High Antibody Levels Research Articles

Immunogenicity Assessment of a 14-Valent Human Papillomavirus Vaccine Candidate in Mice

Background: Cervical cancer ranks as the fourth most common cancer affecting women globally, with HPV as the primary etiology agent. Prophylactic HPV vaccines have substantially reduced the incidence of cervical cancer. Methods: This study assessed the immunogenicity of SCT1000, a 14-valent recombinant virus-like particle (VLP) vaccine developed by Sinocelltech, Ltd. using pseudovirion-based neutralization assays (PBNAs) and total IgG Luminex immunoassays (LIAs). Currently in phase III clinical trials in China, SCT1000 targets the same HPV types as Gardasil 9®, plus five additional high-risk types, thereby covering twelve high-risk HPV types implicated in 96.4% of cervical cancer cases. Results: In murine models, a dose of 1.85 μg per mouse was identified as optimal for evaluating SCT1000’s immunogenicity in a three-dose regimen, as measured by PBNA and total IgG LIA across all 14 HPV types. SCT1000 induced high levels of protective antibodies, which were sustained for at least four months following the third dose. The vaccine also demonstrated stable and consistent immunogenicity in mouse potency assays under both long-term and accelerated conditions. Additionally, our studies revealed a strong correlation between the two serological tests used. Conclusions: SCT1000 elicited robust, durable, and consistent humoral immune responses across all 14 HPV types, indicating its potential as a broad-spectrum vaccine candidate against HPV types 6/11/16/18/31/33/35/39/45/51/52/56/58/59. The significant correlations observed between PBNA and total IgG LIA support the use of the Luminex-based total IgG method as a reliable and effective alternative for immunogenicity assessment in preclinical and future clinical vaccine development.

Modified PEG-Lipids Enhance the Nasal Mucosal Immune Capacity of Lipid Nanoparticle mRNA Vaccines

Background/Objectives: Omicron, the predominant variant of SARS-CoV-2, exhibits strong immune-evasive properties, leading to the reduced efficacy of existing vaccines. Consequently, the development of versatile vaccines is imperative. Intranasal mRNA vaccines offer convenient administration and have the potential to enhance mucosal immunity. However, delivering vaccines via the nasal mucosa requires overcoming complex physiological barriers. The aim of this study is to modify PEGylated lipids to enhance the mucosal immune efficacy of the vaccine. Methods: The PEGylated lipid component of lipid nanoparticle (LNP) delivery vectors was modified with chitosan or mannose to generate novel LNPs that enhance vaccine adhesion or targeting on mucosal surfaces. The impact of the mRNA encoding the receptor-binding domain of Omicron BA.4/BA.5 on the immune response was examined. Results: Compared to the unmodified LNP group, the IgG and IgA titers in the chitosan or mannose-modified LNP groups showed an increasing trend. The chitosan-modified group showed better effects. Notably, the PEGylated lipid with 1.5 mol% of chitosan modification produced high levels of IgG1 and IgG2a antibodies, promoting Th1/Th2 responses while also generating high levels of IgA, which can induce stronger cellular immunity, humoral immunity, and mucosal immunity. Conclusions: The 1.5 mol% of chitosan-modified LNPs (mRNA-LNP-1.5CS) can serve as a safe and effective carrier for intranasal mRNA vaccines, offering a promising strategy for combating the Omicron variant.

Predictors of the effectiveness to first-line CTLA4-Ig in patients with rheumatoid arthritis: the FIRST registry.

This study aimed to elucidate which bio-naïve patients with rheumatoid arthritis (RA) are suitable for treatment with CTLA4-Ig. This study enrolled 953 patients with RA who were administered their first biological disease-modifying antirheumatic drug (CTLA4-Ig, n = 328; tumour necrosis factor inhibitor [TNFi], n = 625) from July 2013 to August 2022. The primary outcome was the Clinical Disease Activity Index (CDAI) remission rate at week 24 in each group, adjusted using Propensity Score-based Inverse Probability of Treatment Weighting (PS-IPTW). After minimizing selection bias using PS-IPTW, the CDAI remission showed no significant difference between the CTLA4-Ig and TNFi groups (p= 0.464). Multivariable logistic regression analysis identified low baseline Health Assessment Questionnaire-Disability Index (HAQ-DI) scores as a contributing factor to the CDAI remission rate at week 24 in both groups, along with high baseline anti-citrullinated peptide antibody (ACPA) levels in the CTLA4-Ig group. However, among patients with high baseline HAQ-DI scores and low baseline ACPA levels (≦57.2), the CDAI remission rate was significantly higher in the TNFi group (29.8%) compared with the CTLA4-Ig group (5.9%, p< 0.0001). Among patients with high baseline HAQ-DI scores and ACPA levels (>57.2), the CDAI remission rate was significantly higher in the CTLA4-Ig group (35.6%) compared with the TNFi group (22.1%, p= 0.0057). Bio-naive RA patients with low HAQ-DI scores showed high treatment efficacy with no significant difference between CTLA4-Ig and TNFi. Among patients with high baseline HAQ-DI scores, TNFi and CTLA4-Ig were more likely to be effective in those with lower and higher baseline ACPA levels, respectively.

