Determination Of Immunoglobulin Research Articles

Clinical and epidemiological aspects of primary immunodeficiencies in the Republic of Karakalpakstan

Studying the prevalence of primary immunodeficiency disorders (PID) is an important aspect of epidemiological research, as it allows for assessing the frequency of these diseases occurring in different regions. The distribution of PID may vary depending on ethnic background and geographical location. Information on the prevalence of PID at regional, national, and international levels enables a more accurate determination of the scale of the problem and the development of effective prevention and treatment strategies. Studying the epidemiological and clinical profile of congenital immune disorders (PID) in the Republic of Karakalpakstan is crucial for understanding the prevalence, age and gender structure, as well as the spectrum of nosological forms of these rare diseases. The aim of this study was to investigate the epidemiological and clinical characteristics of congenital immune disorders, primary immunodeficiencies in the Republic of Karakalpakstan. The study material consisted of data on patients receiving inpatient treatment at a multidisciplinary children’s hospital in the Republic of Karakalpakstan and those under the care of an allergist-immunologist in the outpatient clinic at district medical centers in the Republic of Karakalpakstan. The average delay in diagnosis from the onset of clinical manifestation of PID in the region was 2.7 years. It was found that the mortality rate was 2.8%. Screening tests for verifying PID in the diagnostic search stage included clinical and biochemical blood analyses, and determination of serum immunoglobulins. At the time of diagnosis, patients exhibited a significant decrease in IgG and IgA immunoglobulin levels. In addition to the disorder in the humoral branch of adaptive immunity, children with this disease showed a decrease in the absolute number of T lymphocytes. The data obtained from the study indicate insufficient diagnosis of primary immunodeficiencies in the Republic of Karakalpakstan, which may serve as a basis for the development of educational programs aimed at increasing awareness of primary immunodeficiencies. Further research and systematization of information on patients with primary immunodeficiencies are necessary for the development of regional programs for the implementation of screening diagnostics.

Genetic and environmental determinants of Immunoglobulin G in kid serum and adult colostrum of dairy goats

Immune competence is a key factor in the health and resilience of dairy ruminants. The aim of this study was to study immunoglobulin G concentration in serum shortly after birth and in colostrum of lactating animals as proxy of passive immune transfer and colostrum quality, respectively, and thus provide new health and resilience indicators for selection. Data from 424 female kids and 203 primiparous goats, reared on an INRAE experimental farm, were analyzed. IgG concentration was measured in kid serum and goat colostrum, respectively, using the radial immunodiffusion technique. Linear models were used to identify factors of variation in serum and colostrum IgG concentration, and health events were monitored for 5 weeks after birth to measure the impact of passive immune transfer on kids' health. We found a significant effect of serum IgG level on hindquarters cleanliness score (as a proxy of digestive troubles) and nasal and ocular discharge. As expected, the absorption of IgG in the days following birth is essential to ensure the passive transfer of immunity from the mother to the youngster, and provide immune protection. Genetic parameters were estimated for serum IgG concentration in kids and for colostrum characteristics in primiparous goats. Heritability of serum IgG in kids was not significantly different from zero. This low value, with large SE, may be due to the limited sample size and large environmental variability likely not accounted for. Especially, we could not adjust for the amount of IgG that was ingested by the kid, which can have a significant effect on serum IgG and possibly on heritability estimates. Heritability of colostrum traits were moderate, with estimates of 0.25 (±0.16), 0.35 (±0.16), and 0.25 (±0.16) for IgG concentration, IgG mass and colostrum volume, respectively. Genetic correlations with milk production showed high standard errors due to small numbers. However, estimates suggest a negative genetic association between production and IgG concentration in colostrum (rg = -0.44 ± 0.33), probably linked to a positive association with total colostrum volume. The significant heritabilities that were estimated for the first time demonstrate the existence of genetic control of colostrum quality in goats, and open up opportunities for selection in this species.

