Total Antibody Research Articles

The Effects of Dairy Consumption on Vaccine Immune Response and Immunoglobulins: A Systematic Literature Review

Public health interest in vaccinations and immune protection has increased with the COVID-19 pandemic. Dairy products are an important source of protein and other nutrients, and there are unresolved research questions regarding the potential health impact of dairy products on the enhancement of immune response. A systematic literature review was conducted to synthesize the published literature reporting the effects of dairy interventions on: 1) the vaccine-specific immune response and 2) immunoglobulins in the absence of vaccination. To assess study validity and quality, we used the Academy of Nutrition and Dietetics Quality Criteria Checklist. Sixty-one studies (59 clinical trials, 1 cohort, 1 cross-sectional survey) were included, spanning 1983-2017. Ten trials evaluated the effect of dairy intervention on vaccine-specific IgG, IgA, IgM, vaccine-specific antibody titers, seroprotection rates, or seroconversion rates. Of these, 7 reported significant increases with dairy interventions for post-vaccine tetanus antibodies, mean change in tetanus antibody level, total antibody titers to flagellin from Salmonella Adelaide, mean antibody titers to influenza B, influenza-specific IgA and IgG levels, and seroconversion or seroprotection rates for influenza A and B. Fifty-six studies evaluated dairy’s effects on immunoglobulins without vaccinations. The results were heterogenous, with some studies reporting significant enhancement of immunoglobulins (IgA, IgE, or IgG), while others observed no differences between groups. Clinical relevance of the immunoglobulin changes was not investigated in these studies. Dairy products and their components could enhance the efficacy of vaccines. This review highlights the evidence gaps and provides a potential roadmap for additional research.

Tyrosine Sulfation at Antibody Light Chain CDR-1 Increases Binding Affinity and Neutralization Potency to Interleukine-4.

Structure and function of therapeutic antibodies can be modulated by a variety of post-translational modifications (PTM). Tyrosine (Tyr) sulfation is a type of negatively charged PTM that occurs during protein trafficking through the Golgi. In this study, we discovered that an anti-interleukin (IL)-4 human IgG1, produced by transiently transfected HEK293 cells, contained a fraction of unusual negatively charged species. Interestingly, the isolated acidic species exhibited a two-fold higher affinity to IL-4 and a nearly four-fold higher potency compared to the main species. Mass spectrometry (MS) showed the isolated acidic species possessed an +80-Dalton from the expected mass, suggesting an occurrence of Tyr sulfation. Consistent with this hypothesis, we show the ability to control the acidic species during transient expression with the addition of Tyr sulfation inhibitor sodium chlorate or, conversely, enriched the acidic species from 30% to 92% of the total antibody protein when the IL-4 IgG was co-transfected with tyrosylprotein sulfotransferase genes. Further MS and mutagenesis analysis identified a Tyr residue at the light chain complementarity-determining region-1 (CDRL-1), which was sulfated specifically. These results together have demonstrated for the first time that Tyr sulfation at CDRL-1 could modulate antibody binding affinity and potency to a human immune cytokine.

Seroprevalence of Japanese encephalitis virus-specific antibodies in Australia following novel epidemic spread: protocol for a national cross-sectional study

IntroductionJapanese encephalitis virus (JEV) is a mosquito-borne flavivirus that causes encephalitis and other morbidity in Southeast Asia. Since February 2022, geographically dispersed JEV human, animal and vector detections occurred on...

Abstract 112: Assessing the Impact of the COVID-19 Pandemic on Childhood Arterial Ischemic Stroke: An Unanticipated Natural Experiment