A single dose recombinant AAV based CHIKV vaccine elicits robust and durable protective antibody responses in mice.

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that is responsible for Chikungunya fever, which is characterized by fever, rash, and debilitating polyarthralgia. Since its re-emergence in 2004, CHIKV has continued to spread to new regions and become a severe health threat to global public. Development of safe and single dose vaccines that provide durable protection is desirable to control the spread of virus. The recombinant adeno-associated virus (rAAV) vectors represent promising vaccine platform to provide prolonged protection with a single-dose immunization. In this study, we developed a rAAV capsid serotype 1 vector based CHIKV vaccine and evaluated its protection effect against CHIKV challenge. The recombinant AAV1 encoding the full-length structural proteins of CHIKV (named as rAAV1-CHIKV-SP) was generated in vitro by transfecting the plasmids of AAV helper-free system into HEK-293T cells. The safety and immunogenicity of rAAV1-CHIKV-SP were tested in 4-week-old C57BL/6 mice. The antibody responses of the mice receiving prime-boost or single-dose immunization of the vaccine were determined by ELISA and plaque reduction neutralizing test. The immunized mice were challenged with CHIKV to evaluate the protection effect of the vaccine. The rAAV1-CHIKV-SP showed remarkable safety and immunogenicity in C57BL/6 mice. A single dose intramuscular injection of rAAV1-CHIKV-SP elicited high level and long-lasting antibody responses, and conferred complete protection against a heterologous CHIKV strain challenge. These results suggest rAAV1-CHIKV-SP represents a promising vaccine candidate against different CHIKV clades with a simplified immunization strategy.

An immuno-inflammatory profiling of asymptomatic individuals in a malaria endemic area in Uganda

Malaria caused by Plasmodium falciparum leads to the destruction of red blood cells (RBCs). A better understanding of how naturally immune individuals control infections should be valuable for future vaccine studies. Antibodies against RBCs and RBC surface antigens were measured together with different inflammatory markers in healthy adults living in a malaria endemic area of Uganda and compared to Swedish healthy adults. Antibodies binding to RBCs were clearly elevated in Ugandans compared to Swedish samples, and for RBC surface antigens the Ugandans had higher levels of antibodies against JMH, but not against Cromer or Kell. Twenty-eight percent of the Ugandans were PCR-positive for P. falciparum, and these had higher levels of IgG against parasite extract and more inhibition in functional growth/invasion assays, but levels of antibodies against RBC, RBC surface antigens, results from Direct Antiglobulin Tests (DAT) and indirect antiglobulin tests were similar when compared with PCR-negative individuals. When inflammatory markers (α-1-antitrypsin, haptoglobin, orosomucoid/α-1-acid glycoprotein, CRP, IgG, IgA and IgM) were measured there were in general almost no signs of inflammation except for clearly elevated levels of IgG. Some had low levels of haptoglobin and for orosomucoid more than half of the individuals had clearly reduced levels. There was no correlation between the inflammatory markers and PCR-positivity, antibodies against RBCs or parasites. In conclusion, for healthy adults living in a malaria endemic area, there was a clear presence of antibodies against RBCs in parallel with high levels of IgG and almost no signs of inflammation, even though many individuals were carrying parasites.

