Total IgE Research Articles

The prevalence of patients suffering from chronic spontaneous urticaria, in whom omalizumab cannot be stopped even after six years.

Omalizumab (OMA) was the first FDA-approved biological drug for severe chronic spontaneous urticaria (CSU), and until today is the only beneficial and truly safe one. The objectives were: To assess the prevalence of CSU patients in whom OMA cannot be stopped over time. We also asked if biomarkers (e.g., anti-TPO antibodies and total IgE) could assist in anticipating this issue. We used our prospective registry of 93 patients, which included CSU disease duration, the onset of OMA treatment, Urticaria Activity Score (UAS7) during follow-up, co-morbidities, serum IgE levels and the presence of anti-TPO antibodies. Finally, we assessed the response to OMA during a period of six years. Out of the 93 treated CSU patients, OMA was stopped in ten patients after six months being defined as failures. In another ten patients, OMA was discontinued after 2-4 years of therapy, achieving a remission. Seventy-three patients are still treated between 2 and 6 years, having different degrees of response. Of these, in thirty-eight (52%) patients, we could not stop OMA even after six years due to CSU relapses. The prevalence of lower serum IgE levels and anti-TPO antibody positivity was significantly higher in CSU patients in whom OMA could not be stopped. This is the first study where OMA-treated CSU patients were followed up to six years. In half of them, long-term therapy of six years is still required.

B-067 Human IgE Monoclonal Antibodies as Unique Tools for Allergy Diagnostics

Abstract Background Current blood-based tests for allergy diagnosis rely on human sera or plasma as the source of IgE antibody for method validation and calibration. Obtaining human samples with sufficiently high levels of IgE presents challenges for researchers and diagnostic companies. Here, we present a unique panel of allergen-specific human IgE monoclonal antibodies (mAb) that target inhalant, food and alpha-gal allergens and offer novel approaches for investigation of IgE responses and standardization of allergy diagnostic reagents and methods. Methods Human IgE mAb (n=30) targeting inhalant allergens from dust mite, cat and dog; food allergens from peanut, egg, milk, walnut and cashew, and galactose-α-1,3-galactose, were generated from B cells from symptomatic allergic patients with rhinitis, asthma, atopic dermatitis or red meat allergy using hybridoma technology. Total and allergen-specific IgE were quantified by ImmunoCAP (whole allergen ImmunoCAP were also performed). Allergen binding profiles and mAb titers were determined by two-site immunoassay (ELISA). Antibody purity was assessed using SDS-PAGE and Western Blot. Results SDS-PAGE showed single heavy and light chain bands under reducing conditions. Antibody recognition of target allergens was confirmed by Western Blot. Allergen-specific IgE mAb concentrations for inhalant allergens ranged: 9,000-37,000 kU/L for dust mite (n=3), 2,100-53,000 kU/L for cat (n=4), 5,300-21,000 for dog (n=2). Food allergens ranged: 16,300-49,400 kU/L for peanut (n=14), 15,700-81,000 for egg (n=2), 41,600 for milk (n=1), 7,100 for walnut (n=1) and 37,000 for cashew (n=1). Alpha-gal IgE monoclonals (n=2) ranged in concentration from 15,700-81,000 kU/L. The ratios of specific IgE to total IgE measured by ImmunoCAP vary by allergen and by individual human IgE mAb. Ratios for the inhalant mAb ranged from 0.04-1.05, for the food mAb the ratios ranged from 0.33-0.99. The two alpha-gal mAb showed relatively lower specific IgE levels (average ratio was 0.14; range=0.08-0.20). Conclusions Monoclonal IgE antibodies derived from patients presenting with allergic symptoms may provide a reliable replacement for human sera to investigate allergic disease. IgE mAb to inhalant and food allergens, and to the alpha-gal carbohydrate epitope, provide unique diagnostic markers and will offer improved accuracy for assessing patient allergy by standardizing diagnostic reagents and methods. Crystal structures of several of the human IgE mAb epitopes have recently been reported (Pomés et al, J Allergy Clin Immunol, 2024). The current panel of human IgE mAb can be expanded to include additional allergen targets, which may lead to the development of improved allergy therapeutics.

B-044 Development and Evaluation of an Assay for the Detection of Human Total IgE in Serum on the Fully Automated Chemiluminescence Immunoassay Platform - Autolumo® 2000 Plus Analyzer