Introduction: The Vascular effects of Infection in Pediatric Stroke (VIPS II) study aimed to confirm prior findings that viral infection can trigger childhood arterial ischemic stroke (AIS). The COVID-19 pandemic developed midway through VIPS II enrollment, creating an opportunity to assess its impact. Methods: At 22 North American sites, we enrolled 205 children (28 days-18 years) with AIS (12/2016 - 1/2022), collected serum samples up to 7 days post-stroke, and measured clinical infection via parental interview. We defined the COVID-19 pandemic epoch as 1/2020-1/2022. We collected serum samples on 40 stroke-free controls (median [IQR] age 13.2 years [6.4, 16.5]) during the pandemic. We measured SARS-CoV2 spike IgG antibodies (present after vaccination or infection) and nucleocapsid IgG total antibodies (present after infection only; elevated for months). Results: The sites saw 117 AIS cases in 2019 (51 enrolled), 122 (44) in 2020, and 109 (52) in 2021. Comparing the pre-pandemic (N=105) and pandemic (N=100) cohorts, pandemic cases were less likely to report a recent infection (Table). Evidence of SARS-CoV2 infection was seen in 4 (10%) of controls and 26 (25%) of 100 pandemic cases (p=0.065); 10 of the 26 COVID positive cases reported a recent infection, and 7 of the 10 had a clinical diagnosis of COVID. Median (IQR) nucleocapsid IgG titers were 18.8 (12.0, 101) in the positive controls and 50.1 (26.9, 95.3) in the positive cases (p=0.33). Conclusions: The COVID19 pandemic reduced the proportion of childhood AIS cases with preceding infection, likely reflecting decreased infectious exposures while “stay at home” and masking policies were in effect. A quarter of cases in the pandemic era had positive antibodies consistent with prior SARS-CoV2 infection. Like other childhood respiratory viruses, SARS-CoV2 may act as a trigger for childhood AIS, but further investigation is needed to establish a causal role.

Long-Term Kinetics of SARS-CoV-2 Neutralizing and Anti-Receptor Binding Domain Antibodies among Laboratory-Confirmed COVID-19 Cases in Delhi National Capital Region, India: A Prospective, One-Year Follow-Up Study.

Background: This study was conducted with the objective of measuring the neutralizing and anti-receptor binding domain antibody levels against SARS-CoV-2 among laboratory-confirmed COVID-19 cases and exploring its long-term kinetics over a period of 1 year. Methods: One hundred laboratory-confirmed COVID-19 cases were recruited. Serum samples of the participants were collected within three months from the date of the positive COVID-19 report. The participants were prospectively followed up every three months for symptoms and the collection of blood samples for three additional rounds. The presence of anti-SARS-CoV-2 antibodies (IgA, IgG, and IgM antibodies), anti-receptor binding domain antibodies (anti-RBD), and neutralizing antibodies were measured. Findings: Median plaque reduction neutralization test (PRNT) titers showed a rising trend in the first three rounds of follow-up. The quantitative anti-receptor binding domain ELISA (QRBD) values showed a declining trend in the initial three rounds. However, both the PRNT titers and QRBD values showed significantly higher values for the fourth round of follow-up. Total antibody (WANTAI) levels showed an increasing trend in the initial three rounds (statistically significant). Interpretation: Neutralizing antibodies showed an increasing trend. The anti-receptor binding domain antibodies showed a decreasing trend. Neutralizing antibodies and anti-RBD antibodies persisted in the majority.

First-in-Human Evaluation of Site-Specifically Labeled 89Zr-Pertuzumab in Patients with HER2-Positive Breast Cancer.

Radioimmunoconjugates targeting human epidermal growth factor receptor 2 (HER2) have shown potential to noninvasively visualize HER2-positive tumors. However, the stochastic approach that has been traditionally used to radiolabel these antibodies yields poorly defined and heterogeneous products with suboptimal invivo performance. Here, we describe a first-in-human PET study on patients with HER2-positive breast cancer evaluating the safety, biodistribution, and dosimetry of 89Zr-site-specific (ss)-pertuzumab PET, a site-specifically labeled radioimmunoconjugate designed to circumvent the limitations of random stochastic lysine labeling. Methods: Six patients with HER2-positive metastatic breast cancer were enrolled in a prospective clinical trial. Pertuzumab was site-specifically modified with desferrioxamine (DFO) via a novel chemoenzymatic strategy and subsequently labeled with 89Zr. Patients were administered 74 MBq of 89Zr-ss-pertuzumab in 20 mg of total antibody intravenously and underwent PET/CT at 1 d, 3-4 d, and 5-8 d after injection. PET imaging, whole-body probe counts, and blood draws were performed to assess the pharmacokinetics, biodistribution, and dosimetry. Results: 89Zr-ss-pertuzumab PET/CT was used to assess HER2 status and heterogeneity to guide biopsy and decide the next line of treatment at progression. The radioimmunoconjugate was able to detect known sites of malignancy, suggesting that these tumor lesions were HER2-positive. The optimal imaging time point was 5-8 d after administration, and no toxicities were observed. Dosimetry estimates from OLINDA showed that the organs receiving the highest doses (mean ± SD) were kidney (1.8 ± 0.5 mGy/MBq), liver (1.7 ± 0.3 mGy/MBq), and heart wall (1.2 ± 0.1 mGy/MBq). The average effective dose for 89Zr-ss-pertuzumab was 0.54 ± 0.03 mSv/MBq, which was comparable to both stochastically lysine-labeled 89Zr-DFO-pertuzumab and 89Zr-DFO-trastuzumab. One patient underwent PET/CT with both 89Zr-ss-pertuzumab and 89Zr-DFO-pertuzumab 1 mo apart, with 89Zr-ss-pertuzumab demonstrating improved lesion detection and higher tracer avidity. Conclusion: This study demonstrated the safety, dosimetry, and potential clinical applications of 89Zr-ss-pertuzumab PET/CT. 89Zr-ss-pertuzumab may detect more lesions than 89Zr-DFO-pertuzumab. Potential clinical applications include real-time evaluation of HER2 status to guide biopsy and assist in treatment decisions.