Toxoplasmosis infection in an HIV-negative patient presenting with clinical and MRI findings similar to those of multiple sclerosis

Toxoplasmosis is a parasitic infection that can present in various clinical forms, ranging from asymptomatic to severe neurological manifestations. The primary sources of infection include undercooked meat, unwashed produce and contact with cat faeces. Toxoplasmosis can lead to encephalitis, particularly in immunocompromised patients, and is often misdiagnosed as other neurological conditions such as multiple sclerosis (MS). We report the case of a 44-year-old male from Almaty, Kazakhstan, who presented with neurological symptoms including headaches, dizziness, diplopia, leg weakness and elevated blood pressure. The patient had a history of consuming undercooked meat, but no prior neurological conditions. Initial magnetic resonance imaging (MRI) revealed demyelinating lesions, leading to a diagnosis of MS. However, high levels of IgG antibodies against Toxoplasma gondii were detected, prompting further testing. A polymerase chain reaction test for toxoplasmosis was negative, but the patient was treated empirically with trimethoprim and sulfamethoxazole for six months. A follow-up MRI showed a significant reduction in brain lesions, and the patient’s symptoms improved.

Clinical value of anti-DSG2 antibodies in arrhythmogenic right ventricular cardiomyopathy: a real-life experience

Abstract Background/Introduction Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited myocardial disease leading to ventricular arrhythmias and heart failure. The diagnosis is challenging and currently based on a set of clinical criteria with limited sensitivity and specificity. Anti-desmoglein-2 (DSG2) antibodies were previously found in patients diagnosed with ARVC, while they were absent in both healthy controls and athletes. Purpose The study aimed to evaluate the diagnostic value of anti-DSG2 antibodies by comparing their concentration in patients with ARVC and other right ventricular diseases. Additionally, the prognostic value of the biomarker was assessed. Methods A cohort of 101 patients with a definite diagnosis of ARVC according to the 2010 Task Force criteria was enrolled (64 males, mean age 47 ± 16 years). The control group included patients with Ebstein anomaly and Eisenmenger syndrome (16 and 21 subjects, respectively). Anti-DSG2 antibody concentration was assessed using enzyme-linked immunosorbent assay (ELISA). After that, an ARVC cohort was followed for 3.9 ± 1.6 years for the occurrence of the primary endpoint of cardiac death or heart transplantation (HTx) and major arrhythmic events (MAEs) defined as ventricular fibrillation, sustained ventricular tachycardia (sVT) or appropriate implantable cardioverter-defibrillator intervention. Results The median anti-DSG2 antibody concentration presented as ELISA optical density (OD) reached 1.36 [0.54-2.15] in the ARVC cohort, while it was 0.4 [0.34-0.54] in Ebstein anomaly group and 0.36 [0.28-0.58] in the Eisenmenger syndrome group. There was a significant difference in antibody levels between ARVC and both control groups (p &amp;lt; 0.001, Figure 1). The cutoff value for the diagnosis of ARVC was 0.774, with 72.3% sensitivity and 100% specificity (AUC = 0.823). After dividing the ARVC cohort according to the designated cutoff point, the anti-DSG2-positive subgroup was significantly younger (44.5 ± 15.4 vs. 52.8 ± 15.5 years, p = 0.017) and more likely to have a history of sVT (68.5% vs. 46.4%, p = 0.041). There was no difference regarding sex, presence of PKP2 or DSG2 mutations, previous sports activities, history of sudden cardiac arrest or syncope, right ventricular dimensions and systolic function, left ventricular ejection fraction and heart failure symptoms. This threshold was not predictive for the primary endpoint and MAEs. However, a more stringent cutoff value for anti-DSG2 level (1.021) was found to predict death or HTx (HR [95% CI] 4.56 [1.05-19.76], p = 0.026, Figure 2). Conclusions The concentration of anti-DSG2 antibodies is significantly higher in ARVC compared to other right ventricular diseases. A high antibody level is prognostic for death or HTx in ARVC.Figure 1Figure 2

Knowing the Antibody Status may influence compliance of health care workers to COVID-19 booster dose

Abstract Background Previous studies showed that the fourth SARS-CoV-2 vaccine dose has a protective effect against infection, as well as against severe disease and death. This study aimed to examine whether knowledge of a high level antibody after the 3rd dose may reduce compliance to the 4th booster dose among health care workers. Methods We conducted a prospective cohort study among health care workers vaccinated with the first three doses at Rambam Healthcare Campus, a tertiary hospital in northern Israel. Participants underwent a serological test before the 4th booster vaccine was offered to all of them, with results provided to participants. The population was divided into two groups: those with antibody below 955 AU/ml, and those with 955 AU/ml and higher, a cutoff found protective in a previous study. Multiple logistic regression was carried out to compare the compliance to the 4th booster between the two groups, adjusted for demographic and clinical variables. Results After adjusting for the confounding variables, the compliance was higher in those with antibody levels below 955 AU/ml (OR = 1.41, P = 0.05, 95% CI 1.10-1.96). In addition, male sex, and age of 60 years and above were also associated with higher vaccination rate (OR = 2.28, P &amp;lt; 0.001, 95% CI 1.64 - 3.17), (OR = 1.14, P = 0.043, 95% CI 1.06 - 1.75), respectively. Conclusions Knowledge of the antibody status may affect compliance with the booster dose. Considering waning immunity over time, reduced compliance may affect the protection of health care workers who declined the 4th dose. Key messages • Knowing the antibody status may affect compliance of health care workers with the booster dose. • Reduced compliance of health care workers may affect their protection.

