivo. Brown Norway rats suffering from AML were treated with Kit M (twofold application). Blasts and immune cells were monitored in peripheral blood (PB) and spleen. Upon the observation of promising immune modulatory effects in the treated animals, two patients with AML refractory to multiple lines of therapy were offered treatment with Kit M on an individualized basis. Safety, as well as immunological and clinical effects, were monitored. Samples obtained from a third patient in similar clinical conditions not receiving Kit M were used as controls for immune monitoring tests. Animal experiments: Drugs were well tolerated by the treated animals. After 9 days of treatment, DC_leu and memory-like T cells increased in the peripheral blood, whereas regulatory T cells, especially blasts, decreased in treated as compared to untreated control animals. Clinical courses: No severe side effects were observed. In patient 1482, PB blasts remained under the detection threshold during 27 days of treatment, thrombocytes were normalized, and (leukemia specific) immune effector cells of the adaptive and innate immune system increased up to 800-fold compared to the start of treatment. Patient 1601 responded with a 12% reduction in blasts in PB immediately after Kit M treatment. Several subtypes of (leukemia-specific) immune effector cells in PB increased up to four-fold during the 19 days of treatment. In contrast, immune-reactive cells decreased under mild chemotherapy in the PB of control patient 1511 with comparably refractory AML. Within the limitation of low numbers in both animal experiments and clinical applications, our data suggest that Kit M treatment of AML-diseased rats and patients is feasible and may induce leukemia-specific immune reactions and clinical improvement. A larger series and a prospective clinical trial will be required to confirm our observations. Beyond optimized doses and schedules of the applied compounds, the combination with other antileukemic strategies or the application of Kit M in less proliferative stages of the myeloid diseases need to be discussed. If effects are confirmed, the concept may add to the armamentarium of treatments for highly aggressive blood cancer. Full article

Background: The inflammation associated with overweight and obesity seems to alter iron metabolism, but there are few studies evaluating those conditions in children. Thus, we aimed to evaluate the leukometric, immunological, and hematimetric parameters of overweight and obese schoolchildren. Methods: This is a cross-sectional study in which 39 children living in Chonim de Cima (Brazil) underwent anthropometric, hematological, and immunological assessments. The evaluated parameters were compared between the study group (overweight/obesity, n = 15) and the control group (n = 24). Unpaired t-test, Mann–Whitney test, and linear regression were used for statistical tests, and the panoramic profile was used to illustrate differences between groups. Results: The study group had lower mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) and higher TNF levels compared to the control group. Positive correlations were observed between BMI-for-age percentile and total leukocytes (r = 0.1493; p = 0.0151) or neutrophils (r = 0.1395; p = 0.0192). Negative correlations between the BMI-for-age percentile and MCV (r = 0.1464; p = 0.0162) and MCH (r = 0.1460; p = 0.0164) were found. Furthermore, through the panoramic profile, it was noted that the study group had a higher frequency of individuals with high levels of TNF and lower frequencies of individuals with increased hemoglobin and serum iron. Conclusions: Our data suggest that overweight and obesity contribute to a pro-inflammatory context (leukocytes, neutrophils, and TNF) and MCV and MCH reduction in schoolchildren. Full article

Immunization against Gonadotropin-Releasing Hormone (GnRH) has been successfully explored and developed for the parenteral inoculation of animals, aimed at controlling fertility, reducing male aggressiveness, and preventing boar taint. Although effective, these vaccines may cause adverse reactions at the injection site, including immunosuppression and inflammation, as well as the involvement of laborious and time-consuming procedures. Oral vaccines represent an advancement in antigen delivery technology in the vaccine industry. In this study, a Salmonella enterica serovar Typhimurium (S. Typhimurium) mutant lacking the pathogenicity island 2 (S. Typhimurium ΔSPI2) was used as a vehicle and mucosal adjuvant to deliver two genetic constructs in an attempt to develop an oral immunological preparation against gonadotropin hormone-releasing hormone (GnRH). S. Typhimurium ΔSPI2 was transformed to carry two plasmids containing a modified GnRH gene repeated in tandem (GnRXG/Q), one under eukaryotic expression control (pDNA::GnRXG/Q) and another under prokaryotic expression control (pJexpress::GnRXG/Q). A group of three male BALB/c mice were orally immunized and vaccination-boosted 30 days later. The oral administration of S. Typhimurium ΔSPI2 transformed with both plasmids was effective in producing antibodies against GnRXG/Q, leading to a decrease in serum testosterone levels and testicular tissue atrophy, evidenced by a reduction in the transverse tubular diameter of the seminiferous tubules and a decrease in the number of layers of the seminiferous epithelium in the testes of the inoculated mice. These results suggest that S. Typhimurium ΔSPI2 can be used as a safe and simple system to produce an oral formulation against GnRH and that Salmonella-mediated oral antigen delivery is a novel, yet effective, alternative to induce an immune response against GnRH in a murine model, warranting further research in other animal species. Full article