Comparative Analysis of AB vs. ABO-specific Plasma for Desensitization in Blood Group O Recipients: An In Vitro Study.

Neutralizing capacity measurement (NCM) of soluble ABH substances (SAS) in plasma was assessed to guide the selection of the appropriate ABO group of fresh-frozen plasma (FFP) for plasma exchange (PE) in blood group O recipients with ABO-incompatible transplantations. Neutralizing capacity was assessed by measuring anti-A and/or anti-B titers in samples comprising one unit of O FFP and 10 O EDTA plasma samples and subtracting the binary logarithm of the titer in each group with a saline dilution. Ten EDTA plasma samples with Lewis b (Leb) antigen positivity and 10 sets of pooled FFP from each blood group were used as diluents. In O FFP, the NCM values (mean±SD) were 3.4±0.52 (2.6±0.52) and 2.6±0.52 (1.5±0.3) in B and AB for IgM (total antibody) anti-B (both P<0.001), and in the 10 O EDTA plasma samples, they were 3.9±0.88 (3.1±0.88) and 3.2±0.79 (2.4±0.97) for IgM (P=0.0013) and total anti-B (P=0.025), respectively. In vitro analysis revealed that B FFP is more effective than AB FFP in reducing IgM and total anti-B antibody titers in O recipients, regardless of Leb antigen positivity.

Development and validation of pharmacokinetics assays for a novel HER2-targeting antibody-drug conjugate (SHR-A1201): Application to its dose-escalation pharmacokinetic study

Approximately 25% of breast cancer patients with HER2 overexpression tend to have a high risk of disease progression and death. Various HER2-targeting therapies have been approved for treatment. Recently, a novel antibody-drug conjugate, SHR-A1201, is being researched and developed. For the pharmacokinetic study of SHR-A1201, suitable bioanalytical methods are needed for quantifying unconjugated cytotoxin, cytotoxin-conjugated antibodies and total antibodies. In this research, bioanalytical methods involving a highly sensitive LC-MS/MS assay for unconjugated cytotoxic payload DM1 in human plasma, ELISA strategies for DM1-conjugated trastuzumab and total trastuzumab in human serum were developed, validated and successfully applied to a phase I dose-escalation pharmacokinetic study of SHR-A1201. The pharmacokinetic properties and exposure-to-dose proportionality was evaluated for SHR-A1201. According to the bioanalytical method validation guidance, the bioanalytical methods were fully validated and the validation results met the acceptance criteria. The nonspecific binding of DM1 and dimer was avoided for the LC-MS/MS assay. In the dose-escalation pharmacokinetic study of SHR-A1201, a potential dose-proportional pharmacokinetics was observed over the dose from 1.2 mg/kg to 4.8 mg/kg. The validated bioanalytical strategies are robust and reproducible and these bioanalytical methods will contribute to better understanding of the pharmacokinetic properties of SHR-A1201.