Evaluation of spexin levels in euthyroid patients with Hashimoto thyroiditis and its relation to autoimmunity.

Hashimoto thyroiditis (HT) is chronic lymphocytic thyroiditis. Cytokines and chemokines such as tumor necrosis factor-alpha, interferon-gamma, and interleukin-1 beta originating from immune cells are involved in the etiopathogenesis of HT. Spexin (SPX) is a recently identified novel peptide hormone consisting of 14 amino acids and has been demonstrated in follicle epithelial cells in thyroid tissue. SPX has been shown to affect the inflammatory response and play a role in its regulation in various diseases. There is a need for markers for diagnosis and treatment of HT patients with negative antibody levels. We found that there is no study in the literature that investigates the HT and the role of spexin in this inflammatory process. Forty-five patients aged 18 to 70 years with HT or newly diagnosed HT and 42 healthy subjects as the control group were included in the study. Patients in the HT group were divided into 3 categories according to ultrasound findings. Mild heterogeneity was called grade 1 (G1), moderate heterogeneity was called grade 2 (G2), and high heterogeneity was called grade 3 (G3). Laboratory parameters and anthropometric measurements of all patients participating in the study were performed, and SPX was measured by the ELISA method. There was no significant difference between the HT and control groups in terms of SPX levels (P = .27). In HT subgroup analysis, SPX levels were found to be borderline statistically significantly higher in the G2 group, where antibody levels were higher compared to other groups (P = .061). In our study, we evaluated SPX levels in HT patients, which has never been done before in the literature. We found high SPX levels in HT patients with high antibody levels. Multicenter studies with high case series, especially at the tissue level, are needed to fully explain the role of SPX in HT immunoetiopathogenesis and to understand immune-checkpoint pathways more clearly.

Improved influenza vaccine responses after expression of multiple viral glycoproteins from a single mRNA.

Influenza viruses cause substantial morbidity and mortality every year despite seasonal vaccination. mRNA-based vaccines have the potential to elicit more protective immune responses, but for maximal breadth and durability, it is desirable to deliver both the viral hemagglutinin and neuraminidase glycoproteins. Delivering multiple antigens individually, however, complicates manufacturingand increases cost, thus it would be beneficial to express both proteins from a single mRNA. Here, we develop an mRNA genetic configuration that allows the simultaneous expression of unmodified, full-length NA and HA proteins from a single open reading frame. We apply this approach to glycoproteins from contemporary influenza A and B viruses and, after vaccination, observe high levels of functional antibodies and protection from disease in female mouse and male ferret challenge models. This approach may further efforts to utilize mRNA technology to improve seasonal vaccine efficacy by efficiently delivering multiple viral antigens simultaneously and in their native state.

Evaluation of Antibody Response and Reinfection in Improved Hypertensive, Cardiac, and Diabetic Patients with COVID-19 for Six Months in Unvaccinated Outpatients

Background: This study presents a unique investigation into the relationship between the persistence of IgG antibody levels and reinfection in recovered coronavirus disease 2019 (COVID-19) patients. By shedding light on this aspect, we aim to address concerns about the risk of reinfection in unvaccinated patients, offering a fresh perspective in the current COVID-19 research landscape. Objectives: This study takes a novel approach by focusing on the antibody response and reinfection in COVID-19 patients who were treated in outpatient settings for six months. This approach distinguishes our research within the broader context of COVID-19 studies. Methods: Our comprehensive and rigorous prospective cohort study included 149 COVID-19 outpatients referred to Imam Ali Clinic in Dezfoul City, Khuzestan province, Iran. Data was meticulously collected from September 2021 to February 2022. All unvaccinated patients underwent RT-PCR testing, and their anti-SARS-CoV-2 IgM/IgG levels were measured at the onset of illness. Serum IgG levels were then measured again after three and six months using ELISA. Results: All COVID-19 patients had their IgG antibody levels tested three times (on days two to four of the illness, at three months, and at six months). Significant differences in IgG antibody levels were observed between the vaccinated and unvaccinated groups during each period (P &lt; 0.05). Additionally, there were substantial differences between the hypertension and non-hypertension groups (P = 0.032). Importantly, our findings reaffirm the impartiality of our research, as no significant differences were found between reinfection rates and various demographic or disease-related factors. Conclusions: The findings of this study suggest that high levels of IgG antibodies in unvaccinated patients could potentially reduce the risk of reinfection. This insight could have important implications for public health strategies and the management of COVID-19 in unvaccinated populations, providing valuable information for decision-makers in the field.