Tri-signal CdS@SiO2 nanoprobes for accurate and sensitive detection of human immunoglobulin G with enhanced flexibility and internal validation

Accurate and sensitive determination of human immunoglobulin G (HIgG) level is critical for diagnosis and treatment of various diseases, including rheumatoid arthritis, humoral immunodeficiencies, and infectious disease. In this study, versatile tri-signal probes were developed by preparing CdS@SiO2 nanorods that integrate photoluminescence (PL), multi-phonon resonant Raman scattering (MRRS) and infrared absorption (IRA) properties. Through the coating of multiple CdS nanoparticles as cores within SiO2 shells, the PL and MRRS properties of CdS were improved, resulting in a significantly lowered limit of detection (LOD), with the lowest LOD of 12.37 ag mL−1. Integration with the distinctive IRA property of SiO2 shells widened the detection range towards higher concentrations, establishing a final linear range of 50 ag mL−1 to 10 μg mL−1. The remarkable consistency among the three signals highlighted the robust internal verification capability for accurate detection. This approach enhances flexibility in selecting detection methodologies to suit diverse scenarios, facilitating HIgG detection. The tri-signal nanoprobes also exhibited excellent detection selectivity, specificity and repeatability. This study presents a fresh idea for developing high-performance detection strategies.

Nijmegen syndrome in childhood: a clinical case

Nijmegen syndrome is a rare monogenic pathology with an autosomal recessive type of inheritance. The disease is manifested by congenital developmental anomalies and microcephaly, primary immunodeficiency, frequent recurrent viral and bacterial infections, retardation in physical and neuropsychic development. In the medical literature, 150 cases of the syndrome are described; pathology occurs more often among the Slavic population. Nijmegen syndrome belongs to a group of diseases with chromosomal instability. The pathogenetic feature of the syndrome is congenital immunodeficiency of the humoral (B-lymphocytes) and cellular (T-lymphocytes) components. According to statistics, 40 % of children with Nijmegen syndrome are diagnosed with malignant neoplasms. lymphoid tissue is more often affected (non-Hodgkin’s B and T-cell lymphomas, acute lymphoblastic leukemia), and the development of solid neoplasia is also possible. To diagnose Nijmegen syndrome, in addition to assessing the patient clinical status, it is necessary to conduct an extended immunological examination with the determination of immunoglobulins A, M, G and molecular genetic studies.The article presents a clinical case of diagnosis and treatment of Nijmegen syndrome in childhood.

Hantavirus infection in a child: clinical case

Hantavirus infection is a rare zoonotic viral disease characterized by fever, interstitial pneumonia, acute respiratory failure, respiratory distress syndrome, acute heart failure, renal failure and digestive tract disorders. The aim of the work is to acquaint practitioners with the case of hantavirus infection in a child. Materials and methods. The child was treated at Zaporizhzhia City Pediatric Hospital No. 5. He underwent clinical and biochemical blood tests, microbiological, radiological and ultrasound examinations using hospital equipment. Determination of immunoglobulins M and G to hantavirus by immunofluorescence analysis was conducted in a reference laboratory. Results. The child was admitted with complaints of headache, fever, epigastric pain, vomiting and stools with blood, shortness of breath, cough, blood in sputum, muscle pain, decreased diuresis. Laboratory findings included leukocytosis, anemia, thrombocytopenia, elevated ESR, elevated procalcitonin and creatinine levels, and hypercoagulability. Instrumentally, right-sided interstitial pneumonia and duodenal ulcer were diagnosed. The positive result for hantavirus infection was confirmed by the levels of immunoglobulins M and G to hantavirus and the increase in their titers in the dynamics. Comprehensive intensive care resulted in relatively rapid recovery of the child. Conclusions. The criteria for the diagnosis of hantavirus infection with pulmonary and renal syndromes are: the presence of interstitial pulmonary edema, fever with myalgias, arthralgias, cephalgia, nausea, vomiting and headache, followed by oliguria. Neutrophilic leukocytosis, anemia, thrombocytopenia, increased hematocrit, proteinuria, and microhematuria are characteristic. Determination of IgM and G to hantavirus and their dynamics is effective for confirming the diagnosis. Hantavirus infection is rather rare in Ukraine. Clinical discussions of diseases that are rarely occurring in practice are useful for physicians and will facilitate a faster diagnostic search.

Determinants of immunoglobulin G responses to respiratory syncytial virus and rhinovirus in children and adults.