Practical Considerations for Delandistrogene Moxeparvovec Gene Therapy in Patients With Duchenne Muscular Dystrophy

BackgroundDelandistrogene moxeparvovec is a gene transfer therapy approved in the United States, United Arab Emirates, and Qatar for the treatment of ambulatory patients aged four through five years with a confirmed Duchenne muscular dystrophy (DMD)-causing mutation in the DMD gene. This therapy was developed to address the underlying cause of DMD through targeted skeletal, respiratory, and cardiac muscle expression of delandistrogene moxeparvovec micro-dystrophin, an engineered, functional dystrophin protein. MethodsDrawing on clinical trial experience from Study 101 (NCT03375164), Study 102 (NCT03769116), and ENDEAVOR (Study 103; NCT04626674), we outline practical considerations for delandistrogene moxeparvovec treatment. ResultsBefore infusion, the following are recommended: (1) screen for anti-adeno-associated virus rhesus isolate serotype 74 total binding antibody titers <1:400; (2) assess liver function, platelet count, and troponin-I; (3) ensure patients are up to date with vaccinations and avoid vaccine coadministration with infusion; (4) administer additional corticosteroids starting one day preinfusion (for patients already on corticosteroids); and (5) postpone dosing patients with any infection or acute liver disease until event resolution. Postinfusion, the following are recommended: (1) monitor liver function weekly (three months postinfusion) and, if indicated, continue until results are unremarkable; (2) monitor troponin-I levels weekly (first month postinfusion, continuing if indicated); (3) obtain platelet counts weekly (two weeks postinfusion), continuing if indicated; and (4) maintain the corticosteroid regimen for at least 60 days postinfusion, unless earlier tapering is indicated. ConclusionsAlthough the clinical safety profile of delandistrogene moxeparvovec has been consistent, monitorable, and manageable, these practical considerations may mitigate potential risks in a real-world clinical practice setting.

Redirecting antibody responses from egg-adapted epitopes following repeat vaccination with recombinant or cell culture-based versus egg-based influenza vaccines

Repeat vaccination with egg-based influenza vaccines could preferentially boost antibodies targeting the egg-adapted epitopes and reduce immunogenicity to circulating viruses. In this randomized trial (Clinicaltrials.gov: NCT03722589), sera pre- and post-vaccination with quadrivalent inactivated egg-based (IIV4), cell culture-based (ccIIV4), and recombinant (RIV4) influenza vaccines were collected from healthcare personnel (18-64 years) in 2018−19 (N = 723) and 2019−20 (N = 684) influenza seasons. We performed an exploratory analysis. Vaccine egg-adapted changes had the most impact on A(H3N2) immunogenicity. In year 1, RIV4 induced higher neutralizing and total HA head binding antibodies to cell- A(H3N2) virus than ccIIV4 and IIV4. In year 2, among the 7 repeat vaccination arms (IIV4-IIV4, IIV4-ccIIV4, IIV4-RIV4, RIV4-ccIIV4, RIV4-RIV4, ccIIV4-ccIIV4 and ccIIV4-RIV4), repeat vaccination with either RIV4 or ccIIV4 further improved antibody responses to circulating viruses with decreased neutralizing antibody egg/cell ratio. RIV4 also had higher post-vaccination A(H1N1)pdm09 and A(H3N2) HA stalk antibodies in year 1, but there was no significant difference in HA stalk antibody fold rise among vaccine groups in either year 1 or year 2. Multiple seasons of non-egg-based vaccination may be needed to redirect antibody responses from immune memory to egg-adapted epitopes and re-focus the immune responses towards epitopes on the circulating viruses to improve vaccine effectiveness.

SARS-CoV-2 Antibody Prevalence in Unvaccinated Children and Change in Antibody Levels Between 2020-2022

After the declaration of the COVID-19 pandemic by the World Health Organization (WHO), various restrictions were implemented, and face-to-face education was suspended in schools. In some countries, COVID-19 vaccines emergency use authorization has been granted for the pediatric age group. This study aimed to investigate COVID-19 antibody level in children who have not been vaccinated against COVID-19. A total of 1,000 pediatric patients aged 0-12 years, unvaccinated against COVID-19, admitted to Necmettin Erbakan University Medical Faculty Hospital Pediatrics clinics with different symptoms between October 2020 and April 2022 were included in this study. SARS-CoV-2 S total antibody levels were tested by ELISA method from the sera samples of these children. Of the children included in the study, 535 (53.5%) were male and 465 (46.5%) were female. Anti-SARS-CoV-2 S total Ig positivity rates were 75.9% in girls and 78.5% in boys and the difference between them was not statistically significant (p&gt;0.05). When the anti-SARS-CoV-2 S total Ig positivity rates of children were evaluated according to years, 23 (43.4%) in 2020, 183 (60%) in 2021, and 567 (88.3%) in 2022 was found positive. The difference between antibody results by years was found to be statistically significant (p&lt;0.001). Median value of SARS-CoV-2 antibody levels of patients with positive antibody test in 2020 46 AU/ml (min 5, max: 214 AU/ml), in 2021 130 AU/ml (min:1, max: 250 AU/ml), and in 2022 it was found to be 250 AU/ml (min:1, max: 250 AU/ml). When the antibody levels of patients with anti-SARS-CoV-2 S Ig total positive were evaluated, the difference between years was found to be statistically significant (p&lt;0.05). In our study, it was determined that the rate of SARS-CoV-2 S total antibody positivity reached 90% in unvaccinated children aged 12 and younger. Determining the SARS-CoV-2 antibody status of children aged 12 and under who unvaccinated against COVID-19 in our country will both give information about immunization by natural immunity and will be effective in deciding and planning the need for COVID-19 vaccination for children.