Abstract Background The escalating prevalence of allergic diseases on a global scale, affecting approximately 20% of the world’s population, has become a significant health concern. Clinical determination of total IgE in serum and plasma is used as an aid to the diagnosis of allergic diseases. The tIgE CLIA Microparticles Kit is a chemiluminescent microparticle immunoassay for the quantitative determination of total IgE concentrations in human serum. This kit, integrated with the AutoLumo® 2000 Plus Analyzer, offers an automated, cutting-edge platform for total IgE detection. This streamlined combination provides clinicians with a reliable tool for comprehensive and efficient allergic diseases diagnosis. Methods The performance of tIgE CLIA Microparticles Kit was evaluated on AutoLumo® A2000 Plus automated chemiluminescence analyzer and compared to a well-characterized platform (Elecsys IgE II on Cobas e601, Roche). This assay is based upon the two-step sandwich method. During detection, the patient sample and the anti-IgE coated microparticles solution were added sequentially. Unbound components were washed off after incubation. The formation of a complex occurred with the subsequent addition of a HRP conjugated anti-IgE antibody solution. The catalysis of chemiluminescent substrates by this complex generated a measurable light signal recorded as RLUs (Relative Light Units). A 2-point calibrator was used to analyze the tIgE antibody concentrations in the serum sample. The limit of blank (LoB), limit of detection (LoD), limit of quantitation (LoQ), accuracy, repeatability, linearity, specificity, on-board stability and real-time stability of the assay kit were evaluated. Serum samples (n=220) that had been previously well characterized were used for method comparisons. Results The tIgE CLIA Microparticles assay demonstrated a measurement range of 5-2500 IU/mL, with LoB, LoD, and LoQ values of ≤3 IU/mL, ≤4 IU/mL, and ≤5 IU/mL, respectively. The accuracy of the assay kit was assessed by trueness and precision following CLSI EP15-A2 and EP5-A2 guidelines. The trueness was evaluated using WHO IRP 11/234 as the reference standard, the estimate %bias between the expected and measured mean value was ≤5%. The average %CV of the intra-run and inter-run precision were 5.00% and 3.24% respectively, with a linearity of R≥0.990. The specificity to tIgE is evident, with no cross-reactivity to IgA, IgG, and IgM (≤100 μg/mL), and minimal interference from rheumatoid factor (RF) and anti-nuclear antibodies (ANA) (<10%). Furthermore, based on the CLSI EP7-A2 guideline, it showed no endogenous interference with bilirubin (30 mg/dL), hemoglobin (250 mg/dL), and triglycerides (3000 mg/dL) and no exogenous interferences with 14 commonly used allergy treatment drugs except Omalizumab (360μg/mL). Notably, the assay remains stable on-board for 28 days and exhibits real-time stability for up to 12 months at 2-8ᵒC. Clinical evaluation, involving 220 samples covering the linearity range of 5.0-2500 IU/mL, reveals a strong correlation (R>0.990) and the bias was within 15% when compared to a reference kit according to the CLSI EP9-A2 guideline. Conclusions Autolumo® Analyzers, in conjunction with the tIgE CLIA Microparticles Assay, presents a fully automated chemiluminescence immunoassay platform for accurate and cost-effective screening to aid allergic diseases diagnostics with excellent specificity, precision, and on-board stability.

B-059 Development and Validation of Multiplex Quantitative Allergy Tests based on Automated Flow Fluorescence Immunoassay Analyzer

Abstract Background Serological allergy tests measuring allergen-specific IgE (sIgE) are simpler and safer than skin prick tests, and the results are not influenced by skin condition or human factors. Quantitative detection of sIgE can also aid in evaluating patient allergies and aiding subsequent treatment. Dymind has researched and developed novel quantitative allergy tests based on the automated flow fluorescence immunoassay analyzer system (MF1280), enabling quantitative detection of multiple sIgE in just one test with one sample. Here, we present the performance of tIgE and two specific IgE test kits using the Dymind MF1280 system. Methods This study evaluated the performance of total IgE (tIgE) and two allergen-specific IgE (sIgE) panels of inhalant and food allergens, inhalant allergen panel including d1 (Dermatophagoides pteronyssinus), d2 (Dermatophagoides farina), i6 (Cockroach), e1 (Cat epithelium), e5 (Dog epithelium), w1(Common ragweed), and food allergen panel including f1 (Egg white), f2 (Cow’s milk), f3 (cod), f23 (Crab), 24 (Shrimp), and f27 (beef). Evaluation criteria included precision, linearity, limit of detection (LOD) and method comparison. As clinical performance of 425 clinical samples collected from 2 hospital laboratory in China was evaluated by both of Dymind allergen IgE MF1280 system and Phadia ImmunoCAP. Results The validated linearity of total IgE and sIgE were 2.0-5000.0 IU/mL and 0.1-100.0 IU/mL respectively. The LOD of total IgE and sIgE determined to be 2 IU/mL and 0.1 IU/mL respectively. The precision was evaluated by assaying a low and a high samples of every sIgE assay and the result of d1, d2, i6, e1, e5, w1, f1, f2, f3, f23, f24, f27 were 3.73% / 1.34% (low sample / high sample), 3.26% / 1.95%, 2.18% / 0.87%, 1.31% / 1.40%, 3.63% / 1.13%, 2.84% / 4.76%, 4.50% / 1.53%, 5.97% / 1.64%, 3.46% / 1.11%, 1.58% / 1.73%, 3.77% / 1.28%, 1.78% / 0.96% respectively. The total mean coefficient of measurements was below 7% to ensure high precision in multiplex quantitative analysis. Method comparison of 425 clinical samples between Phadia ImmunoCAP and Dymind allergen IgE showed total concordance rate was over 85% except f2 (over 80%). Conclusions The Dymind Allergen-specific IgE test kits on the automated flow fluorescence immunoassay analyzer system provide fast and accurate in vitro diagnostic detection of tIgE and sIgE for aiding diagnostics specific allergens sensitization and treatment for allergy patients, with good precision and excellent agreement compared to ImmunoCAP. The MF1280 system has advantages including random access, fully automated, multiplex quantitative detection of allergen sIgE with reducing assay time, sample consumption and lower costs.