Exposure to respiratory viruses is a significant cause of morbidity and affects virus-specific antibody levels. Little is known about determinants associated with immune response to these viruses. We aimed to investigate the determinants of respiratory syncytial virus (RSV)- and rhinovirus (RV)- specific IgG responses in both children and adults. The study is based on the EGEA cohort, composed of 530 samples of children in EGEA1 (1991-95) and 1241 samples of adults in EGEA2 (2003-07). Cumulative RV-specific IgG levels (species A, B and C) and IgG levels to RSV-G protein were measured by using micro-array technoloy. Multiple linear mixed models (random effect to account for familial dependence) were performed to assess associations between age, sex, body mass index (BMI), tobacco smoke exposure and season of blood sampling with RSV-and RV-specific IgG levels. In children (11.1 ± 2.8 years old, 57% boys), higher RV-specific IgG levels were associated with older age (only for RV-B), female sex and lower BMI, while only older age was associated with higher RSV-specific IgG levels. In adults (43.5 ± 16.7 years old, 48% men), younger age, female sex, lower BMI, active smoking and all seasons except summer were associated with higher RV-specific IgG levels. Older age, active smoking and all seasons except summer were associated with higher RSV-specific IgG levels. Personal and seasonal determinants of RSV- and RV-specific IgG levels seem to vary according to the respiratory virus type and between children and adults, suggesting different patterns of responses along the life course.

Dual-grating waveguide coupler biosensor for the real-time determination of human immunoglobulin M (IgM) antibodies

A dual-grating waveguide coupler biosensor was developed to detect human immunoglobulin M (IgM) antibodies with modification procedures using (3-aminopropyl)triethoxysilane (APTES) as the self-assembled monolayer were optimized. The average bulk sensitivity and optimal resolution were 43.84 ± 1.15 RIU−1 and 6.18 × 10−6 RIU, respectively. Due to the photocatalytic property of TiO2, ultraviolet treatment for 7.5 min was used to form the hydroxyl groups on TiO2 surface. APTES immobilized the biomolecules via chemical bonding; the optimal APTES modified parameters were 2% (v/v) for 0.5 h at 80 °C. Glutaraldehyde (GA) immobilized the protein A without activating carboxyl groups of protein A. The optimal GA modified conditions were 2.5% (v/v) for 48 min. Protein A immobilized the capture antibody directionally, the fragment antigen binding (Fab) region of the capture antibody was more completely exposed to improve the detection effect. Using the optimal modification conditions, the limit of detection for human IgM immunoassay was 2.67 × 10−8 ± 1.18 × 10−8 g/mL. Furthermore, this biosensor was demonstrated to determine the concentration of human IgM within 1 h and has potential for commercialization.

Dietary intervention with edible film-coated multistrain probiotic Lacticaseibacilli in nondairy food matrices significantly increased the recovery of fecal viable Lacticaseibacilli and improved the performance of several colonic biomarkers among slightly malnourished preschool children.

Probiotic enriched dairy products are widely consumed in Western countries for their beneficial effects on the gastrointestinal tract and overall health. The present study aims to investigate the beneficial effects of probiotic Lacticaseibacilli (LAB) strains in non-dairy food matrices. A blend of edible film-coated probiotic LAB, L. plantarum, L. paracasei, and L. rhamnosus, were incorporated into plain biscuits and dry dates. Design of the randomized controlled study: Children of both sexes (mean age 55.7 ± 14.5 months) attending kindergarten in Tersa, a poor urban Giza district, were recruited and randomized into 5 groups of equal numbers. Treatment groups: (1) placebo biscuits, (2) functional probiotic biscuits (0.18 billion colony forming units (cfu) of LAB) (3) functional probiotic + inulin biscuits (0.2 billion cfu of LAB + 2 g of chicory inulin); (4) placebo dates and (5) functional probiotic dates (0.3 billion cfu of viable multistrain LAB). The supplements were served 5 days a week and each child had to consume 21 servings of the supplement. The primary outcome was an increase in the fecal recovery of viable LAB after the intake of 21 servings (T1) compared to the respective baseline counts (T0). The secondary outcomes include the determination of fecal short-chain fatty acids (SCFA) and secretory immunoglobulin A (s-Ig A) using ELISA and fecal ammonia excretion. Results: Statistically significant % increases in the recoveries of fecal viable LAB were found among the children consuming 21 servings of supplements 2, 3, and 5 compared to the respective count at T0. Similar significant increases were found in the fecal concentrations of SCFA and s-Ig A among the children consuming 21 servings of supplements 2, 3 and 5 compared to the respective counts at T0. On the other hand, the concentration of toxic ammonia excretion decreased significantly in the feces of all children consuming probiotic-containing supplements (groups 2, 3, and 5) at T1 compared to the respective concentrations obtained at T0. Conclusion: Multistrain microencapsulated probiotic Lacticaseibacilli in functional biscuits and dry dates successfully tolerated the acidic gastric transit and exerted their bioactive action on the colonic microbiome. The synbiotic supplement exhibited a higher production rate of colonic SCFA. Probiotic-enriched products that confer definitive health benefits are convenient and do not need to be kept under refrigeration. Manipulating the composition and function of the microbiome in childhood through probiotic/+ prebiotic interventions is cost-effective with long-term beneficial health outcomes. This study was approved by the Medical Research Ethics Committee, National Research Center and registered as Clinical Trial 16/422. Written informed consent was obtained from the mothers of all participating children.