Objectives: Our study aimed to establish the basic reliability parameters of direct immunofluorescence test results in patients with oral lichen planus. Methods: We conducted an evaluation of individual antibody classes in the DIF and ELISA (BP180 antigen), comparing these results with the classical histopathological (HP) examination in a group of patients treated within the standard healthcare in our clinic. Results: Among 66 participants with oral changes indicative of LP, only 50% received histopathological confirmation of the LP diagnosis. Among those with a DIF profile entirely typical for LP (C3+, F+), 57.1% had a positive HP result. Fibrinogen deposits were identified in 42.4% and 36.4% of individuals with positive HP results for F1 and F2, respectively; 78.8% of patients with negative HP and 57.6% with positive HP exhibited no fibrinogen deposits. Simultaneous positivity for F1 and F2 occurred in all cases where F1 was positive. HP confirmed positive DIF for C3 in 50% of cases. Fibrinogen deposits demonstrated the highest diagnostic accuracy (61%). Sensitivity and specificity for fibrinogen deposits were 36% and 42% for F1 and 79% and 82% for F2. The positive predictive values were 67% for F1 and 67% for F2, while the negative predictive values were 58% for F1 and 56% for F2. Overall diagnostic accuracy was reported at 61% for F1 and 59% for F2. Conclusions: Our data indicate the complementarity of HP and immunological test results and the necessity of using both methods together in cases of doubt. Full article

Obesity is concurrent with immunological dysregulation, resulting in chronic low-grade inflammation and cellular dysfunction. In pancreatic islets, this loss of function has been correlated with mature β-cells dedifferentiating into a precursor-like state through constant exposure to inflammatory stressors. As mature adipocytes likewise have the capability to dedifferentiate in vitro and in vivo, we wanted to analyze this cellular change in relation to adipose tissue (AT) inflammation and adipose tissue macrophage (ATM) activity. Using our organotypic AT explant culture method combined with a double-reporter mouse model for labeling ATMs and mature adipocytes, we were able to visualize and quantify dedifferentiated fat (DFAT) cells in AT explants. Preliminary testing showed increased dedifferentiation after tamoxifen (TAM) stimulation, making TAM-dependent lineage-tracing models unsuitable for quantification of naturally occurring DFAT cells. The regulatory role of ATMs in adipocyte dedifferentiation was shown through macrophage depletion using Plexxicon 5622 or clodronate liposomes, which significantly increased DFAT cell levels. Subsequent bulk RNA sequencing of macrophage-depleted explants revealed enrichment of the tumor necrosis factor α (TNFα) signaling pathway as well as downregulation of associated genes. Direct stimulation with TNFα decreased adipocyte dedifferentiation, while application of a TNFα-neutralizing antibody did not significantly alter DFAT cell levels. Our findings suggest a regulatory role of resident ATMs in maintaining the mature adipocyte phenotype and preventing excessive adipocyte dedifferentiation. The specific regulatory pathways as well as the impact that DFAT cells might have on ATMs, and vice versa, are subject to further investigation. Full article

This study aimed to analyze the relationship between cutaneous microcirculation reactivity, retinal circulation, macrocirculation function, and specific adhesion molecules in young patients with uncomplicated type 1 diabetes. Fifty-five patients with type 1 diabetes mellitus (T1DM), aged 8 to 18 years, were divided into subgroups based on skin microcirculation reactivity. The cutaneous microcirculatory vessels were considered reactive if post-test PORH coverage increased compared to pre-test coverage. Optical coherence tomography (OCT) was conducted to detect early retinopathic changes. Macrocirculation was described using pulsatility indices (PIs) determined for common carotid (CCA) and peripheral arteries of the upper and lower limbs. The ankle–brachial index was also assessed. There were no significant differences in retinal circulation and macrocirculation between the studied subgroups. However, there were significant differences between the various subgroups concerning the age at onset of diabetes and the sP-selectin levels but not ICAM-1 and sVCAM-1. The sP-selectin differences remained true after adjusting for age at onset. The sP-selectin level was significantly higher in the subgroup of patients with non-reactive cutaneous microcirculation. The results of our study indicate that sP-selectin may be considered as an immunological marker for cutaneous abnormalities, which serve as an early indicator of endothelial dysfunction in young patients with type 1 diabetes in the absence of classical complication. Full article