Herpesvirus Antibody Response and Occurrence of Symptoms in Acute and Post-Acute COVID-19 Disease.

Knowledge about the underlying causes of the individual occurrence of symptoms during acute COVID-19 disease and during the post-acute sequelae of COVID-19 is limited. In a German COVID-19 follow-up study, we assessed whether elevated antibody responses to herpesviruses were associated with symptom occurrence in acute COVID-19 disease (n = 96 participants) and during 20 months of follow-up (n = 62 participants). Serum samples were analyzed for their antibodies to herpes simplex virus (HSV)-1 and -2, Epstein-Barr virus (EBV), and Cytomegalovirus (CMV) using fluorescent bead-based multiplex serology. The association of herpesvirus antibodies with symptom occurrence (fatigue, fever, dyspnea, decrease in taste, concentration problems) was assessed using multivariate logistic regression models. High EBV antibody levels were significantly associated with a more than fourfold increased odds of experiencing fatigue during acute COVID-19 disease and during follow-up. High CMV antibody levels were significantly associated with a more than threefold increased odds of experiencing concentration problems and a decrease in taste during the follow-up. The HSV-1 and -2 antibody levels were not elevated in the individuals that experienced symptoms. In conclusion, our findings indicate that herpesvirus infections, specifically EBV and CMV infections, might play a role in symptom development during acute and post-acute COVID-19 disease. It remains to be elucidated whether the elevated EBV and CMV antibodies determined in our study are indicators of herpesvirus reactivation.

6674 Non-Progressive Diabetes Leading to Neurologic Diagnosis

Abstract Disclosure: F.L. Thelmo: None. M. Murugesh: None. B. Simon: None. Introduction: The autoantibody GAD (anti-glutamic acid decarboxylase) is highly associated with Type 1 diabetes (T1DM) and Latent Autoimmune Diabetes in Adulthood (LADA). If GAD is detected before the onset of overt diabetes, typically 80% of patients with abnormal glucose tolerance progress to insulin deficiency and the need for insulin treatment after 5 years.[1] Stiff Person Syndrome (SPS) is also associated with GAD antibodies, but in SPS, the GAD antibodies are believed to be different epitopes that target GABA production in neuromuscular pathways. We present a case of presumed LADA that did not progress for 30 years, with progressive neurologic symptoms leading to a diagnosis of SPS in later adulthood.Case: A 52-year-old female presented for evaluation of T1DM and worsening neuropathies. She had a long history of prediabetes, with mildly elevated glucose values noted on labs since her early twenties. There was no family history of T1DM or autoimmune conditions. Six years ago, she sought endocrine evaluation. HgbA1c values were 6.1 to 6.6%. Intermittent use of continuous glucose monitoring (CGM) revealed post-prandial hyperglycemia up to 180mg/dL, occasionally (but rarely) exceeding 200mg/dL. She recalls being told that she had positive antibodies (unclear which ones tested) and given a diagnosis of T1DM. She declined insulin and maintained glycemic control with carbohydrate restriction and exercise; there were no episodes of diabetic ketoacidosis. She then developed neurologic symptoms: fullness, tightness and bloating in the abdomen (diagnosed with gastroparesis on scanning) and after a COVID 19 booster in 2021, muscle spasms in the legs and back with worsening abdominal tightness. At the time of her current presentation, A1C was 6.4%, glycated albumin was 14.4% (prediabetes range 13.6 - 15.5%) and CGM time in range was 99%. A non-fasting C-peptide was 3.6 ng/mL (1.1- 4.4ng/mL) with glucose of 119 mg/dL. Repeat antibody testing revealed undetectable IA-2 and ZnT8 antibodies, but GAD was grossly elevated at 1,015.3 U/mL. Given the unusually high GAD titer, SPS was suspected, and this diagnosis was confirmed on referral to neurology. The patient was advised to liberalize diet and will be monitored with HgbA1c and serial C-peptide levels to assess progression in the future. Discussion: While high GAD antibody levels are associated with progression to insulin dependence in adults with LADA, in SPS the GAD titers tend to be even higher2. The presence of only one auto-antibody for T1DM (and not multiple), long-standing non-progressive hyperglycemia, and normal C-peptide pointed away from autoimmune diabetes being the etiology for mild glucose intolerance in this patient. If neurologic symptoms are present, high GAD should prompt the evaluation for other syndromes associated with GAD antibody epitopes such as SPS. Presentation: 6/2/2024