Exploring the role of autoantibodies in Iraqi females with polycystic ovary syndrome

Background: Polycystic ovary syndrome (PCOS) is a prevalent endocrinopathy that affects 5-10% of women during their reproductive years. Clinicians characterize it by the occurrence of ovarian cysts, oligomenorrhea, amenorrhea, and either clinical or biochemical evidence of hyperandrogenism. Recent research suggests a potential link between autoantibodies and PCOS development. Aim of study: This study investigated the relationship between autoantibodies and Polycystic ovary syndrome (PCOS). Materials and methods: The study involved a total of 150 female participants diagnosed with PCOS, aged between 20 to 35, and a control group of 150 individuals. We utilized chemiluminescent immunoassays to measure Anti-thyroid antibodies thyroglobulin (TG) and thyroid peroxidase (TPO). We conducted IgM and IgG for anti-cardiolipin (ACL) and β-2 glycoprotein I (B2GPI) using the enzyme-linked immunosorbent assay (ELISA). To measure lupus (La), we employed coagulation time. To assess the serum levels of total antibodies Ig (M, G, A), Serum amyloid A (SAA), High-Sensitivity C-reactive protein (hs-CRP), and Procalcitonin (PCT), we used nephelometry methods. Results: In our findings, patients showed a significantly higher level of TPO autoantibodies (91.9±14.7) compared to the control group (12.4±5.1). This difference was highly significant (&lt;0.001). Similarly, we observed high levels of anti-TG antibodies in patients (50.4±10.3) compared to the control group (30.6±7.2). This points out the important role of both TPO and TG antibodies in this context. Moreover, the level of IgM for ACL was significantly higher in patients (19.2±4.2 AU/dl) than in controls (3.3±1.0 AU/dl). The level of IgG for ACL was also higher in patients (22.5±2.3 AU/dl) compared to controls (3.3±1.0 AU/dl). These increases in ACL antibodies in patients suggested a risk factor for the disease. Additionally, Serum amyloid A (SAA) and high-sensitivity C-reactive protein (hs-CRP) were significantly higher in patients, with measurements of 66±10.4 (AU/ml) and 55±12.5 (AU/ml) respectively, compared to controls who had measurements of 3.6±1.9 (AU/ml) and 2.3±0.98 (AU/ml) respectively, which was highly significant (&lt;0.001). Procalcitonin (PCT) was also highly significant in patients (40±6.3 AU/ml) compared to the control group (6.4±2.6 AU/ml). Lastly, the concentration of total IgG antibodies was significantly higher in patients (25.3±2.6 AU/dl) than in controls (4.3±1.0 AU/dl), with a significant difference (P=0.006). Conclusion: High concentrations of autoantibodies in patients suggest a risk factor and positive associations with the disease.

A Prospective Study of Seroconversion Post Covishield Vaccination in COVID-19 Warriors