B-337 Mass Spectrometry to evaluate treatment response in patients with Multiple Myeloma. A comparative study versus Immunofixation

Abstract Background The International Myeloma Working Group (IMWG) defines treatment response categories based on monoclonal protein (MP) changes by serum protein electrophoresis (SPE) and immunofixation (sIFE). Patients that after treatment remain positive by sIFE while SPE is negative, are categorized as very good partial response (VGPR) and, if sIFE becomes negative, patients would be considered in complete remission (CR). However, the interpretation of sIFE is qualitative and subjective. In this study we evaluate the discrepancies between sIFE and Mass Spectrometry (MS) to evaluate tumor burden and the accuracy of treatment response and it´s clinical impact in terms of progression free survival (PFS). Methods Patient samples were automatically prepared on the EXENT-iP®500 liquid handler, and sample spots were analysed using the EXENT-iX®500 MALDI-TOF mass spectrometer. Light chain spectral peaks were identified by EXENT-iQ® software and then quantified indirectly in combination with total immunoglobulins concentrations by immunoturbidimetry in the Optilite® platform (The Binding Site, part of Thermo Fisher Scientific). MPs were also identified and quantified by SPE and sIFE. Results From the 430 samples from the GEM12, GEM14 and GEM-CESAR clinical trials analyzed at pre-determined time points to evaluate treatment response (post-Induction, post-ASCT, post-cons, 1st and 2nd year after maintenance), MS identified the M protein in 45% of cases (193/430) and sIFE was positive in 32% (137/430). Combining the results of both methods, we found that MS and sIFE were concordant in 76% of cases (326/430), 26.28% IFE+/ MS+ and 49.53% IFE-/ MS-. Considering sIFE as a reference, the negative predictive value (NPV) of MS was 0.8987, with a sensitivity and a specificity of 0.8248 and 0.7270, respectively (p<0.0001). Nevertheless, the positive predictive value (PPV) was 0.5855 which could be explained by the discordances observed between IFE and MS techniques (5.58% of patients IFE+/MS- and 18.6% of patients IFE-/MS+). Indeed, when the 137 patients in VGPR (IFE+ patients) were analyzed according to MS status we found that patients IFE+/ MS+ had a trend to an inferior mPFS in comparison to IFE+/MS- (5.96 years vs not reached). These discordances were even more noticeable when comparing the 293 patients in CR (IFE- patients) with MS status, since IFE-/ MS+ patients had a mPFS of 4.65 years vs mPFS not reached for IFE-/ MS- patients (p=0.0001). Conclusions Our data confirms that IFE sensitivity limitations can affect the evaluation of treatment response in patients with MM, being more noticeable when the presence of MP band in sIFE is not clear because is very fuzzy or oligoclonal bands appear. Consequently, heterogeneity in interpretation practices across labs is a major cause of misattributed response categories, especially on those patients achieving treatment responses >VGPR. In an era of highly effective MM treatments that induce quick and deep responses, we believe IMWG response criteria should be updated with an MS response category.

B-336 Analytical verification of the EXENT® solution for the identification an quantitation of monoclonal immunoglobulins by mass spectrometry