Development of a fully automated chemiluminescent immunoassay for the quantitative and qualitative detection of antibodies against African swine fever virus p72.

African swine fever (ASF), caused by ASF virus (ASFV), is a highly infectious and severe hemorrhagic disease of pigs that causes major economic losses. Currently, no commercial vaccine is available and prevention and control of ASF relies mainly on early diagnosis. Here, a novel automated double antigen sandwich chemiluminescent immunoassay (DAgS-aCLIA) was developed to detect antibodies against ASFV p72 (p72-Ab). For this purpose, recombinant p72 trimer was produced, coupled to magnetic particles as carriers and labeled with acridinium ester as a signal trace. Finally, p72-Ab can be sensitively and rapidly measured on an automated chemiluminescent instrument. For quantitative analysis, a calibration curve was established with a laudable linearity range of 0.21 to 212.0 ng/mL (R2 = 0.9910) and a lower detection limit of 0.15 ng/mL. For qualitative analysis, a cut-off value was set at 1.50 ng/mL with a diagnostic sensitivity of 100.00% and specificity of 98.33%. Furthermore, antibody response to an ASF gene-deleted vaccine candidate can be accurately quantified using this DAgS-aCLIA, as evidenced by early seroconversion as early as 7 days post-immunization and high antibody levels. Compared with available enzyme-linked immunosorbent assays, this DAgS-aCLIA demonstrated a wider linearity range of 4 to 16-fold, and excellent analytical sensitivity and agreement of over 95.60%. In conclusion, our proposed DAgS-aCLIA would be an effective tool to support ASF epidemiological surveillance.IMPORTANCEAfrican swine fever virus (ASFV) is highly contagious in wild boar and domestic pigs. There is currently no vaccine available for ASF, so serological testing is an important diagnostic tool. Traditional enzyme-linked immunosorbent assays provide only qualitative results and are time and resource consuming. This study will develop an automated chemiluminescent immunoassay (CLIA) that can quantitatively and qualitatively detect antibodies to ASFV p72, greatly reducing detection time and labour-intensive operation, and improving detection sensitivity and linearity range. This novel CLIA would serve as a reliable and convenient tool for ASF pandemic surveillance and vaccine development.

A longitudinal study of the dynamics of Mycoplasma bovis antibody status in primiparous cows and bulk tank milk in Swedish dairy herds

Mycoplasma (M.) bovis is an important pathogen causing pneumonia, mastitis, and arthritis in cattle all over the world entailing reduced animal welfare and economic losses. In this longitudinal study, we investigated the presence of M. bovis antibodies in bulk tank milk (BTM) and in milk from primiparous (PP) cows at 4 sampling occasions over 2 years. Herd characteristics associated with a positive antibody test result in PP cows were investigated. The participating dairy herds (n = 149) were situated in southern Sweden, samples were collected and analyzed with ID Screen antibody ELISA. Information on herd characteristics was retrieved from the national Dairy Herd Improvement database. To identify herd characteristics associated with the presence of antibodies in PP cows, mixed linear regression with herd and sample as random factors were used. The apparent herd-level prevalence of M. bovis infection based on antibodies in BTM was 17% but with the addition of PP cows the prevalence increased to 28%. The results showed that larger herds and introduction of cattle was associated with higher antibody levels in PP cows. In conclusion, this study showed a clear difference in the apparent prevalence of M. bovis infection based on antibodies in BTM or in PP cows, the no. of positive herds were almost doubled when including PP cows. This motivates repeated sampling of a few PP cows to find newly infected herds in an early stage. Finally, the results showed that introduction of cattle influences the level of M. bovis antibodies. This is important in the control and prevention of further spread of the infection. It is essential for free herds to know their M. bovis status and antibody testing is highly recommended if introducing cattle.

