Presence Of Specific Antibodies Research Articles

Seroepidemiological study of Epstein-Barr virus infection in reproductive age women in Bulgaria

Abstract Background At the present stage, studies on the involvement of Epstein-Barr virus (EBV) in neonatal pathology are rare. Establishing seroprevalence among women of reproductive age has important epidemiological significance in order to determine the risk of infection during pregnancy and intrauterine infection of the fetus. The aim of the study is to determine the seroepidemiological status of EBV in reproductive age women. Methods We conducted a prospective seroepidemiological study in which 96 healthy women of reproductive age (18 to 49 years) were tested during the period December 2023 - January 2024 in Medical Center “Clinical Institute for Reproductive Medicine”- Pleven, Bulgaria. Participants were surveyed on a voluntary basis. The exclusion criteria were: presence of autoimmune disorder, immunosuppressive states, malignancy and populations at risk such as persons under 18 years of age and pregnant women. The presence of specific antibodies were detected using a standardized Anti-EBV VCA IgM, Anti-EBV VCA IgG and Anti-EBV EBNA-1 IgG ELISA kits. The demographic and anamnestic data were collected for each participant in Case Report Form. Results The average age of the women was 34.41±5.44 years. Dominating were women with higher education (76.04%) living in urban areas (91.67%). The results of the seroepidemiological study of markers for EBV show that 98.96% of women of reproductive age are seropositive. There was no significant correlation between the presence of antibodies and socio-demographic indicators (living area, education and number of family members). Conclusions The results of this study imply that only 1.04% of the reproductive age women in Pleven region are susceptible to infection with EBV. Although interpretation of the serological status indicates a low risk of primary infection, active monitoring of high-risk groups pregnant women is necessary because of the possibility of EBV reactivation. Key messages • On-going monitoring of the seroepidemiological EBV status in reproductive age women is necessary to determining the possibility of EBV reactivation and to reduce the teratogenic risk of the infection. • Seroepidemiological surveillance of markers for EBV infection in reproductive age women is part of effective preventive measures at the public health level.

Optimizing sheep B-cell epitopes in Echinococcus granulosus recombinant antigen P29 for vaccine development.

Echinococcus granulosus is a widespread zoonotic parasitic disease, significantly impacting human health and livestock development; however, no vaccine is currently available for humans. Our preliminary studies indicate that recombinant antigen P29 (rEg.P29) is a promising candidate for vaccine. Sheep were immunized with rEg.P29, and venous blood was collected at various time points. Serum was isolated, and the presence of specific antibodies was detected using ELISA. We designed and synthesized a total of 45 B cell monopeptides covering rEg.P29 using the overlap method. ELISA was employed to assess the serum antibodies of the immunized sheep for recognition of these overlapping peptides, leading to the preliminary identification of B cell epitopes. Utilizing these identified epitopes, new single peptides were designed, synthesized, and used to optimize and confirm B-cell epitopes. rEg.P29 effectively induces a sustained antibody response in sheep, particularly characterized by high and stable levels of IgG. Eight B-cell epitopes of were identified, which were mainly distributed in three regions of rEg.P29. Finally, three B cell epitopes were identified and optimized: rEg.P2971-90, rEg.P29151-175, and rEg.P29211-235. These optimized epitopes were well recognized by antibodies in sheep and mice, and the efficacy of these three epitopes significantly increased when they were linked in tandem. Three B-cell epitopes were identified and optimized, and the efficacy of these epitopes was significantly enhanced by tandem connection, which indicated the feasibility of tandem peptide vaccine research. This laid a solid foundation for the development of epitope peptide vaccine for Echinococcus granulosus.

Detection of small ruminant Lentivirus proviral DNA in red deer from Poland

Small ruminant lentiviruses (SRLVs) are widespread and infect goats and sheep. Several reports also suggest that SRLVs can infect wild ruminants. The presence of specific antibodies against SRLVs has been identified in wild ruminants from Poland, but no studies have been conducted to detect proviral DNA of SRLVs in these animals. Therefore, the purpose of this study was to examine samples from Polish wild ruminants to determine whether these animals can serve as reservoirs of SRLVs under natural conditions. A total of 314 samples were tested from red deer (n = 255), roe deer (n = 52) and fallow deer (n = 7) using nested real-time PCR. DNA from positive real-time PCR samples was subsequently used to amplify a CA fragment (625 bp) of the gag gene, a 1.2 kb fragment of the pol gene and an LTR-gag fragment. Three samples (0.95%) were positive according to nested real-time PCR using primers and probe specific for CAEV (SRLV group B). All the samples were negative for the primers and probe specific for MVV (SRLV A group). Only SRLV LTR-gag sequences were obtained from two red deer. Phylogenetic analysis revealed that these sequences were more closely related to CAEV than to MVV. Our results revealed that deer can carry SRLV proviral sequences and therefore may play a role in the epidemiology of SRLVs. To our knowledge, this is the first study describing SRLV sequences from red deer.