Abstract Background Recently a new mass spectrometry (MS) technology, EXENT® solution (The Binding Site, part of Thermo Fisher Scientific), for the identification and quantification of IgG, IgA and IgM intact monoclonal immunoglobulins (M-Ig) has been launched in the European markets. In this study we present the results of onsite verification study performed in our hospital. Methods Study design, using patients serum samples, including data analysis were based on the Clinical & Laboratory Standards Institute (CLSI). Relevant standards used were: Precision-CLSI EP15-A3:2014, Linearity-EP06 ED2:2020 and Comparison - EP09C ED3:2018. Samples were automatically prepared on the EXENT-iP®500 liquid handler, sample spots were analysed using the EXENT-iX®500 MALDI-TOF mass spectrometer. Light chain spectral peaks were identified by EXENT-iQ® software and then quantified indirectly in combination with total immunoglobulins concentrations by immunoturbidimetry in the Optilite® platform (The Binding Site, part of Thermo Fisher Scientific). We compared the results of MS with capillary electrophoresis (CZE) (IgG: n=82; IgA n=19; IgM n=19) and immunofixation (sIFE) (n=541). Results Precision was, for IgG with concentrations between 50.77 g/L and 3.51 g/L of 4.14% and 4.47%, for IgA between 61.88g/L and 5.02 g/L of 1.42% and 7.17%, and for IgM between 68.32 g/L and 7.30 g/L of 5.32% and 8.44%, respectively. Deviation from linearity of this assay, using serially diluted serum samples over the following ranges for IgG (65.18 g/L - 0.23 g/L), IgA (48.91 g/L - 0.26 g/L) and IgM (49.65 g/L -0.23 g/L) was <10% for all isotypes. M-Ig concentration comparison for specimens with detectable IgG, IgA or IgM M-Ig with CZE, assessed by Passing Bablok regression was IgG g/L MS = -0.9756 + 1.523x CZE g/L (slope -2.058 to -0.3322 and intercept 1.331 to 1.679), IgA g/L MS = -2.114 + 1.646x CZE g/L (slope -12.50 to -0.5059 and intercept 1.105 to 3.296), IgM g/L MS = -1.886 + 1.935x CZE g/L (slope -5.205 to -0.3571 and intercept 1.239 to 2.462). All samples MS g/L = -1.182 + 1.556x CZE g/L (n=120). Agreement between EXENT/IFE was of 99.1% for IgGK(209/211), 97.2% IgGL(105/108), 100% AK(97/97), 100% AL(51/51), 100% MK(6/6), 100% ML(3/3), 89.2% free K(33/37), 89.3% free L(25/28), 66,7% non-secretory(4/6). Discordances intact immunoglobulin was due to samples with two M-Ig, where IFE wasn’t able to identify the major clone, for free K, free L was due to EXENT’s ability to identify the heavy chain that matched with the light chain, and for non-secretory patients due to EXENT was able to identify one M-Ig in 2/6 samples. Conclusions These on-site verification results demonstrate that the EXENT solution has a wide measuring interval, to detect M proteins with very low concentrations and to provide stable and reproducible performance with a high level of accuracy for the detection and typing of monoclonal immunoglobulins. However, the bias observed against CZE to quantify M-Ig would be justified for the lowest CZE sensitivity, CZE variability and also due to the CZE quantification uses the total protein concentration to quantify M-Ig.

Role of specific immunoglobulin-E in chronic rhinosinusitis: Its clinical relevance according to nasal challenge test

BackgroundGuidelines for chronic rhinosinusitis (CRS) propose total IgE and eosinophils as important biomarkers to identify type-2 inflammation. Despite the fact that specific IgE (sIgE) have been identified as a clinical predictor in some type-2 diseases for different clinical outcomes, its role in CRS has yet to be explored in detail. ObjetiveTo describe systemic and local sIgE in CRS and explore its possible association with clinical outcomes using nasal challenge tests (NCT). MethodsIn CRS patients, we measure total IgE, serum sIgE (SsIgE) and nasosinusal sIgE (NsIgE) against 9 allergenic sources; Der p, Der f, Blo t, Can f, Fel d, Per a, grasses, Staphylococcus enterotoxin A, and B. NCT was done using the allergen with the higher sIgE prevalence (Der p). ResultsA total of 174 patients were included. Prevalence of SsIgE was 52.8% and NsIgE 46.5%; Der p was the principal allergen for SsIgE and NsIgE. The presence of nasal polyps, asthma comorbidity, NSAID hypersensitivity, and hyposmia, were significantly associated with the presence of SsIgE and NsIgE but not with total IgE. NCT-Der p was performed in 73 CRS patients, being positive in 33 (45.2%). SsIgE have the best diagnostic accuracy (79.4%) to predict NCT results (NsIgE 67.5% total IgE 52%). ConclusionSpecific IgE is a better biomarker in CRS than total IgE. Patients with clinically relevant SsIgE have a pheno-endotype associated with different clinical outcomes. Considering the clinical relevance of SsIgE demonstrated by NCT, interventions like allergen immunotherapy in CRS must be study.

Cost-effective allergy screening: a retrospective observational study.

Background. Allergies represent a substantial health concern affecting individuals across all age groups. Diagnostic screenings, such as phadiatop and phadiatop infant, are employed to identify specific IgE antibodies associated with allergic reactions. This study delves into the relationship between total IgE levels and screening test outcomes, with the objective of establishing a total IgE threshold capable of predicting the likelihood of negative results in these screenings. Methods. This retrospective observational study included adults and children under 15 years old who underwent total IgE tests in addition to phadiatop and phadiatop infant screenings from January 2018 to December 2022. Exclusion criteria were applied to patients with insufficient serum samples or those whose IgE determinations or screening tests had been invalidated according to standard laboratory protocols. Results. Data analysis uncovered a robust correlation between total IgE levels and screening test outcomes. Additionally, thresholds of 20 UI/mL and 28 UI/mL were pinpointed for total IgE levels, below which the likelihood of obtaining a positive result in phadiatop or phadiatop infant, respectively, significantly decreased. Conclusions. These findings present cost-effective strategies for healthcare practitioners by recommending the initial use of total IgE testing. Subsequently, reflex testing with phadiatop or phadiatop infant, depending on the IgE value, could be considered.