Evaluation The Role Of Some Immunological Markers In Coronavirus-19 Infection In A Sample Of Iraqi Patients

Background: cells produce proinflammatory cytokines like il-8, which activates neutrophils and stimulates immunological responses. Il-6 is produced quickly and transiently in response to infections and stimulates hematopoiesis and acute phase responses, aiding host defense. Dysregulated il-6 production can lead to chronic inflammation and autoimmunity despite its transcriptional and posttranscriptional regulation. Objective: the study aims to identify immunological markers that could serve as diagnostic factors for covid-19 infections, using rt-pcr for diagnosis and estimation of interleukin-6 and interleukin-8 levels and evaluating their association with infection progression, given the importance of covid-19 in various medical fields. Methods: blood samples collected from al-zafaraniyah general hospital, fatima-alzahra hospital, and al-yarmook teaching hospital in baghdad city identified 40 patients with coronavirus. The study used the enzyme-linked immune sorbent assay (elisa) approach to detect interleukin-6 and interleukin-8 antibodies in the patients ' serum. Results: the study reveals a strong correlation between immunological markers and corona virus-19 infection, with 40 patients showing higher levels of interleukin-6 and interleukin-8 antibodies, indicating a significant association between these markers. Conclusions: the study found a significant association between coronavirus-19 infection patients and immunological markers, with high levels of interleukin-6 and interleukin-8 antibodies in serum highly associated with coronavirus -19 incidence.

Seroprevalence of SARS-CoV-2 antibodies in children with nephrotic syndrome and chronic kidney disease: a cross-sectional study from India.

There is a paucity of literature on the seroprevalence of SARS-CoV-2 antibodies among pediatric patients with underlying kidney disorders; few serosurveys among healthy children have shown seropositivity of 20-65% after different waves of infections. The study had a cross-sectional design and was conducted between January 2023 and July 2023; 163 children and adolescents (1-18years) with nephrotic syndrome and chronic kidney disease (CKD) were screened for Anti-Spike SARS-COV-2 IgG antibodies as detected by a quantitative chemiluminescence immunoassay. Children with nephrotic syndrome, both steroid sensitive (SSNS) and steroid resistant (SRNS) were enrolled during disease remission. Correlation of SARS-CoV-2 seropositivity status was done with age, gender, disease type, treatment duration, immunosuppressants, previous SARS-CoV-2 infection, and immunization status. Of 163 children (63.8% boys) with median age of 9years; 101 (62%) had underlying nephrotic syndrome (61 SSNS and 40 SRNS), and 62 (38%) children had CKD. Seroprotective titers for SARS-COV2 antibodies were present in 100 (61.3%) children. The median titers for all patients were 37.1 BAU/mL; for nephrotic syndrome they were 27.1 BAU/mL and for CKD they were 76.7 BAU/mL (p = 0.0033). A total of 43 (26.4%) children had high positive antibody levels (> 200 BAU/ml). Among those with nephrotic syndrome 60.7% with SSNS and 43.5% SRNS had seropositive titers. Only 4 (2.5%) children had a history of previous COVID infection and 6 (3.7%) were vaccinated. In a largely unvaccinated population of children with nephrotic syndrome and CKD, 61.3% were seropositive for SARS-CoV-2 IgG antibody indicating a past asymptomatic infection; titers were significantly higher in CKD compared to nephrotic syndrome.

Constructing the cGAMP-Aluminum Nanoparticles as a Vaccine Adjuvant-Delivery System (VADS) for Developing the Efficient Pulmonary COVID-19 Subunit Vaccines.