Comprehensive insight into the alterations in the gut microbiome and the intestinal barrier as a consequence of iron deficiency anaemia

BackgroundIron deficiency is the top leading cause of anaemia, whose treatment has been shown to deteriorate gut health. However, a comprehensive analysis of the intestinal barrier and the gut microbiome during IDA have not been performed to date. This study aims to delve further into the analysis of these two aspects, which will mean a step forward minimising the negative impact of iron supplements on intestinal health. MethodsIDA was experimentally induced in an animal model. Shotgun sequencing was used to analyse the gut microbiome in the colonic region, while the intestinal barrier was studied through histological analyses, mRNA sequencing (RNA-Seq), qPCR and immunofluorescence. Determinations of lipopolysaccharide (LPS) and bacteria-specific immunoglobulins were performed to assess microbial translocation. ResultsMicrobial metabolism in the colon shifted towards an increased production of certain amino acids, short chain fatty acids and nucleotides, with Clostridium species being enriched during IDA. Structural alterations of the colonic epithelium were shown by histological analysis. RNA-Seq revealed a downregulation of extracellular matrix-associated genes and proteins and an overall underdeveloped epithelium. Increased levels of serum LPS and an increased immune response against dysbiotic bacteria support an impairment in the integrity of the gut barrier during IDA. ConclusionsIDA negatively impacts the gut microbiome and the intestinal barrier, triggering an increased microbial translocation. This study emphasizes the deterioration of gut health during IDA and the fact that it should be addressed when treating the disease.

Оцінка сироваткового загального та специфічного імуноглобуліну Е в дітей з персистуючим алергічним ринітом і поєднаним із бронхіальною астмою при сенсибілізації до алергенів кліщів

Allergic rhinitis (AR) is a common immunoglobulin E (IgE) dependent disease among allergic patients worldwide. At the same time, the determination of specific IgE in the blood serum allows characterizing the relevant sensitizing allergens. Despite the existence of conditional thresholds for normal and elevated levels of total and specific IgE, there is no consensus in the literature on which IgE value unequivocally confirms the presence of atopy and what is the relationship between these levels and the severity of clinical manifestations. Purpose - to evaluate the levels of serum total and specific IgE and their relationship with severity and age in children with persistent AR and combined with bronchial asthma (BA) during sensitization to house dust mite (HDM) allergens. Materials and methods. A clinical, anamnestic and laboratory examination of 259 children aged 5 to 17 years with AR and combined AR with BA, who were sensitized to HDM allergens, was conducted. Results. In AR in children with sensitization to HDM, there is a dependence of serum total IgE level on the severity of AR and age of children. With increasing severity of AR manifestations, a significant (p<0.05) increase in total IgE levels was noted. It was found that the content of sIgE also increased with increasing severity of AR. Thus, statistically significantly (p<0.05) higher levels of sIgE were determined in sensitization to the major HDM molecules Der f 1, Der p 1, Der p 2, Der p 23 in children with severe AR. Sensitization to the tropomyosin Der p10 molecule, which usually has no effect on the clinical manifestations of respiratory allergy but indicates cross-reactivity to seafood, was significantly more common (p<0.05) in children aged 3-7 years compared to children aged 13-17 years. Conclusions. The data obtained partially define the pathogenetic features in the formation of sensitization to HDM and their individual molecules, confirming that this process begins at an early age, which may indicate the need to start allergen-specific immunotherapy in such children if clinically necessary. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

Determination of minimum individual cow colostral immunoglobulin G concentration required to provide adequate total immunoglobulin G mass in pooled colostrum fed to Jersey calves.