Immunological, Antioxidant, Growth Responses, and Disease Resistance of Rainbow Trout, Oncorhynchus mykiss, with Feeding Diets Supplemented with Lactobacillus salivarius and Lutein

Abstract The aim of the present study was to assess the effects of dietary supplementation with Lactobacillus salivarius (LS) ATCC 11741 and lutein (LU) on immunological, antioxidant, and growth responses, and resistance against Yersinia ruckeri infection in rainbow trout (Oncorhynchus mykiss). There were seven experimental diets containing un-supplemented diet (CTL), L. salivarius at 1 × 106 (LS6) and 1 × 108 (LS8) CFU/g, lutein at 50 (LU50) and 150 (LU150) mg/kg, L. salivarius at 1 × 106 CFU/g plus lutein at 50 g/kg (LS6+LU50), and L. salivarius at 1 × 108 CFU/g plus lutein at 150 g/kg (LS8+LU150). These diets were provided to the fish (N= 735; 14.0±0.38 g). After 60 days of feeding, all experimental treatments exhibited significantly higher growth performance, compared to the CTL treatment. The intestinal protease activity and the intestinal lactic acid bacteria population in the probiotic-treated fish were significantly higher than in the CTL fish. The intestinal lipase activity was only higher in the LS6+LU50 treatment, compared to the CTL treatment. Compared to the CTL treatment, the intestinal total bacterial count, and serum superoxide dismutase significantly increased in the LS6+LU50 and LS8+LU150 treatments. The serum catalase significantly increased in LU150, LS6+LU50, and LS8+LU150 treatments, and serum glutathione peroxidase significantly increased in the lutein-treated fish. All experimental treatments, particularly LS6+LU50 and LS8+LU150, had significantly lower serum malondialdehyde levels, compared to the CTL treatment. The levels of serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase in LU50, LS6+LU50, and LS8+LU150 treatments were significantly lower than the CTL. All experimental treatments showed significantly higher serum lysozyme (LYZ), complement, blood respiratory burst activity (RB), and skin mucus alkaline phosphatase (ALP), compared to the CTL treatment. The highest serum LYZ, C4, total immunoglobulin (Ig), blood RB activity, and skin mucus peroxidase, ALP, and Ig were observed in the LS6+LU50 and LS8+LU150 treatments. All the experimental treatments, especially LS6+LU50, exhibited significantly lower mortality after the bacterial challenge compared to the CTL. In conclusion, dietary supplementation with 1 × 106 CFU/g L. salivarius and 50 mg/kg lutein can maximally improve growth performance, digestive enzymes, antioxidant parameters, immune responses, intestinal lactic acid bacteria, and resistance against yersiniosis in rainbow trout.

Dietary Artichoke (Cynara scolymus) Extract Ameliorated the Growth Performance, Humoral Immune Parameters and Resistance Against Aeromonas hydrophila in Goldfish (Carassius auratus)

Abstract This trial investigated the efficacy of artichoke (Cynara scolymus) extract (AE) on the growth performance, immunity, antioxidant parameters, and resistance against Aeromonas hydrophila of goldfish (Carassius auratus). For this purpose, a total number of 470 goldfish with initial weight 5.70±0.2 g were fed with four experimental diets including 0 (T0), 100 (T1), 150 (T2), and 200 (T3) mg kg−1 diet AE for 8 weeks. At the end of feeding trial, growth performances, serum immune parameters, and mucus antioxidant enzymes were measured. Fish were challenged with A. hydrophila, and the antioxidant and immunity-related gene expression were investigated. Based on the results, the highest final weight (FW) and weight gain (WG) were attained in T2 and T3 (P<0.05). Immune factors including ACH50, lysozyme, and total immunoglobulin in T2 and T3 showed the highest values (P<0.05). The expression of GR, IL1 β, TNF α, HSC70, HSP70, and HSP90 β genes in T1, T2, and T3 were higher than the control (P<0.05). The GST expression was significantly enhanced in T2 (P<0.05). The present study demonstrated that the administration of AE, especially at doses of 150 mg kg−1, could improve the growth, immunity, and antioxidant parameters, as well as enhance disease resistance against A. hydrophila in goldfish.

A study on serum zonulin in chronic spontaneous urticaria patients

hronic spontaneous urticaria (CSU) is a widespread disease with a complicated heterogenous pathophysiology. Increased intestinal permeability i.e., leaky gut has been linked to the pathology of many diseases. Zonulin was recently used as a marker for leaky guts. This study aimed to assess the relation between serum zonulin level and CSU and its possible relationship with disease activity. This was a comparative cross-sectional study, which included 97 CSU adult patients and 87 apparently healthy controls. CSU patients had significant lower zonulin level than controls (p<0.001). The median of serum zonulin level was equal to 2.93 ng/ml with interquartile range (IQR) (1.40-4.19) in the CSU group and of 3.92 ng/ml with IQR (2.97-4.69) in the control group. We found a positive correlation between serum zonulin and C-reactive protein with Pearson correlation coefficient of 0.2, (p=0.04). No significant correlation was found between serum zonulin level and urticaria activity score 7 or total immunoglobulin E level. In conclusion, this study found that serum zonulin level is lower in CSU patients than in controls which could be attributed to food restriction, severity of the CSU disease and/or drug intake in the CSU cases.

Persistently Low IgG2 Levels in a Subset of Patients Following Hematopoietic Cell Transplantation.