An image-based real time method to measure antibody-dependent cellular phagocytosis

Abstract Antibody-dependent cellular phagocytosis (ADCP) is an important mechanism of action of therapeutic antibodies. In vitro quantification of ADCP has been technically challenging. Fluorescence microscopy and flow cytometry based assays were traditionally used to measure ADCP activity, these methods were static measurements at certain time point, thus provide no information about engulfment kinetics. Moreover, the flow cytometry assays require lifting the macrophage cells from the labware which may affect macrophage activities. We have developed a real time ADCP assay using a live cell analysis system (IncuCyte). The target cells labeled with a pH sensitive dye only fluoresce when they are engulfed in acidic phagosomes of macrophages in the presence of specific antibodies. Our assay is a plate-based with a simple mix and read protocol without disturbing the macrophages. The new method is quantitative and robust, can be readily used for high-throughput screening for functional antibodies. By studying the ADCP activities in different subtypes of macrophages, we found M2 microphages showed the strongest ADCP activity compare to M0 and M1 macrophages, due to their different expression levels of FcgRs. The assay is highly sensitive to differentiate the antibody variants with enhanced Fc function. It can also be used to study the kinetics and mechanisms of ADCP.

Abstract 2059: An image-based real time method to measure antibody-dependent cellular phagocytosis

Abstract Antibody-dependent cellular phagocytosis (ADCP) is an important mechanism of action of therapeutic antibodies. In vitro quantification of macrophage phagocytosis has been technically challenging. Flow cytometry assays were traditionally used for ADCP activity measurements, these methods were static measurements at certain time point, thus provide no information about engulfment kinetics. Moreover, the flow cytometry assays require lifting the macrophage cells from the labware which may affect macrophage activities. We have developed a real time ADCP assay using a live cell analysis system. The target cells labeled with a pH sensitive dye only fluoresce when they are engulfed in acidic phagosomes in the presence of specific antibodies. Our assay is a plate-based with a simple mix and read protocol without disturbing the macrophages. The new method is quantitative and robust, can be readily used for high-throughput screening for functional antibodies. By studying the ADCP activities of different subtypes of macrophages, we found M2 microphages showed the strongest ADCP activity compare to M0 and M1 macrophages, due to their different expression levels of FcgRs. The assay is highly sensitive to differentiate the antibody variants with enhanced Fc function. It can also be used to study the kinetics and mechanisms of ADCP. Citation Format: Jianyong Wang, Yongchang Shi, Yonglian Sun, James T. Koerber, Akiko Seki, Sascha Rutz. An image-based real time method to measure antibody-dependent cellular phagocytosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2059.

Seroprevalence and Risk Factors of Chlamydia Infection in Pigs in Hunan Province, Southern China, 2017-2018.

Background: Chlamydia is a Gram-negative obligate intracellular bacterium that is pathogenic for humans and a large variety of veterinary animal species. However, there is no continuous monitoring of chlamydia infection data in pigs in Hunan province, southern China. Therefore, in order to evaluate the seroprevalence and identify risk factors associated with Chlamydia infection in pigs within this region, a comprehensive study was conducted. Methods: A total of 3848 serum samples were collected from pigs (from farmers and companies) between May 2017 and August 2018. The presence of specific antibodies against Chlamydia was determined through the employment of the indirect hemagglutination assay (IHA). Results: The overall seroprevalence of Chlamydia was determined to be 26.90% (1038/3848, 95% confidence interval: 25.60-28.40). By employing statistical analysis using SPSS software (p < 0.05), factors such as altitude, sampling regions, and rearing systems of pigs were identified as potential risk factors for Chlamydia infection. Conclusion: These findings elucidate a substantial prevalence of Chlamydia in pigs within the mountainous region of Hunan province, southern China, thereby highlighting a potential risk to human health. These results underscore the need for proactive measures and targeted interventions to mitigate the transmission of Chlamydia in porcine populations, safeguarding both animal welfare and public health.