Quantify the minimum individual cow colostral immunoglobulin G (IgG) concentration required for pooling to achieve adequate transfer of passive immunity in calves. 201 Jersey cows. Colostrum was collected from 28 pools and heat treated before being fed to calves or stored. Parity, total number of cows contributing to the pool, individual cow colostral volume contributions, and total volume of each colostrum pool were recorded. Colostrum IgG concentrations in individual and pooled (pre- and post-heat treatment) samples were analyzed by radial immunodiffusion and Brix refractometry. Colostral IgG concentration of ≥ 50g/L was considered the current recommended dairy industry standard for acceptable colostrum quality. Multivariable models were performed to determine factors affecting pooled colostral IgG concentrations. The minimum colostral IgG concentration required for pooling to achieve the recommended total mass of at least 200g IgG to be fed to a calf was calculated. Total pool volume and the number of cows contributing to the pool were significant factors affecting IgG concentration. Colostrum pools from ≤ 7 cows, with a minimum pool IgG concentration of 70.4 g/L (22.9% Brix) or colostrum pool volume ≤ 40 L, with a minimum pool IgG concentration of 66.2 g/L (21.8% Brix) achieved the recommended total mass of at least 200g IgG in 4L of colostrum. When feeding pooled colostrum, IgG concentrations higher than the industry standard of 50 g/L is recommended to reduce the risk of failure of transfer of passive immunity in calves.

A systematic approach to assessing the effectiveness of personal protective equipment for the skin from the effects of industrial factors

Introduction. The causes of occupational skin diseases in 40% of cases are in the absence or unsatisfactory condition of dermatological personal protective equipment. Professional protective skin care remains the main requirement of occupational safety.
 The concept of skin screening is giving way to a new approach — the restoration of its own epidermal barrier. Etiopathogenetic prevention technologies are based on inactivation of irritating chemicals and allergens, binding and neutralization of heavy and alkaline earth metal ions, as well as platinoids. The safety assessment of dermatological personal protective equipment is regulated by the state and in accordance with TR CU 019/2011 "On the safety of personal protective equipment", and the effectiveness assessment is voluntary for manufacturers.
 The study aims to develop a scheme for conducting clinical testing of evaluation of dermatological personal protective equipment for workers exposed to certain production factors.
 Materials and methods. Skin tests are a standard method of allergological diagnosis.
 In order to exclude additional sensitization that occurs when taking skin samples with a production allergen, for the reliability of detecting preclinical manifestations of sensitization, the researchers used an in vitro diagnostic method (determination of specific immunoglobulin E — Ig E). To assess the effectiveness of dermatological personal protective equipment on persons working in contact with nickel-containing compounds, the authors determined the nickel content in the blood by atomic absorption method. They evaluated the barrier function of the skin using the "Skin-o-mat" device from "Cosmomed GmbH", Germany.
 Results. The specialists proposed a scheme for conducting clinical testing of the evaluation of dermatological personal protective equipment for workers, including an assessment of the barrier function of the skin and in vitro diagnostic methods. The authors evaluated the parameters in dynamics — before and after using. This scheme makes it possible to exclude additional sensitization of employees.
 Limitations. The study is limited by the number of workers who underwent periodic medical examinations at the clinic of Izmerov Research Institute of Occupational Health who have contact with nickel compounds in the workplace.
 Conclusion. The etiopathogenetic clinical approbation of the evaluation of the effectiveness of dermatological personal protective equipment on workers in production conditions includes an assessment of the skin and an assessment of the barrier-protective function of the skin with the fixation of indicators. To study the effectiveness of skin protection products, an important point is the rational and reasonable selection of groups of approbants on which the study of the protective properties of these products will be carried out.
 Ethics. The authors have carried out the study in accordance with the Ethical standard set out in the Helsinki Declaration of the World Medical Association of 1964 (with amendments and additions from 2013).

Molecular detection and association of HIV and HSV-2 IgM co-infection among asymptomatic pregnant women in Port Harcourt, Nigeria