Lanolin is a fatty substance derived from sheep’s fleece. The ancient Greeks used the moisturizing and skin-protective properties of this substance. The technique of industrial production of lanolin was developed in Germany in the 19th century. Since then, this natural wax has become an extremely popular base for many different cosmetic and pharmaceutical preparations intended for the treatment and care of the skin. In addition to its medicinal and cosmetic applications, lanolin is also widely used for industrial purposes. Hypersensitivity to lanolin has raised many questions and controversies for almost 100 years. Although lanolin has significant dermoprotective properties and when applied to intact skin without inflammatory changes, it lubricates it, improves its lipid barrier, and maintains proper moisture, it can also cause contact hypersensitivity when in contact with pathologically changed or damaged skin. It can, in the same person, both protect and damage the skin, depending on the condition of the skin to which the cosmetic or medicine containing lanolin is applied. The nature of the observed reactions and the circumstances of their occurrence, as well as the lack of a clear answer to the question of whether this wax causes allergies or not, make this phenomenon one of the so-called dermatological paradoxes. Although unusual reactions to lanolin have been the subject of research for many years, they still raise many questions to which there is still no clear answer. This is mainly due to the imperfection and incompleteness of the available publications. Although many different studies have been published on hypersensitivity to lanolin, most of them are retrospective analyses of the results of routinely performed epidermal patch tests or descriptions of clinical cases. Such reports and analyses, although undoubtedly very important, are a poor tool for assessing the sensitizing potential of lanolin and/or its derivatives. It is difficult to determine the causative factors, to define lanolin allergens, to investigate immunological mechanisms, or to assess the clinical significance of this phenomenon. There is a definite lack of standardized studies on the nature of lanolin hypersensitivity involving well-selected groups of patients and healthy volunteers, which would be conducted in a reproducible manner under laboratory and/or clinical conditions. As of today, lanolin hypersensitivity seems to be both an old and new problem that still remains unresolved. Full article

Background/Objectives: Considering the large number of candidates in vaccine-testing studies against different pathogens and the amount of time spent in the preclinical and clinical trials, there is a pressing need to develop an improved in vivo system to quickly screen vaccine candidates. The model of a polyester–polyurethane sponge implant provides a rapid analysis of the specific stimulus–response, allowing the study of a compartmentalized microenvironment. The sponge implant’s defined measurements were standardized as a compartment to assess the immune response triggered by the vaccinal antigen. The LBSap vaccine (composed of Leishmania braziliensis antigens associated with saponin adjuvant) was used in the sponge model to assess the antigen-specific immunological biomarker, including memory generation after initial contact with the antigen. Methods: Mice strains (Swiss, BALB/c, and C57BL/6) were previously immunized using LBSap vaccine, followed by an antigenic booster performed inside the sponge implant. The sponge implants were assessed after 72 h, and the immune response pattern was analyzed according to leukocyte immunophenotyping and cytokine production. Results: After LBSap vaccination, the innate immune response of the antigenic booster in the sponge implants demonstrated higher levels in the Ly+ neutrophils and CD11c+ dendritic cells with reduced numbers of F4/80+ macrophages. Moreover, the adaptive immune response in Swiss mice demonstrated a high CD3+CD4+ T-cell frequency, consisting of an effector memory component, in addition to a cytoxicity response (CD3+CD8+ T cells), displaying the central memory biomarker. The major cell surface biomarker in the BALB/c mice strain was related to CD3+CD4+ effector memory, while the increased CD3+CD8+ effector memory was highlighted in C57/BL6. The cytokine profile was more inflammatory in Swiss mice, with the highest levels of IL-6, TNF, IFN-g, and IL-17, while the same cytokine was observed in in C57BL/6 yet modulated by enhanced IL-10 levels. Similar to Swiss mice, BALB/c mice triggered an inflammatory environment after the antigenic booster in the sponge implant with the increased levels in the ILL-6, TNF, and IFN-g. Conclusions: The findings emphasized the impact of genetic background on the populations engaged in immune responses, suggesting that this model can be utilized to enhance and track both innate and adaptive immune responses in vaccine candidates. Consequently, these results may inform the selection of the most suitable experimental model for biomolecule testing, taking into account how the unique characteristics of each mouse strain affect the immune response dynamics. Full article

In recent years, numerous potential prognostic biomarkers for rheumatoid arthritis (RA) have been investigated. Despite these advancements, clinical practice primarily relies on autoantibody tests—for rheumatoid factor (RF) and anti-citrullinated protein antibody (anti-CCP)—alongside inflammatory markers, such as the erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). Expanding the repertoire of diagnostic and therapeutic biomarkers is critical for improving clinical outcomes in RA. Emerging evidence highlights the significance of synovial fluid biomarkers, including aggrecan, matrix metalloproteinases, glucosyl-galactosyl-pyridinoline, hyaluronic acid, S100 proteins, calprotectin, and various cytokines, as well as immunological markers. Additionally, specific components of extracellular vesicles, such as non-coding RNAs, heat shock proteins, and lipids, are gaining attention. This review focuses on molecular markers found in synovial fluid and extracellular vesicles, excluding clinical and imaging biomarkers, and explores their potential applications in the diagnosis and management of RA. Full article