Introduction: During the Coronavirus Disease 2019 (COVID19) pandemic in India, two vaccinees were predominantly administered to prevent the spread of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The first vaccine introduced in India was ChAdOx1-nCOV (Covishield), followed by BBV-152 (Covaxin). In the first phase, Healthcare Workers (HCW) were prioritised for vaccination, given their crucial role in the healthcare system. Aim: To assess the antibody response post Covishield vaccination at specific time intervals in HCWs and to determine the correlation of antibody response with age, gender, comorbidities and blood group. Materials and Methods: This prospective study was conducted at Dr. Balasaheb Vikhe Patil Rural Medical College, Pravara Institute of Medical Sciences, Loni, Maharashtra, India, over a duration of six months from January 2021 to June 2021. A total of 110 vaccinated HCWs who volunteered were included in this study. SARS-CoV-2 antibodies at specific time intervals were assessed using Ortho Clinical Diagnostic’s VITROS 3600 based on the principle of Chemiluminescent Immunosorbent Assay (CLIA). Assessment of anti-SARS-CoV-2 total and IgG antibodies was performed at 15 days, one month, one and a half months, two months, three months, and four months postfirst dose of vaccination, or in other words, 15 days and one month after the 1st dose, and 15 days, one month, two months, and three months post second dose of vaccination. For analysis, Pearson’s correlation and a regression model were performed using GraphPad Prism 8.0.2 version. Results: Fifteen days post second dose, 110 HCWs (100%) and 109 (99.09%) HCWs turned seropositive for total antibodies and IgG antibodies, respectively. It was observed that the majority of participants (33, 30.27%) with peak IgG levels in the medium range were from the age group of 31-40 years. Overall, there was a negative correlation between age and IgG antibody levels for peak IgG values (r-value=-0.224, p-value=0.019). The peak values were achieved in the majority of participants 15 days post second dose (53.6%). The difference in antibody levels based on gender was not significant (Chi-square value=3.387, p-value=0.184). No significant difference in SARS-CoV-2 IgG levels was observed between participants with co-morbidities and those without co-morbidities. Participants who developed SARS-CoV-2 infection during the study period exhibited robust antibody responses after vaccination. Conclusion: These findings help elucidate Covishield vaccinespecific antibody responses in vaccinees of different age groups, genders, blood groups, and with co-morbid conditions. The vaccine has substantially reduced the burden of disease by preventing serious illness in vaccinated HCWs during the second wave of the COVID-19 pandemic.

A novel lateral flow immunochromatographic assay using a recombinant VP2 antigen for total antibody detection of canine parvovirus-2.

Canine parvovirus-2 (CPV-2) is a viral disease of dogs causing acute hemorrhagic gastroenteritis and myocarditis with high morbidity and mortality rates. The infection is still widespread all over the world. Vaccines developed against infection have great importance in preventing infection. However, it is difficult to recommend a practical vaccination program without knowing the antibody level of a puppy. Despite widespread vaccination, difficulties in detecting the maternal antibodies in puppies remain the main cause of vaccination failure. The hemagglutination inhibition (HAI) test is the gold standard to determine the immune status of dogs for canine parvovirus 2, but the HAI test has several disadvantages such as the need for fresh porcine blood, well-equipped laboratory, and long incubation periods. In this study, for the first time we developed a colloidal gold-based competitive lateral flow assay (cLFA) system for the rapid detection of total antibodies in canine serum using CPV-2b-VP2 derived from field isolates. The recombinantly expressed capsid protein of CPV-2 in the prokaryotic expression system was used as a labeled molecule in cLFA. We carried out studies on our cLFA system using the standard antibody solution and the clinical samples from vaccinated puppy serum. We compared the results of the LFAs with the HAI test. Competitive lateral flow assay results showed good correlation with the gold standard method, the HAI test. In the developed platform, the limit of detection of the standard antibody was determined to be 375 ng mL-1, while the cut-off level of antibodies was observed to be 1 : 40 HAI titer in clinical samples. Our reported system will be a strong alternative for CPV-2 antibody-based detection applications.

Analysis of Specific IgE Distribution Characteristics of Dust Mite Allergen Components in Patients With Atopic Dermatitis

Objective: To investigate the distribution characteristics of specific immunoglobulin E (IgE) to dust mite components in patients with atopic dermatitis (AD), aiming to provide a reference basis for the development of microarray protein chips for allergen component discrimination diagnosis and personalized desensitization therapy.Methods: A total of 55 patients who visited Huishan District People’s Hospital of Wuxi City, Xishan People’s Hospital of Wuxi City, and People’s Hospital of Wuxi City from January 2023 to December 2023 were selected. The results of serum total IgE (total IgE) and specific IgE detection were analyzed.Results: Among the 55 patients, 54 (98.2%) were positive (cutoff value &gt; 0.35 Ua/mL, the normal value for serum total IgE is under 150–400 IU/mL) for serum total IgE antibodies, 52 (94.6%) were positive for Dermatophagoides pteronyssinus (Dp)–specific IgE antibodies, and 50 (90.9%) were positive for Dermatophagoides farinae (Df)–specific IgE antibodies. The protein chip results were consistent with the existing Phadia detection results.Conclusion: The development of chips based on CRD (component‐resolved diagnosis) cannot only rely on IgE binding rate to assist in precise treatment of patients but also choose personalized desensitization therapy, providing more practical recommendations for preventing cross‐allergy, avoiding allergic environments, or ingesting sensitizing foods for susceptible individuals.