HSV-2 is a common human pathogen that can cause primary and recurrent infection of the mucous membrane. Genital infection with HSV-2 facilitates the acquisition of HIV, both mutually reinforcing infection. Infection with herpes simplex virus type 2 is a significant public health problem being the leading cause of genital ulcerative disease. This study aimed at Molecular investigation and association of HIV and HSV-2 IgM co-infection among asymptomatic pregnant women in Port Harcourt, Nigeria. This is a cross-sectional study conducted at the antenatal unit of the University of Port Harcourt Teaching Hospital (UPTH), River State. A total of 168 consenting pregnant women were selected. The study involved collecting socio-demographic data and laboratory determination of HSV-2 immunoglobulin M (IgM) and HIV seroprevalence using enzyme linked immunosorbent assay (DIAPRO Diagnostic Bioprobes, Milan, Italy) and ALERE determine/STAT PAK respectively. Data analysis was done using graph pad statistical package. Chi square analysis was used to determine the correlation of the infection with socio-demographic factors. The seroprevalence for HSV-2 IgM was 29.8% (50/168) of the respondents while there was no co-infection between IgM and HIV. All HIV respondents were married. Majority are trader having tertiary educational qualification and are in a monogamous relationship. With regard to IgM seropositivity age, marital status, religion and gestation period were significantly associated with IgM while educational level, occupation, type of family and parity were not significantly associated with HSV-2 IgM. There is a significant correlation between HIV seropositivity with parity and history of sexually transmitted disease while there is no significant difference between HIV prevalence with age, marital status education level occupation type of family, religion history of abortion and the prevalence of HSV-2 was high among women of childbearing age in Port Harcourt, Nigeria. Effort should be made to increase the awareness of HSV-2 and HIV infection among the childbearing populace and serological testing of pregnant women should be included as a routine test.

A new approach to immunosuppressive therapy in patients with glomerulonephritis with nephrotic syndrome

Glomerulonephritis (GN) is a group of immuno-inflammatory kidney diseases with predominant glomerular lesions that are difficult to treat. The greatest problems are caused by the treatment of GN with nephrotic syndrome, which often has a recurrent course. The aim of the research was to study the effectiveness of recombinant interleukin-2 (rIL-2) therapy in the GN patients with nephrotic syndrome. 62 patients with a nephrotic form of primary GN with frequent relapses admitted to the Nephrology Department have been recruited into the study. The age of patients was from 18 to 65 years. The patients underwent standard examinations, as well as immunological studies, before administration of the anti-relapse treatment, and 12 months after the treatment was started. Immunological testing included immunophenotyping of lymphocytes with counting of T and B lymphocytes, immunoregulatory and activated subpopulations of T lymphocytes, determination of urinary immunoglobulins (IgM, IgG, IgA) by immunoturbidimetric assays, proinflammatory cytokines – IL-1β, IL-8, IL-17A and anti-inflammatory cytokine IL-10 by ELISA tests. As a result of studies, the examined patients showed an increased contents of T helper cells, activated T lymphocytes (CD8+HLA-DR+CD45+, CD3+CD8brightCD38+) along with decreased numbers of Treg cells and an increased contents of proinflammatory cytokines IL-1β, IL-8, IL-17A and immunoglobulins of all three classes in urinary samples.The cohort of patients with GN selected for the study was divided in two groups (the main group and the comparison group). In addition to nephroprotective and steroid therapy, the treatment regimen of patients included rIL-2 in the main group, or cyclophosphamide in the comparison group. Regardless of the method used, the levels of protein, IgG and IL-17A in the urine proved to be decreased relative to the initial values; the contents of B cells and HLA-DR+ cytotoxic T lymphocytes in peripheral blood were found to be decreased. The revealed changes were more pronounced in the main group of patients. By 12 months after starting the treatment, the mentioned indexes began to differ significantly in the main group from those in the comparison group. Serum creatinine levels, numbers of T helper cells and Treg cells, IL-1β levels in urine did not undergo significant changes in the comparison group, whereas a decrease in serum creatinine and urinary IL-1β was registered in the main group of patients, along with decreased number of T helpers and increased numbers of Treg cells. In the main group of patients treated with rIL-2, the average number of relapses per year decreased by 4 times, showing only a 1.2-fold decrease in the comparison group. Hence, the low-dose therapy with rIL-2 may be considered an effective and safe alternative to conventional immunosuppressive therapy and a new option of the targeted treatment of glomerulonephritis with frequent recurrence of nephrotic syndrome.

Fenofibrate attenuates asthma features in an ovalbumin-induced mouse model via suppressing NF-κB binding activity