Relationship between clinical manifestations and serological profile in patients affected by Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder characterized by a variety of both signs and symptoms; it mainly affects women of childbearing age, with an estimated prevalence of 24/100,000 people in Europe and North America. SLE is often described as an antibodies-driven disease as its clinical manifestations are usually associated with the presence or the absence of specific antibodies. To evaluate clinical manifestations in patients with SLE and to assess the relationship with the presence of specific antibodies by using real-world data. A retrospective study was performed; the 2019 EULAR/ACR Classification Criteria for Systemic Lupus Erythematosus were used to classify patients with SLE. Data concerning serological profiles (which included Antinuclear antibodies - ANA, anti dsDNA, anti-Ro/SS-A, anti-La/SS-B, anti-Smith) were gathered along with medical records of clinical manifestations. Complement levels were also tested for possible clinical correlations. χ² or Fisher's exact tests were utilized to establish associations between autoantibodies and symptoms. The odds ratios (OR) and their 95% confidence intervals (CI) were computed. No correction was made for multiple testing; only a p-value 0.01 ≤ was considered significant. One-hundred and twenty-seven patients (n=127, mean age 53.43 ± 14.02) were enrolled in this study. Anti-dsDNA antibodies were found to be statistically significant for both malar rash and proteinuria; anti-Ro/SSA antibodies showed an association with photosensitivity and pericarditis; furthermore, a strong association was found between anti-Ro antibodies and proteinuria, but only if anti-dsDNA antibodies were present as well. Patients who tested positive for anti-La/SSB antibodies correlated with a threefold increase in the risk of developing pericarditis. Lastly, anti-Smith appeared to be associated with NPSLE as well as an increased risk for both autoimmune hemolytic anemia and thrombocytopenia. In our study, many associations confirmed those found in previous studies; however, new relationships between antibodies and clinical manifestations were found thus indicating the need for additional evaluations to assess these correlations further.

Сучасні підходи до вакцинації в дітей з різними варіантами первинних та вторинних імунодефіцитів (огляд літератури)

Chronic disease and immunosuppression increase the risk of vaccine-preventable infections, complications, and mortality from infectious diseases. Safe vaccination and achieving an adequate serological response to vaccination are crucial aspects of the management of such patients. Purpose - to analyze data from domestic and foreign medical literature regarding modern approaches to vaccination of children with various primary and secondary immunodeficiency diseases. The majority of children with primary immunodeficiency who have preserved antibody production can demonstrate an immune response to vaccination, but this response can be rapidly lost over time. Limitations to vaccinating children with primary immunodeficiency are related to the use of live vaccines in conditions where the cellular immune system is compromised. Patients with primary antibody deficiency are treated with immunoglobulin preparations and depend on the presence of specific antibodies at a protective level against infectious pathogens, including vaccine-preventable infections. Several studies have shown the effectiveness of intravenous immunoglobulin treatment in reducing the number and severity of infections in patients with humoral deficiency. The majority of children with rheumatic diseases vaccinated according to the Schedule of Routine Immunisations before starting immunosuppressive therapy have protective antibody levels against vaccine-preventable infections. All non-live vaccines can be administered without restrictions, but should be administered two weeks before immunosuppressive therapy to increase immunogenicity. Children receiving high doses of immunosuppressive therapy should avoid receiving live attenuated vaccines. No conflict of interests was declared by the author.

B-Cell Epitopes-Based Chimeric Protein from SARS-CoV-2 N and S Proteins Is Recognized by Specific Antibodies in Serum and Urine Samples from Patients.

The impact of the COVID-19 pandemic caused by the SARS-CoV-2 virus underscored the crucial role of laboratorial tests as a strategy to control the disease, mainly to indicate the presence of specific antibodies in human samples from infected patients. Therefore, suitable recombinant antigens are relevant for the development of reliable tests, and so far, single recombinant proteins have been used. In this context, B-cell epitopes-based chimeric proteins can be an alternative to obtain tests with high accuracy through easier and cheaper production. The present study used bioinformatics tools to select specific B-cell epitopes from the spike (S) and the nucleocapsid (N) proteins from the SARS-CoV-2 virus, aiming to produce a novel recombinant chimeric antigen (N4S11-SC2). Eleven S and four N-derived B-cell epitopes were predicted and used to construct the N4S11-SC2 protein, which was analyzed in a recombinant format against serum and urine samples, by means of an in house-ELISA. Specific antibodies were detected in the serum and urine samples of COVID-19 patients, which were previously confirmed by qRT-PCR. Results showed that N4S11-SC2 presented 83.7% sensitivity and 100% specificity when using sera samples, and 91.1% sensitivity and 100% specificity using urine samples. Comparable findings were achieved with paired urine samples when compared to N and S recombinant proteins expressed in prokaryotic systems. However, better results were reached for N4S11-SC2 in comparison to the S recombinant protein when using paired serum samples. Anti-N4S11-SC2 antibodies were not clearly identified in Janssen Ad26.COV2.S COVID-19-vaccinated subjects, using serum or paired urine samples. In conclusion, this study presents a new chimeric recombinant antigen expressed in a prokaryotic system that could be considered as an alternative diagnostic marker for the SARS-CoV-2 infection, with the potential benefits to be used on serum or urine from infected patients.