Assessment of tolerance of a new product (gluten-free grain snack) in children with food allergy to gluten

The development of a technological scheme for the production of cereal snacks based on the extrusion process with preliminary enzymatic treatment of wheat flour and the use of hydrolysate as a partial replacement for gluten-free raw materials is an urgent task. The use of gluten-free cereal snacks in the diet opens up new possibilities for dietary therapy in patients suffering from various manifestations of food allergy to gluten, making their diet more diverse and meeting their individual needs. The purpose of the research was to clinically evaluate the tolerance of gluten-free cereal snacks in children with PA to gluten. Material and methods. The study included 21 patients (male and female) aged 3 to 17 years with a diagnosed food allergy, including gluten. All children received gluten-free cereal snacks according to their age: children from 3 to 7 years old (n=11) received 50 g (176 kcal) daily, and children aged 8 to 17 years old (n=10) received 100 g (352 kcal) daily for 14 days, in addition to a non-specific hypoallergenic diet excluding highly allergenic foods. On the first day of the study, the patients or one of their parents evaluated the organoleptic properties of the presented product (taste, color, smell) using a 5-point scale (1 - very bad, 5 - excellent), according to a specially designed questionnaire. The assessment of clinical symptoms over time related to skin, gastrointestinal tract, and nervous system was conducted using a second questionnaire, where parents kept a daily «observation log» reflecting the child's well-being and condition, as well as adverse events associated with taking the product. Clinical safety parameters were assessed based on the dynamics of complete blood count and blood immunological parameters (total IgE, specific IgE, and IgG4 antibodies to the studied allergens) at the beginning and end of the consumption period. Results. Daily examinations and patient questionnaires revealed no adverse reactions to the product. No significant differences in body weight were observed at the end of the study (p>0.05). In the results of clinical and immunological blood analysis before and after consuming gluten-free cereal snacks, no significant deviations were observed in the form of an increase in eosinophil count, total IgE, allergen-specific IgE and IgG4 antibodies in peripheral blood. This suggests a favorable clinical safety profile of the proposed product. Conclusion. The study found that consumption of the new product (gluten-free cereal snacks) was well-tolerated by children aged 3 to 17 years with various manifestations of food allergy: no adverse effects of the product or exacerbations of the underlying disease were observed during the 14-day period of gluten-free cereal snack consumption. Therefore, the studied gluten-free cereal snacks can be recommended for children aged 3 to 17 years with various manifestations of gluten FA, in combination with a gluten free diet.

Ameliorative effects of Lactobacillus against Aflatoxin B1.

Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P<0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p<0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.

Fasciola hepatica infection modifies IgG1 specific immune response to foot-and-mouth disease virus induced by vaccination

Fasciola hepatica, a worldwide distributed helminth, has a robust immunoregulatory effect in the host, increasing the susceptibility to secondary infections. Foot and mouth disease (FMD) is a highly contagious acute vesicular viral disease effectively controlled by vaccination in endemic regions. Despite the evidence of immunoregulatory effects, the impact of fasciolosis on the immune response induced by FMD vaccination in cattle has never been assessed. Our objective was to evaluate whether the infection by F. hepatica in cattle influences the long-term immunity elicited by the currently used commercial FMD-inactivated vaccines. Aberdeen Angus steers negative for F. hepatica were vaccinated twice against FMD virus (FMDV) during the first 6 months of age using a commercial oil vaccine formulated with A24/Cruzeiro and O1/Campos strains. When maternal antibodies against F. hepatica were weaned (18––20 months of age) animals were divided into groups of 12 and infected or mock-infected with 500 metacercariae/animal. Individual serum samples were collected at 0-, 28-, 59-, 87- and 157-days post-infection (dpi). Indirect ELISAs were used to detect A24/Cruzeiro specific bovine IgG and IgG subtypes. The total IgG antibody levels and avidity against FMDV did not show significant differences between all the groups. The commercial vaccine induced higher IgG2 than IgG1 titers in vaccinated animals. Anti-FMDV IgG1 levels significantly decreased in the infected group at 28 dpi. In addition, the avidity of IgG1 FMDV-specific antibodies at day 28 in the infected group was reduced compared to the control. These results show that F. hepatica infection modified anamnestic responses against FMDV, reducing serum IgG1 titers and avidity. To our knowledge, this is the first report of immune-regulation of F. hepatica altering the immune response of FMD vaccines, one of the most globally used animal vaccines.