Background/aimAsthma is a chronic inflammatory disease of the airways with a high prevalence. Asthma has a complex pathophysiology and about 5–10% of patients are not fully responsive to the currently available treatments. The aim of this study is to investigate the involvement of NF-κB in the effects of fenofibrate on a mouse model of allergic asthma. Materials and methodsA total of 49 BALB/c mice were randomly distributed into 7 groups (n = 7). Allergic asthma model was created by administering i.p. injections of ovalbumin on days 0, 14 and 21, followed by provocation with inhaled ovalbumin on days 28, 29 and 30. Fenofibrate was orally given in 3 different doses; 1, 10 and 30 mg/kg through days 21–30 of the experiment. On day 31, pulmonary function test using whole body plethysmography was performed. The mice were sacrificed 24 h later. Blood samples were obtained, and serum of each sample was separated for IgE determination. Bronchoalveolar lavage fluid (BALF) and lung tissues were collected to measure IL-5 and IL-13 levels. Nuclear extracts of lung tissues were employed to assess nuclear factor kappa B (NF-κB) p65 binding activity. ResultsEnhanced Pause (Penh) values were significantly increased in ovalbumin-sensitized and challenged mice (p < 0.01). Administration of fenofibrate (10 and 30 mg/kg) resulted in improved pulmonary function as shown by significantly lower Penh values (p < 0.01). Interleukin (IL) − 5 and IL-13 levels in BALF and lung tissues and immunoglobulin E (IgE) levels in serum were significantly elevated in the allergic mice. IL-5 levels in the lung tissues of mice treated with 1 mg/kg fenofibrate (FEN1) group were significantly reduced (p < 0.01). BALF and lung tissue IL-5 and IL-13 levels in mice treated with 10 and 30 mg/kg fenofibrate, FEN10 and FEN30, respectively, were significantly diminished when compared to the ovalbumin-treated (OVA) group, whereas treatment with 1 mg/kg fenofibrate resulted in insignificant changes. IgE levels in the serum of FEN30 group mice have shown a prominent reduction (p < 0.01). NF-κB p65 binding activity was higher in mice sensitized and challenged with ovalbumin (p < 0.01). NF-κB p65 binding activity was significantly reduced in allergic mice treated with 30 mg/kg (p < 0.01) fenofibrate. ConclusionsIn this study, we showed that administration of 10 and 30 mg/kg fenofibrate effectively attenuated airway hyperresponsiveness and inflammation in a mouse model of allergic asthma, possibly through inhibition of NF-κB binding activity.

Immunoassay based on peroxidase silver conjugate for the determination of human immunoglobulins against tick‐borne borreliosis (lyme disease) by voltammetry and spectrophotometry

AbstractIn this work two strategies for the synthesis of peroxidase silver conjugates for the qualitative and quantitative determination of immunoglobulins (IgG) to ixodid tick‐borne borreliosis (ITBB) (Lyme disease) in human serum were proposed. The first approach for Ab‐HRP@AgNP conjugate synthesis involved silver nanoparticles (Ag NPs) capped with a commercial peroxidase conjugate (Ab‐HRP) by passive adsorption. The second strategy was based on the initial coupling of Ag NPs with human anti‐species antibodies (Ab) by passive adsorption followed by the introduction of horseradish peroxidase (HRP) enzyme into the reaction mixture as a blocking reagent for Ab@AgNP@HRP conjugate synthesis. The formation of peroxidase silver conjugates was proved by UV/Vis spectroscopy and Transmission Electron Microscopy (TEM). The catalytic activity of Ab‐HRP@AgNP and Ab@AgNP@HRP conjugates was evaluated by Michaelis‐Menten kinetics. A commercially available 96‐well microtiter plate with recombinant antigens to ITBB was used as a platform for immobilization of analyzed IgG. The HRP in Ab‐HRP@AgNP conjugate was found to retain a sufficient level of activity for interaction with the H2O2 substrate to form an intensely colored reaction product. Therefore Ab‐HRP@AgNP conjugate can be used in enzyme‐linked immunosorbent assay (ELISA) with spectrophotometric detection of 3,3’,5,5’‐Tetramethylbenzidine (TMB Ox) for quantitative determination of IgG to ITBB in human serum in the concentration range 12.5–800 ng ml−1 with LOD 2 ng ml−1. Ab@AgNP@HRP conjugate is recommended for the electrochemical determination of IgG to ITBB in human serum at LOD 3 ng ml−1 with registration of silver oxidation by linear sweep anodic stripping voltammetry (LSASV). Ag NPs in Ab‐HRP@AgNP and Ab@AgNP@HRP conjugates do not change electrochemical activity during storage and can be used as an electrochemical label in LSASV method in case of HRP inactivation. The immunoassay based on peroxidase silver conjugates expands the analytical potential for the determination of IgG to ITBB especially during the period of increasing incidence.

Dietary Supplementation of Brevibacillus laterosporus S62-9 Improves Broiler Growth and Immunity by Regulating Cecal Microbiota and Metabolites.