Celiac disease in children - diagnosis in light of the current 2020 ESPGHAN guidelines

Celiac disease is a chronic immune disorder of the small intestine that is triggered by the consumption of gluten in genetically predisposed individuals. The disease is difficult to diagnose and treat, and its incidence is increasing. It can have different symptoms and clinical manifestations in individuals. Diagnosis is usually based on the presence of specific antibodies and histopathological examination of biopsy specimens from the small intestine. The only effective treatment is a gluten-free diet for life. Diagnosis of celiac disease in children is mainly based on the presence of specific an- tibodies in the blood, such as antibodies to tissue transglutaminase (tTG), antibodies to endomysium (EMA) or deamidated gliadin peptides (DGP). In addition, genetic testing is also used in diagnosis to detect the presence of HLA DQ2 or DQ8 genes, which are often present in people with celiac disease. The final diagnosis of celiac disease is based on a combination of the results of these tests and the presence of characteristic lesions in the small intestine, which are detected during endoscopy and small bowel biopsy.

Tick-borne encephalitis virus IgG antibody surveillance: vaccination- and infection-induced seroprevalences, south-western Germany, 2021.

BackgroundThe exact epidemiology of tick-borne encephalitis virus (TBEV) infections is unknown because many TBEV infections have an influenza-like or asymptomatic course. Surveillance data are based on patients with any (predominantly neurological) symptoms that prompted diagnostic testing. Infection- and vaccine-induced antibodies against TBEV can be distinguished using an NS1IgG ELISA.AimIn a seroprevalence study we aimed to investigate TBEV antibody prevalence, incidences, manifestation indices and potential protection rates in a highly endemic district in south-western Germany.MethodsWe analysed 2,220 samples from healthy blood donors collected between May and September 2021. The reported number of TBEV infections was provided on a sub-district level by the local public health authorities. Blood samples were first screened using a TBEVIgG ELISA. In a second step, all positive samples were further analysed with a recently established NS1IgG ELISA. The presence of specific antibodies against TBEV (excluding cross-reacting antibodies against other flaviviruses) was confirmed by testing screening-positive samples with a microneutralisation assay.ResultsOf 2,220 included samples, 1,257 (57%) tested positive by TBEVIgG ELISA and 125 tested positive for infection-induced TBEV NS1 antibodies, resulting in a TBEV NS1IgG seroprevalence at 5.6% in our population. The yearly incidence based on the NS1 ELISA findings resulted in 283 cases per 100,000 inhabitants.ConclusionUsing the TBEV NS1IgG assay, we confirmed a manifestation index of ca 2% and a high incidence of predominantly silent TBEV infections (> 250/100,000/year), which exceeds the incidence of notified cases (4.7/100,000/year) considerably.

Incidence of IgA antibodies specific to benzo[a]pyrene and steroid hormones in women with colorectal cancer and breast cancer