Role of Xenosialylation in Post-Infectious and Post-Vaccination Complications, Including Covid-19 and Anti-SARS-CoV-2 Vaccination.

The host glycosylation mechanism, with sialic acids as a key component, is essential for synthesizing carbohydrate components in viral glycoproteins. We hypothesize a correlation between the presence of the Neu5Gc on the host tissue and the development of infectious complications, adverse vaccine reactions, and autoimmune diseases. In certain mammals, including humans, the loss of the Cytidine Monophospho-N-Acetylneuraminic Acid Hydroxylase gene (negative-CMAH) prevents the synthesis of Neu5Gc, which acts as a Mammalian-associated Carbohydrate Antigen (MCA), (XeSiAs-Neu5Gc). When negative-CMAH species consume products from positive-CMAH mammals or are exposed to non-human cell-derived medicines, Neu5Gc can be integrated into their glycocalyx through a process called xenosialylation, eliciting an inflammatory response (xenosialitis) and prompting the production of circulating anti-Neu5Gc antibodies aimed at eliminating Neu5Gc. We hypothesize that in the case of xenosialylation, neutralizing antiviral antibodies from infections or vaccinations-including those for SARS-CoV-2-may cross-react with the XeSiAs-Neu5Gc glycans, as these resemble viral envelope antigens produced by the host's glycosylation. Additionally, circulating anti-Neu5Gc antibodies may also react with other circulating antibodies, including newly formed antiviral ones with a XeSiAs-Neu5Gc-contaminated Fc region. This can lead to the serum removal of the anti-inflammatory antibodies, leaving only hyperinflammatory IgG agalactosylated antibodies. Such conditions are also seen in various inflammatory and autoimmune diseases. We hypothesize that the combination of antibody cross-reaction and the removal of the XeSiAs-Neu5Gc-contaminated Fc region anti-inflammatory antibodies may intensify severe inflammatory responses like cytokine storms and coagulopathies in COVID-19 patients and those vaccinated. Assessing serum levels of total XeSiAs-Neu5Gc antibodies could be a valuable method for identifying patients at risk of severe complications from viral infections and vaccinations, including SARS-CoV-2. This strategy may also deepen our understanding of the pathogenesis of autoimmune diseases linked to post-infectious and post-vaccination complications, particularly for viruses utilizing the host glycosylation machinery, such as SARS-CoV-2, IAV, EBV, and others.

Tlr7 drives sex- and tissue-dependent effects in Sjögren's disease.

Primary Sjögren's disease (pSD) is a systemic autoimmune disease that has the strongest female predilection of all autoimmune diseases. The underlying mechanisms that govern this sexual dimorphism, however, remain poorly understood. We hypothesized that pSD females would exhibit more robust disease as compared to males, and that Tlr7 controls distinct disease manifestations in males and females. Using a well-established pSD mouse model, we harvested exocrine glands, and pulmonary and renal tissue from males and females and quantified the inflammation present. We then collected salivary glands, spleens, and cervical lymph nodes and performed flow cytometry to assess immune populations implicated in disease. We also harvested sera to examine total and autoreactive antibodies. Our data revealed that pSD mice displayed sex-biased disease, as pSD females showed decreased dacryoadenitis, but increased nephritis as compared to males. Moreover, females exhibited increased proportions of germinal center B cells and CD4+ activated/memory T cells in the periphery. Additionally, salivary gland immune populations were altered in a sex-dependent manner in pSD. Females with pSD also displayed elevated total and autoreactive IgG as compared to males. Additionally, splenic B cell Tlr7 expression was increased in females. We next generated pSD mice that lacked Tlr7 systemically and found that ablation of Tlr7 was primarily protective in pSD females, while Tlr7-deficient pSD males showed heightened disease. Thus, pSD mice display sex-biased disease and these dichotomous manifestations are governed by Tlr7 activation. This study identifies Tlr7 as a druggable target for pSD, and highlights the importance of studying pSD disease mechanisms in both sexes.