Infectious diseases remain a major cause of morbidity and mortality after hematopoietic cell transplantation (HCT). Secondary hypogammaglobulinemia is a risk factor for infectious diseases. Total immunoglobulin G (IgG) levels and the history of infectious diseases are an integral part of determining the indication for immunoglobulin replacement therapy. The clinical significance of IgG2 levels is not well established. Guidelines recommend using pathogen-specific IgG to evaluate patients with potential secondary immunodeficiency. However, it is difficult in practice to perform such testing. IgG2 may correlate well with pathogen-specific IgG but the clinical significance of IgG2 is not well established. To assess the prevalence of low IgG2 levels with normal IgG after HCT, we cross-sectionally measured the levels of several immunoglobulins, including IgG, IgA, IgM, and IgG2, after HCT, and we assessed the correlation between them. Among 121 patients who underwent cross-sectional measurements of IgG, IgA, IgM, and IgG2 levels after HCT, 114 had normal IgG2 levels (normal IgG2 group, ≥ 100mg/dL) and 7 had low IgG2 levels (low IgG2 group, < 100mg/dL). These 7 patients were allogeneic HCT recipients. All 7 patients with low IgG2 had cGVHD and 4/7 patients had normal total IgG levels. IgG2 levels may be low even in patients with normal IgG levels years after allogeneic HCT. Therefore, our study suggests that when patients develop infectious diseases, especially multiple episodes, it is recommended to measure IgG2 levels to exclude the possibility of secondary hypogammaglobulinemia after allogeneic HCT.

EQUAL ABPA Score 2024: A Tool to Measure Guideline Adherence for Managing Allergic Bronchopulmonary Aspergillosis.

Allergic bronchopulmonary aspergillosis (ABPA) is a complex lung disease associated with significant morbidity. The ABPA Working Group (AWG) of the International Society for Human and Animal Mycology (ISHAM) revised their management guidelines in 2024, but there is currently no standardised tool to assess adherence to these recommendations. We extracted key recommendations from the updated 2024 ISHAM-AWG guidelines, focusing on critical areas: screening and diagnosis of ABPA, managing acute and treatment-dependent ABPA, and monitoring treatment response. Each item was assigned a score ranging from zero to three. We assigned negative scores to interventions not recommended by the guidelines. We identified 38 items indicative of optimal clinical care for patients with ABPA. The score for screening asthmatics for ABPA was set at three points. For diagnosing ABPA, 16 items were included, with a score ranging from 12 to 16 points, depending on the specific components used (predisposing conditions, serum A. fumigatus-specific IgE and IgG, serum total IgE, blood eosinophil count and chest computed tomography). The management of acute ABPA comprised 11 items, with a maximum score of three points. For treatment-dependent ABPA, there were nine items (scores ranging from -3 to 6). Follow-up care comprised 10 items with a maximum score of 10-13 points, covering imaging, spirometry, testing serum total IgE levels and therapeutic drug monitoring. The EQUAL ABPA score has been developed as a comprehensive tool to quantify guideline adherence. Future studies will evaluate to which extent guideline adherence is associated with improved clinical outcomes for patients with ABPA.

Unveiling Potent Anti-Asthmatic Effect of Curcumin in Combination with Salmeterol in Swiss Albino Mice

Background: Asthma is a long-term inflammatory respiratory condition marked by alterations in the airways and an increase in inflammatory cell infiltration. It has been observed that Curcumin possesses immune-modulating, anti-inflammatory, and relaxing properties for smooth muscle in the airways. Salmeterol is believed to ease the smooth muscles of the airways. Objective: Swiss Albino mice were used in the research to examine the combination anti-asthmatic effects of Curcumin and Salmeterol in asthma produced by ova albumin and milk induced eosinophilia and leucocytosis. Methods: The mice received pre-treatment with Curcumin (10 mg/kg, 20 mg/kg intraperitoneally) as well as Salmeterol (5 mg/kg) after being stimulated with an Ovalbumin (OVA) challenge and milk. After the induction period, various hematological, biochemical, molecular (ELISA), and histological analyses were performed. Results: The findings demonstrated that the combined treatment decreased the animal’s overall leukocyte and eosinophil numbers in a manner that was dose-dependent. Additionally, the therapy reduced albumin and overall protein amount in serum, BALF and lung tissues, facilitated changes in haematological parameters, and reduced the rise of Th2 cytokines (IL-4, TNF-α, IL-13) levels that is induced by OVA in lungs and BALF, total IgE level in serum. The combined action of Curcumin and Salmeterol reduced OVA-induced inflammatory influx and ultrastructural abnormalities, according to histopathological evaluation. Conclusion: The findings of this investigation demonstrate that curcumin and salmeterol together possess anti-asthmatic effects through suppressing Th2 triggered immune response and possessing an anti-inflammatory effect and anti-allergic effect. Thus combination of treatments might be a novel technique for managing asthma.