Formation of DNA adducts of chemical carcinogens is a trigger for carcinogenesis. Adducts of benzo[a]pyrene metabolites and estradiol metabolites with DNA have been found in normal and tumor cells in healthy women and patients with breast and colorectal cancer. These low-weight compounds in macromolecular complexes induce the synthesis of specific antibodies. Previously, the presence of specific antibodies against benzo[a]pyrene (IgA-Bp), estradiol (IgA-Es) and progesterone (IgA-Pg) was revealed in breast cancer patients. The aim of this study is to identify the putative features of the IgA-Bp, IgA-Es, and IgA-Pg formation in postmenopausal women with colorectal cancer, in comparison with healthy and breast cancer patients. Using a noncompetitive enzyme-linked immunosorbent assay, the content of these antibodies was studied in the blood serum of healthy women (n = 401), patients with colorectal cancer (n = 219) and breast cancer (n = 1469) using conjugates of Bp, Es, and Pg with bovine serum albumin as adsorbed antigens. When compared with healthy people, the patients with colorectal cancer exhibited higher incidence of IgA-Bp 3 (75% vs 37%, p 0.0001, OR = 5.0), as well as more common levels of individual antibody ratios: IgA-Bp/IgA-Es 1 (82% vs 41%, p 0.0001, OR = 6.5); IgA-Bp/IgA-Pg 1.5 (77% vs 20%, p 0.0001, OR = 13.4); IgA-Es/IgA-Pg 1 (89% vs 48%, p 0.0001, OR = 8.7). In breast cancer patients, compared with healthy people, high IgA-Bp values ( 3) were more common (45% vs 37%, p 0.004, OR = 1.4), as well as increased IgA-Bp/IgA-Es ratio 1 (57% vs 41%, p 0.0001, OR = 1.9), IgA-Bp/IgA-Pg 1.1 (71% vs 36%, p 0.0001, OR = 4.4) and IgA-Es/IgA-Pg 1.1 (71% vs 41%, p 0.0001, OR = 3.5). In patients with colorectal cancer, compared with patients with breast cancer we have found higher incidence of increased IgA-Bp values ( 3) (75% vs 45%, p 0.0001), IgA-Es 3 (53% vs 39%, p 0, 0001), and of IgA-Pg 2 (52% vs 44%, p = 0.025), as well as IgA-Bp/IgA- Es 1 (82% vs 57%, p 0.0001, OR = 50.8 ); IgA-Bp/IgA-Pg 1.5 (77% vs 49%, p 0.0001); IgA-Es/IgA-Pg 1.1 (85% vs 71%, p 0.0001). The apparently high serum IgA-Bp levels reflect the formation of DNA-Bp adducts at large scale in target cells in colorectal cancer compared with healthy women and breast cancer patients, due to direct exposure of colon epithelium to Bp from food. Immunoassay for IgA-Bp, IgA-Es and IgA-Pg is proposed for assessing individual risk of colorectal cancer in postmenopausal women. The ratios of IgA Bp/IgA-Pg levels 1.5 represent the most informative marker of individual risk for colorectal cancer.

Use of a Foamy-Virus Vector System to Produce an 'Off-the-Shelf' Fcγ-CR-T Cell Product for the Treatment of Hematological and Solid Tumour Malignancies

Introduction Chimeric antigen receptor (CAR) T cells have transformed the field of cancer immunotherapy. Other approaches, such as the use of Fc gamma chimeric receptor (Fcγ-CR)-T cells have further expanded the applicability of such therapies in both solid and liquid tumours with the added benefit of tackling some of the hurdles associated with CAR-T therapies (such as loss or down-regulation of the target antigen). Previous studies have generated Fcγ-CR-T cells from autologous cells, which carry several limitations (prolonged production time, costly, manufacturing failure), whereas the use of Lentiviral vectors (LV) is endowed with extra limitations (packaging limits and mutagenesis risk). Our group has developed an in-house Fcγ-CR-T cell product, using a safer to LV, foamy virus (FV) vector and T cells from healthy donors. Materials We constructed FV vectors expressing the CD16 V158, which has higher Fc binding and is associated with higher tumour killing, the T cell stimulatory molecule CD3ζ and the costimulatory molecule CD28 and an EFP1a promoter. 2nd generation LV vector backbones were purchased from a commercial vendor. Peripheral blood from healthy individuals were used as T cell sources. T cells were activated by anti-CD3/CD28 beads and transduced with CD16-CR, LV or FV vectors. Transduction efficiency was assayed by flow cytometry (FCM) using an anti-CD16 antibody (Ab), on day 3. The human cell lines Raji, Panc01 and DLD-1 were used for functional assays, in the presence of the Abs Rituximab (anti-CD20) and Cetuximab (anti-EGFR), respectively, at a concentration of 0.1 ug/ml. The CR's Ab-binding capacity was assessed by incubating the CD16-CRs with the Abs for 30mins, followed by an anti-human Ig PE antibody and the related fluorescence intensity was assessed. To determine whether Ab binding to CD16-CR can promote aggregation of effector and target cells, CFSE-labelled target cells were mixed with Ab-coated CD16-CRs for 60 mins and the formation of CFSE-PE doublets was assessed by FCM. Their cytotoxic effect was evaluated against the CFSE-labelled Raji or DLD-1 or Panc01 cells at different ratios (5:1, 10:1) for 18 hours, in the presence of Rituximab (0.1ug/ml) or Cetuximab (0.1ug/ml), respectively. The % of live cells was assessed by flow cytometry and calculated as: [1-live targets (sample)/live targets (control)]x100. Results LV and FV vector titers were between 3-5x10^6 TU/ml and 4-5x10^6 TU/ml, respectively. Transduction efficiency ranged from 58.3-69.2% with FV vectors (MOI 3-5) and 85.2-85.9% with LV vectors (MOI 10-20). The Ab-binding capacity of FV-CD16-CRs was determined to be 68.7% (SEM 2.3) and 71.3% (SEM 3.1), (n=3) for Rituximab and Cetuximab, respectively, whereas that of LV-CD16-CRs was 72.1 (SEM 3.3) and 76.5 (SEM 2.8), (n=3), respectively. Cell aggregation of effector and target cells, assessed as doublets, was: (i) 32%, 39% and 36% for FV-CD16-CRs and (ii) 26%, 31% and 29% for LV-CD16-CRs, coated with Rituximab and Cetuximab, respectively, and it was specific for the Raji, DLD-1 and Panc01 cells, respectively. Next, we assessed whether CD16-CR T cells were able to kill target cells in the presence of specific antibodies. Results showed that the FV-CD16-CR-induced % cell lysis was: (i) 26.3% and 51.5% at 5:1 and 10:1 ratio, respectively, in the presence of Rituximab and Raji cells, (ii) 41.5% and 57.7% at 5:1 and 10:1 ratio, respectively, in the presence of Cetuximab and DLD1 cells and (iii) 39.4% and 57.3% at 5:1 and 10:1 ratio, respectively, in the presence of Cetuximab and Panc01 cells. For LV-CD16-CRs the respective % lysis was comparable. More importantly, this lysis was shown to be specific (no lysis noted with untransduced T cells) and significantly lower in the absence of the antibodies. Conclusion Our group has developed for the first time a FV vector for the generation of CD16-CR T cells, with an efficient gene transfer to human T cells and with potent in vitro cytotoxic properties, similar to their LV-derived counterpart. Overall, we provide a proof of concept that allogeneic, in-house Fcγ-CR-T cells derived from a non-pathogenic viral backbone such as the FV, could be a safe, efficient and affordable alternative to LV-derived vectors for immunotherapy. Our future aim is to further modify these Fcγ-CR-T cells, by interfering with immune checkpoint molecules, in order to achieve better tumour penetration and tackle the anergy induced by the tumour microenvironment.