The increase in honey fraud in the global market has highlighted the importance of pollen analysis in determining or confirming the botanical and geographical origins of honey. Numerous studies are currently underway to develop efficient and rapid methods for the determination of the quality, botanical, and geographical origin of honey. Typically, the physicochemical analysis of honey is used to evaluate its quality and geographical source. In this study, flow cytometry, a technique extensively employed in immunology and hematology, was first applied to analyze and characterize pollen from longan honeys from Taiwan and Thailand. The flow cytometry was employed for forward scatter (FSC), side scatter (SSC), Y610-A, and NUV450 to analyze longan honey samples from Taiwan and Thailand. Taiwan’s longan honeys were rich in pollens; however, based upon the FSC and SSC analyses, the pollens from Thai longan honeys were larger and more granular. The Y610/20 emission area was greatest in Thai pollens. The NUV450 measured in the near UV laser was also greater in Thai pollen. Additionally, honey samples were also analysed for physiochemical properties including moisture content, pH, ash content, viscosity, and hydroxymethylfurfural (HMF) for physiochemical properties of longan honey samples from both countries. The moisture content of honey from Taiwan varied between 20.90% and 23.40%, whereas honey from Thailand ranged from 19.50% to 23.50%. A total of 60% of Taiwan’s longan honey was found to have a dark amber color, and only 20% of Thai longan honey exhibited a dark amber color. Furthermore, the pH range of honey from Taiwan was found to be between 4.00 and 4.16, and the pH of Thai honey ranged from 4.01 to 4.12. The ash content of honey samples from Taiwan ranged from 0.05% to 0.23%, and Thai honey had a range of 0.01% to 0.9%. All samples were negative for the Fiehe’s test, indicating the absence of HMF. This analysis lays the groundwork for rapid identification the origins of the honey, applying flow cytometry in conjunction with physicochemical analysis to assess its quality. Full article

Background: Current challenges in meningioma treatment, including post-surgical complications and cognitive impairments, highlight the need for new treatment alternatives. Immunological interventions have shown promise. However, there is a knowledge gap in characterizing infiltrating immune cells in meningioma and their interplay. Further studies on immune cells in single-cell suspensions from digested meningioma tissues could identify targetable mechanisms for non-surgical treatment options with fewer side effects. This study aimed to optimize a protocol for faster digestion of meningioma tissues into viable single-cell suspensions and to identify infiltrating immune cell populations. Methods: We modified a commercial kit intended for whole skin dissociation to digest resected meningioma tissues into viable single-cell suspensions. Tumor-infiltrating immune cell populations were characterized using flow cytometry. Results: Flow cytometry analyses revealed that the digested tissue was composed of viable immune cells, including predominantly CD14⁺ macrophages and CD3⁺ T cells, with minor populations of CD56⁺ NK cells and CD19⁺ B cells. In both of the two patient samples tested, half of the tumor-associated macrophages were TIM-3⁺, with a small proportion co-expressing CD83. Women were more likely to have a lower proportion of immune cells, B cells, and NK cells. Female patients with a high proportion of immune cells had a higher proportion of macrophages. Conclusion: We successfully optimized a protocol for generating single-cell suspensions with viable immune cells from meningioma tissues, revealing infiltrating antigen-presenting cells with an immunosuppressive phenotype, and lymphocytes. This short protocol allows advanced analyses of tumor-infiltrating cells using techniques such as single-cell RNA sequencing and flow cytometry, which require live, dissociated cells. Full article

Periodontitis leads to immunologically mediated loss of periodontium and, if untreated, can result in tooth loss. Periodontal diseases are the most prevalent in the world and have a very strong impact on patients’ well-being and general health. Their treatment generates enormous costs. Given the above, precise, prompt, and predictive diagnosis of periodontal disease is of paramount importance for clinicians. The aim of the study was to summarize the state-of-the-art knowledge of molecular periodontal diagnostics and the utility of its clinical application. There is a great need to have diagnostic tests that not only describe the periodontal destruction that has occurred in the tissues but also allow clinicians to detect disease at a subclinical level before the changes occur. A test that would enable clinicians to follow the course of the disease and detect areas prone to exacerbation could be used to evaluate the effectiveness of ongoing periodontal therapies. Unfortunately, there is no such diagnostic method yet. A hopeful prospect is molecular diagnostics. There are numerous studies on biomarkers of periodontal disease. Point-of-care tests are also emerging. There are possibilities for processing large biological datasets (omics data). However, all of the above have a minor role in the overall single-patient diagnostics process. Despite advances in microbiological, molecular, and genetic research, the basis of periodontal diagnosis is still clinical examination enriched by the evaluation of radiological images. Full article