Interactive Effects of Dietary Probiotic and Succinic Acid on the Growth Performance, Digestive Enzyme Activities, Immunomodulation, Antioxidative Capacity, and Disease Resistance in Rainbow Trout (Oncorhynchus mykiss) Juveniles

Abstract The current study was carried out to explore the effects of lactofeed (LA) as a multi-strain probiotic and succinic acid (SA) on growth variables, gut lactic acid bacteria count, digestive enzymes, innate immune responses, antioxidant capacity, and resistance against yersinia ruckeri in rainbow trout juveniles (Oncorhynchus mykiss). Three hundred sixty healthy rainbow trout juveniles (13.21±0.41 g) were randomly divided into twelve tanks (300 L) as four experimental groups. They were fed with basal diet (Control; BD), FT1: BD + 1 g/kg LA, FT2: BD + 5 g/kg SA, and FT3: BD + 1 g/kg LA + 5 g/kg SA for eight weeks. According to the results, final weight (FW), weight gain (WG), protein efficiency rate (PER), and feed conversion rate (FCR) revealed a remarkable discrepancy compared to the control group. In addition, dietary inclusion of LA (FT1 and FT3) significantly increased the specific growth rate (SGR). Dietary supplementation of LA+SA (FT3) remarkably improved total bacteria count (TBC) and amylase activity compared to the unsupplemented group. Gut lactic acid bacteria (LAB) count and digestive protease activity in all supplemented fish were remarkably higher than in unsupplemented group. Blood immunological factors including white blood cell (WBC) count, total immunoglobulin content (Ig), and the activity of lysozyme (LYZ), alternative complement pathway (ACH50), and acid phosphatase (ACP) were significantly enhanced in the supplemented groups. Neutrophil (NEU) count, skin mucus Ig and hepatic glutathione peroxidase (GPX) increased in SA (FT2 and FT3) treatments. In addition, monocyte (MON) count and skin mucus LYZ activity were significantly elevated following feeding with the FT3 diet. Other immunological parameters of skin mucus including protease activity, alkaline phosphatase (ALP) activity, and ACH50 activity, as well as hepatic superoxide dismutase (SOD) and catalase (CAT) activities increased in fish fed with diets containing LA and/or SA. Malondialdehyde (MDA) value was remarkably decreased in all supplemented rainbow trout compared to the fish fed with BD. Disease resistance against Y. ruckeri in fish fed with supplemented diets significantly improved with respect to the results obtained in the control specimens. Overall, dietary LA+SA supplementation was beneficial to improve growth performance, gut LAB count, digestive enzyme activities, innate immune responses, antioxidant capacity, and disease resistance in rainbow trout. According to these findings, 1 g/kg LA + 5 g/kg SA is suggested for adding to rainbow trout diet.

Effect of betaine supplementation on immunity, energy, and lipid metabolites of growing Murrah buffalo calves

This study aimed to observe the effect of betaine (BET) supplementation on the growth, immunity, energy, and lipid metabolites of Murrah buffalo calves. Twenty-one Murrah buffalo calves were taken and randomly distributed into three groups (n=3) after blocking by body weight (98.70±1.31 kg) and age (8.12±0.55 months). The feeding regimen was the same in all the groups except that the Treatment groups were supplemented additionally with 0.0 (B0), 7.0 (B7), and 14.0 (B14) g/d BET in three respective groups for 90 d of the experiment period. Total leukocytes (TLC), neutrophils, lymphocytes, total immunoglobulin (TIG), total antioxidant activity (TAA), energy, and lipid metabolites were observed at fortnightly intervals during 90 d experimental periods. Adding BET up to 14 g/d did not exert (P&gt;0.05) any effect on total leukocytes and neutrophils concentration. Lymphocytes, TIG, and TAA were increased significantly (P&lt;0.05) in BET received groups either dose 7 or 14 g/d than in the control. No effect of dietary addition of BET was observed on the glucose, non-esterifies fatty acid (NEFA), total cholesterol (TCH), and high-density lipoprotein cholesterol (HDL-CH). The results of our study indicate that supplemental BET may play a role in regulating the immunity of growing Murrah buffalo calves.

Allergocomponent diagnostics in the management of comorbid patients

Polyvalent sensitization (simultaneous formation of antibodies to allergens from several sources, for example, to cat allergens, birch pollen, strawberry allergens), which can occur at any age, concerns a significant number of patients with allergic diseases. The diagnostic search for a causal allergen is difficult in patients with polyvalent sensitization and/or cross-reactions to allergens of unrelated groups of allergens. We have carried out work on the detection of polysensitization in patients by the ALEX2 molecular diagnostic method, which detects specific antibodies to 300 allergens (120 allergens and 180 allergenic molecules + total IgE) in one study. The presented clinical examples clearly show the possibility of identifying the spectrum of causal allergens to clarify the diagnosis, which subsequently makes it possible to prescribe the right treatment to prevent the development of complications, which is especially important for comorbid patients

Oral food challenge test results of patients with food allergy with specific IgE levels >100 UA/ml.