American College of Gastroenterology Guidelines Update: Diagnosis and Management of Celiac Disease.

This guideline presents an update to the 2013 American College of Gastroenterology Guideline on the Diagnosis and Management of Celiac Disease with updated recommendations for the evaluation and management of patients with celiac disease (CD). CD is defined as a permanent immune-mediated response to gluten present in wheat, barley, and rye. CD has a wide spectrum of clinical manifestations that resemble a multisystemic disorder rather than an isolated intestinal disease, and is characterized by small bowel injury and the presence of specific antibodies. Detection of CD-specific antibodies (e.g., tissue transglutaminase) in the serum is very helpful for the initial screening of patients with suspicion of CD. Intestinal biopsy is required in most patients to confirm the diagnosis. A nonbiopsy strategy for the diagnosis of CD in selected children is suggested and discussed in detail. Current treatment for CD requires strict adherence to a gluten-free diet (GFD) and lifelong medical follow-up. Most patients have excellent clinical response to a GFD. Nonresponsive CD is defined by persistent or recurrent symptoms despite being on a GFD. These patients require a systematic workup to rule out specific conditions that may cause persistent or recurrent symptoms, especially unintentional gluten contamination. Refractory CD is a rare cause of nonresponsive CD often associated with poor prognosis.

JMM Profile: Avian paramyxovirus type-1 and Newcastle disease: a highly infectious vaccine-preventable viral disease of poultry with low zoonotic potential.

Newcastle disease (ND) is a highly contagious disease of poultry caused by virulent avian paramyxovirus-1 (APMV-1) (previously termed avian avulavirus-1 and avian orthoavulavirus-1). APMV-1 is endemic in poultry in many developing countries, whilst outbreaks still occur in developed countries, affecting both commercial and backyard flocks. ND outbreaks can have substantial economic consequences due to high mortality rates and the imposition of trade restrictions. APMV-1 nucleic acid can be detected from swabs or tissues of suspected cases by PCR. Evidence of infection or vaccination may be demonstrated by the presence of specific antibodies against HN in serum samples. No anti-viral treatments exist, but vaccines are available, although there are currently concerns over their efficacy.