The cGAMP-aluminum nanoparticles (CAN) are engineered as a vaccine adjuvant-delivery system to carry mixed RBD (receptor-binding domain) of the original severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its new variant for developing bivalent pulmonary coronavirus disease 2019 (COVID-19) vaccines (biRBD-CAN). High phosphophilicity/adsorptivity made intrapulmonary CAN instantly form the pulmonary ingredient-coated CAN (piCAN) to possess biomimetic features enhancing biocompatibility. In vitro biRBD-CAN sparked APCs (antigen-presenting cells) to mature and make extra reactive oxygen species, engendered lysosome escape effects and enhanced proteasome activities. Through activating the intracellular stimulator of interferon genes (STING) and nucleotide-binding domain and leucine-rich repeat and pyrin domain containing proteins 3 (NALP3) inflammasome pathways to exert synergy between cGAMP and AN, biRBD-CAN stimulated APCs to secret cytokines favoring mixed Th1/Th2 immunoresponses. Mice bearing twice intrapulmonary biRBD-CAN produced high levels of mucosal antibodies, the long-lasting systemic antibodies, and potent cytotoxic T lymphocytes which efficiently erased cells displaying cognate epitopes. Notably, biRBD-CAN existed in mouse lungs and different lymph nodes for at least 48h, unveiling their sustained immunostimulatory activity as the main mechanism underlying the long-lasting immunity and memory. Hamsters bearing twice intrapulmonary biRBD-CAN developed high resistance to pseudoviral challenges performed using different recombinant strains including the ones with distinct SARS-CoV-2-spike mutations. Thus, biRBD-CAN as a broad-spectrum pulmonary COVID-19 vaccine candidate may provide a tool for controlling the emerging SARS-CoV-2 variants.

A cynomolgus monkey E. coli urinary tract infection model confirms efficacy of new FimH vaccine candidates.

The increase in urinary tract infections (UTI) caused by antibiotic-resistant Escherichia coli requires the development of new therapeutic agents and prophylactic vaccines. To evaluate the efficacy of new lead candidates, we implemented a cynomolgus macaque UTI challenge model that mimics human uncomplicated cystitis in response to transurethral challenge with a multidrug-resistant (MDR) E. coli serotype O25b ST131 isolate. E. coli fimbrial adhesin FimH and O-antigens are separately under clinical evaluation by others as vaccine candidates to prevent UTI and invasive urosepsis disease, respectively. Accordingly, we assessed the protective efficacy of three 50-µg intramuscular doses of a novel recombinant FimH antigen adjuvanted with liposomal QS21/MPLA compared with saline placebo in groups of nine animals. A third group was vaccinated with this FimH formulation in combination with 1 µg each of a four-valent mixture of serotype O1a, O2, O6, and O25b O-antigen CRM197 lattice glycoconjugates. Both vaccines elicited high levels of serum FimH IgG and adhesin blocking antibodies at the time of bacterial challenge and, for the combination group, O-antigen-specific antibodies. Following bacterial challenge, both vaccinated groups showed >200- and >700-fold reduction in bacteriuria at day 2 and day 7 post-infection compared with placebo, respectively. In parallel, both vaccines significantly reduced levels of inflammatory biomarkers IL-8 and myeloperoxidase in the urine at day 2 post-infection relative to placebo. Results provide preclinical proof-of-concept for the prevention of an MDR UTI infection by these new vaccine formulations.

Catastrophic Antiphospholipid Syndrome: A Review of Current Evidence and Future Management Practices.

Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by blood clots and pregnancy complications due to antiphospholipid antibodies. Catastrophic APS (CAPS), a severe variant, leads to multiorgan failure and is often fatal. Pathogenesis involves antiphospholipid antibodies, particularly anti-beta-2-glycoprotein I (aβ2GPI), which trigger endothelial cell (EC) activation, cytokine release, and a prothrombotic state. Infections, surgeries, and other triggers can precipitate CAPS, leading to widespread microthromboses and systemic inflammatory responses. CAPS predominantly affects younger patients and those with systemic lupus erythematosus (SLE), with a high mortality rate, though recent treatment advances have improved survival. Diagnosing CAPS involves identifying clinical manifestations, including rapid organ involvement and small vessel occlusions, confirmed by histopathology and high antiphospholipid antibody levels. The CAPS registry data indicate that commonly affected organs include kidneys, lungs, central nervous system, and the heart, with a high prevalence of lupus anticoagulant and anticardiolipin antibodies (aCL). Current management strategies focus on therapeutic anticoagulation, immunosuppressive therapies like corticosteroids, and adjunct treatments such as plasmapheresis and intravenous immunoglobulin (IVIG). Early use of glucocorticoids and combination therapy has significantly improved outcomes. In life-threatening cases, especially with microangiopathy, experts recommend performing plasma exchange (PE). Patients with associated autoimmune conditions or refractory cases may receive cyclophosphamide (CY) and rituximab while considering PE for treatment. Maintenance of anticoagulation with an appropriate international normalized ratio (INR) is crucial to prevent recurrence.This article reviews the pathogenesis and epidemiology of CAPS. It also examines the current management strategies, and discusses the challenges and controversies associated with these strategies. It hereafter offers recommendations for future management and outlines directions for further research.