Specific IgE level (sIgE) is an important factor indicating sensitization status in children with food allergies (FAs). The present study aimed to clarify oral food challenge test (OFC) results in children with FAs with sIgE levels ≥100 UA/ml compared with those in children with sIgE <100. The retrospective study analyzed patients who underwent OFC with egg white, cow milk and wheat at Gifu Prefectural general medical center, Gifu, Japan between July 2017 and March 2023. Clinical history, total IgE (tIgE), sIgE and correlation between sIgE, sIgE/tIgE and eliciting dose as the amount of intake protein were examined. In the <100 group, positive OFC showed significantly higher sIgE for egg white, ovomucoid and casein than negative OFC (P<0.05); however, there was no significant difference between positive and negative OFC in the ≥100 group. In the <100 group, positive OFC showed significantly higher sIgE/tIgE for ovomucoid, milk and casein than negative OFC (P<0.05); however, there was no significant difference in sIgE/tIgE between positive and negative OFC in the ≥100 group. There was a significant negative correlation between eliciting dose and sIgE for egg white and wheat (P<0.05). For milk and wheat, there was no significant difference between ≥100 group and the <100 group with regard to positive rates in the OFC. Therefore, OFC may be safely performed by decreasing total challenge dose for the ≥100 group.

Correlation between forkhead box P3 (rs3761548) gene polymorphism and serum interleukin13 as biomarkers of severity in Egyptian allergic conjunctivitis: a retrospective study.

The genetic variants that alter human Forkhead Box P3 (FOXP3) function may have a part in the establishment of allergic conjunctivitis. Our study aimed to evaluate the FOXP3 polymorphism, serum interleukin13 (IL13) and total immunoglobulin E (IgE) levels in allergic conjunctivitis and assess their role as biomarkers for allergic conjunctivitis risk and severity. This study included 52 cases and 52 controls. Blood samples were taken from allergic conjunctivitis patients and controls for total IgE, IL13 measurement and detection of FOXP3 (rs3761548) gene polymorphism. There was a statistically significant difference between the allergic conjunctivitis group and healthy control group regarding FOXP3 (rs3761548) polymorphism with those have AA genotype are 12 times at risk for allergic conjunctivitis and A allele increases the risk of allergic conjunctivitis by about 4 times. There was statistically significant difference between mild/moderate and severe allergic conjunctivitis regarding FOXP3 (rs3761548) polymorphism with those have AA genotype are 53 times at risk for severe allergic conjunctivitis and A allele increases the risk of severe allergic conjunctivitis by about 6 times. Also, there was a significantly higher value of total IgE IU/ml, IL13 Pg/ml value in severe allergic conjunctivitis compared to moderate/mild allergic conjunctivitis. The best cutoff values of total IgE and serum IL13 for detecting the severity of allergic conjunctivitis were ≥320 IU/ml and ≥40 Pg/ml and the area under the curve were 0.89 and 0.95 respectively. The research significantly contributes to find correlation of FOXP3 polymorphism, total IgE and IL13 with risk and severity of allergic conjunctivitis which are limited in the literature on the perceived value relevance of FOXP3 polymorphism in allergic conjunctivitis risk and severity.

Correlation between Serum Vitamin D Levels and Allergic Rhinitis Severity at Vajira Hospital: A Cross-Sectional Study

OBJECTIVE: Vitamin D affects the immune system and inflammatory process, and its deficiency is significantly associated with an increased prevalence of allergic rhinitis (AR). Therefore, the correlation between the serum 25-hydroxyvitamin D levels and the severity of AR symptoms must be investigated.METHODS: This research utilized a cross-sectional approach, focusing on patients aged 18 years and above diagnosed with AR at the Vajira Hospital, Navamindradhiraj University, Bangkok, Thailand from August 15, 2023, to January 15, 2024. The relationship between AR severity and serum vitamin D levels was analyzed using Pearson correlation coefficient. AR severity on symptoms and quality of life was assessed using the Thai version of Sinonasal Outcome Test-22 (SNOT-22) and Rhinoconjunctivitis Quality of Life questionnaire-36 (RCQ-36) questionnaires. Serum-specific immunoglobulin E (IgE) and total IgE levels were also measured to examine their relationship with the serum vitamin D level.RESULTS: Among the 58 participants, 34 (58.60%) had vitamin D deficiency. These individuals with vitamin D deficiency exhibited significantly higher AR severity compared with those without vitamin D deficiency. The mean difference between these groups was 21.90 points for SNOT-22 scores (95%CI: 17.71–26.69, p-value &lt; 0.001) and 19.17 points for RCQ-36 scores (95%CI: 12.08–27.34, p-value &lt; 0.001). Further analysis revealed a significant inverse correlation between serum vitamin D level and AR severity, with Pearson correlation coefficients of −0.72 for RCQ-36 (95%CI: −0.82 to −0.56, p-value &lt; 0.001) and 0.80 for SNOT-22 (95%CI: −0.88 to −0.68, p-value &lt; 0.001). No correlation with serum vitamin D level was found for serum-specific IgE and total IgE levels.CONCLUSION: A significant inverse relationship existed between serum vitamin D levels and AR severity. No correlation with serum vitamin D level was found for serum-specific IgE and total IgE levels.