Associations of autoantibodies and clinical profile of the patients with systemic sclerosis

AbstractBackgroundSystemic sclerosis is characterized by the involvement of organs and the presence of specific antibodies. The objectives of this study were to identify the autoantibodies and to determine their association with the selected clinical features of the disease among Bangladeshi systemic sclerosis patients.MethodsThis cross‐sectional study was performed at the rheumatology outpatient clinic of Bangabandhu Sheikh Mujib Medical University. Autoantibodies against nine systemic sclerosis‐specific antigens were tested using an enzyme‐linked immunoassay immunoblot kit. Several clinical features of patients with positive and negative autoantibody were examined by χ2 or Fisher's exact tests.ResultsA total of 71 patients with systemic sclerosis (66; 93.0% female) were included. Their mean age at disease onset was 33.2 years. Fifty‐seven (80.3%) patients had diffuse cutaneous subtype. Out of nine autoantibodies, four were positive, anti‐topoisomerase‐I (57.7%), anti‐U1 ribonucleic protein (21.1%), anti‐RNA polymerase III (18.3%), and anticentromere antibodies (4.2%). Eleven (15.5%) patients were negative for any antibodies and 11 patients were positive for at least two autoantibodies. Anti‐U3‐RNP, anti‐PM‐Scl, anti‐Ku, and anti‐Th/To auto antibodies were absent in all patients. Anti‐RNA polymerase III was associated with raised pulmonary arterial systolic pressure (PASP) and anti‐U1‐RNP with decreased forced vital capacity (FVC).ConclusionsAnti‐topoisomerase‐I was the commonest autoantibody in patients with systemic sclerosis in Bangladesh. Anti‐RNA polymerase III antibody had significant association with raised PASP and anti‐U1‐RNP with decreased FVC.

SARS-CoV-2 seropositivity and COVID-19 among 5 years-old Amazonian children and their association with poverty and food insecurity.

The epidemiology of childhood SARS-CoV-2 infection and COVID-19-related illness remains little studied in high-transmission tropical settings, partly due to the less severe clinical manifestations typically developed by children and the limited availability of diagnostic tests. To address this knowledge gap, we investigate the prevalence and predictors of SARS-CoV-2 infection (either symptomatic or not) and disease in 5 years-old Amazonian children. We retrospectively estimated SARS-CoV-2 attack rates and the proportion of infections leading to COVID-19-related illness among 660 participants in a population-based birth cohort study in the Juruá Valley, Amazonian Brazil. Children were physically examined, tested for SARS-CoV-2 IgG and IgM antibodies, and had a comprehensive health questionnaire administered during a follow-up visit at the age of 5 years carried out in January or June-July 2021. We found serological evidence of past SARS-CoV-2 infection in 297 (45.0%; 95% confidence interval [CI], 41.2-48.9%) of 660 cohort participants, but only 15 (5.1%; 95% CI, 2.9-8.2%) seropositive children had a prior medical diagnosis of COVID-19 reported by their mothers or guardians. The period prevalence of clinically apparent COVID-19, defined as the presence of specific antibodies plus one or more clinical symptoms suggestive of COVID-19 (cough, shortness of breath, and loss of taste or smell) reported by their mothers or guardians since the pandemic onset, was estimated at 7.3% (95% CI, 5.4-9.5%). Importantly, children from the poorest households and those with less educated mothers were significantly more likely to be seropositive, after controlling for potential confounders by mixed-effects multiple Poisson regression analysis. Likewise, the period prevalence of COVID-19 was 1.8-fold (95%, CI 1.2-2.6-fold) higher among cohort participants exposed to food insecurity and 3.0-fold (95% CI, 2.8-3.5-fold) higher among those born to non-White mothers. Finally, children exposed to household and family contacts who had COVID-19 were at an increased risk of being SARS-CoV-2 seropositive and-even more markedly-of having had clinically apparent COVID-19 by the age of 5 years. Childhood SARS-CoV-2 infection and COVID-19-associated illness are substantially underdiagnosed and underreported in the Amazon. Children in the most socioeconomically vulnerable households are disproportionately affected by SARS-CoV-2 infection and disease.

Individual antimicrobial treatments in veal calves: Effect on the net carcasses weight at the slaughterhouse and relationship with the serostatus of the calves upon arrival to the fattening unit

In recent years, the scientific community, institutions, and public opinion have shown a growing awareness towards the problem of antimicrobial resistance. Guidelines from the European Union aim at significantly reducing (even avoiding) the antimicrobial use for prophylactic and metaphylactic purposes in veterinary medicine and this represents an important issue for several intensive farming systems, such as the veal industry. This retrospective observational cohort study analyzed net carcass weight data at the slaughter of 618 veal calves belonging to 6 batches of animals fattened in an Italian commercial farm according to the number of individual antimicrobial treatments administered during the fattening cycle. Sixty-eight percent (419 animals) of the overall sample of calves received at least one individual antimicrobial treatment due to a disease event during the fattening, and 280 of these (45 % of the overall sample) had more than one treatment. On average, the net carcass weight of calves individually treated more than once was about 10 kg lower than that of untreated calves or treated only once. Moreover, the carcasses of calves that received more than one treatment were 16 times more at risk of being severely penalized in payment at the slaughterhouse due to a net weight below 110 kg than those of untreated calves. Serum analysis data of a subsample of 105 calves belonging to 3 out of the 6 batches collected within 5 days after their arrival to the veal farm identified 3 persistently infected animals by bovine viral diarrhea (BVD) virus and a prevalence of calves with serum immunoglobulin concentration below 7.5 g/L of 37 %. The presence of specific antibodies against infectious bovine rhinotracheitis (IBR) and bovine respiratory syncytial virus (BRSV) and a serum immunoglobulin level above 7.5 g/L at the onset of the fattening were the most important factors associated with a decreased risk of multiple individual antimicrobial treatments (and thus, disease development) or with a delay in disease onset during the fattening, thus leading to better performances at the slaughter. The outcomes of this study suggested that a relationship could exist between calf serostatus upon arrival to the veal farm and the possibility to avoid multiple individual antimicrobial treatments during fattening. Further research is needed to deeply study this association, as such knowledge would allow for planning effective preventing strategies to reduce the antimicrobial use in veal calves.

Prevalence of Mycoplasma bovis in Algeria and Characterisation of the Isolated Clones.

Bovine respiratory disease (BRD) is common in calves in Algeria, but to date, Mycoplasma bovis has never been monitored as a potential etiological agent. Here, to assess the presence (direct detection) and circulation (indirect detection) of M. bovis, broncho-alveolar lavage fluids (BALF) and serum samples were collected from 60 veal calf farms in Algeria. A commercial ELISA kit (ID Screen® ELISA) was used to screen for the presence of specific antibodies against M. bovis in 351 blood sera collected from both diseased and healthy calves, and 69% (241 sera) tested positive. BALFs from the 176 diseased calves were used to screen for M. bovis by real-time-PCR (rt-PCR), and 102 (58%) tested positive. A non-exhaustive set of 53 clones were isolated from 44 calves and further subtyped using polC gene sequencing. No predominant subtype was found, and two clones exhibited a new subtype. Fourteen clones were further characterized by multilocus sequence typing, and results showed a high degree of genetic diversity, with some clones having new alleles and subtypes. The minimum inhibitory concentrations (MICs) of 5 antimicrobials regularly used to treat BRD was determined on 45 clones. Susceptibility profiles showed very broad diversity, confirming the variety of clones actively circulating. We detected clones with high MICs, including increased MICs of enrofloxacin (n = 5). This is the first study to report the presence of M. bovis in Algeria in calves with BRD. This research also finds broad genetic and phenotypic diversity in the actively circulating isolates.

Alzheimer's disease diagnosis based on detection of autoantibodies against Aβ using Aβ40 peptide in liposomes

BackgroundAlzheimer’s disease (AD) is the most common form of dementia and affect more than 50 million people worldwide. Thus, there is a high demand by non-invasive methods for an early diagnosis. This work explores the AD diagnostic using the amyloid beta 1–40 (Aβ40) peptide encapsulated into dipalmitoyl phosphatidyl glycerol (DPPG) liposomes and immobilized on polyethylene imine previously deposited on screen-printed carbon electrodes to detect autoantibodies against Aβ40, a potential biomarker found in plasma samples. MethodsThe immunosensor assembly was accompanied by atomic force microscopy (AFM) images that showed globular aggregates from 20 to 200 nm corresponding liposomes and by cyclic voltammetry (CV) through increase of the voltammogram area each material deposited. After building the immunosensor, when it was exposed to antibody anti-Aβ40, there was an increase in film roughness of approximately 9 nm, indicating the formation of the immunocomplex. ResultsIn the detection by CV, the presence of specific antibody, in the range of 0.1 to 10 μg/ml, resulted in an increase in the voltammograms area and current in 0.45 V reaching 3.2 µA.V and 5.7 μA, respectively, in comparison with the control system, which remained almost unchanged from 0.1 μg/ml. In patient samples, both cerebrospinal fluid (CSF) and plasma, was possible separated among positive and negative samples for AD using CV profile and area, with a difference of 0.1 μA.V from the upper error bar of healthy samples for CSF sample and 0.6 μA.V for plasma sample. ConclusionsThese results showed the feasibility of the method employed for the non-invasive diagnostic of Alzheimer’s disease detecting natural autoantibodies that circulate in plasma through a simple and easy-to-interpret method.