Brevibacillus laterosporus has been added as a direct-fed microbiota to chicken. Yet, few studies have reported the effects of B. laterosporus on broiler growth and gut microbiota. The aim of this study was to evaluate the effects of B. laterosporus S62-9 on growth performance, immunity, cecal microbiota, and metabolites in broilers. A total of 160 1-day-old broilers were randomly divided into S62-9 and control groups, with or without 106CFU/g B. laterosporus S62-9 supplementation, respectively. During the 42days feeding, body weight and feed intake were recorded weekly. Serum was collected for immunoglobulin determination, and cecal contents were taken for 16S rDNA analysis and metabolome at Day 42. Results indicated that the broilers in S62-9 group showed an increase in body weight of 7.2% and 5.19% improvement in feed conversion ratio compared to the control group. The B. laterosporus S62-9 supplementation promoted the maturation of immune organs and increased the concentration of serum immunoglobulins. Furthermore, the α-diversity of cecal microbiota was improved in the S62-9 group. B. laterosporus S62-9 supplementation increased the relative abundance of beneficial bacteria including Akkermansia, Bifidobacterium, and Lactobacillus, while decreased the relative abundance of pathogens including Klebsiella and Pseudomonas. Untargeted metabolomics revealed that 53 differential metabolites between the two groups. The differential metabolites were enriched in 4 amino acid metabolic pathways, including arginine biosynthesis and glutathione metabolism. In summary, B. laterosporus S62-9 supplementation could improve the growth performance and immunity through the regulation of gut microbiota and metabolome in broilers.

Evolution of laboratory parameters in patients with lipid transfer protein syndrome after 3 years of immunotherapy.

Lipid transfer protein (LTP) syndrome is an increasingly prevailing disease, especially in the young population, with severely affected quality of life. Since 2013, a specific treatment, called sublingual immunotherapy (SLIT), with peach extract (SLIT-peach®) has been used, but with no long-term effectiveness studies. The main objective of the present study was to assess the long-term effectiveness of SLIT-peach® and to relate the clinical evolution of patients. This was an ambispective study conducted for 3 years. A total of 25 patients with LTP syndrome were selected and treated with SLIT-peach®. They underwent a provocation test in the first year with reintroduced foods that had produced symptoms in the past. Analytical determination of specific immunoglobulin E (IgE) and immunoglobulin G4 (IgG4) to peach (Pru p 3) was performed at the beginning of treatment, at the first year of initiation, and at the end of treatment. These data were compared with the control group comprising 14 patients with LTP syndrome without treatment. A statistically significant decrease in specific IgE to Pru p 3 at the end of the treatment and an increase in specific IgG4 to Pru p 3 1 year after treatment initiation were observed in the active group in relation to tolerance to foods with LTPs. These results indicate that food tolerance begins after the first year and is maintained after the end of 3 years of treatment. In conclusion, treatment with SLIT-peach® for 3 years is effective for patients with LTP syndrome, preventing the evolution of the disease, allowing patients to restart a diet with plant foods, and improving their quality of life.

Clinical laboratory diagnostics of antibodies to SARS-CoV-2: from a QR code to the reality

Background: The immune response to SARS-CoV-2 includes the production of specific immunoglobulins to protein antigens of SARS-CoV-2. Depending on the type and level of immunoglobulins, it is possible to assess the stage of the disease and evaluate the effectiveness of vaccination. The main approach to the determination of immunoglobulins to SARS-CoV-2 in human biological fluids is enzyme-linked sorbent immunoassay. Its data, in particular, are used to issue an electronic COVID-19 certificate with a QR code. However, the qualitative and quantitative composition of immunoglobulins for a QR code is not officially regulated. Aim: measuring the immunoglobulins level in the human blood serum with different types of immunity to the new coronavirus infection (COVID-19) to select the most informative indicators of protective immunity. Methods: The study included 76 blood serum samples from male and female volunteers (age, 18 to 50 y.o.) in compliance with the ethical standards. The detection of IgA, IgM, IgG (total to different regions of SARS-CoV-2, S-protein IgG and RBD-fragment IgG), IgG avidity, and the level of the SARS-CoV-2 N-antigen was performed by enzyme-linked immunosorbent assay (ELISA) using commercially available reagent kits. Results: The indicators of the level of antibodies (both "protective" IgG and IgA of the initial phase of infection) are most pronounced in persons who have been vaccinated and have had COVID-19, and least pronounced in unvaccinated people. For recovered unvaccinated individuals, the level of total protective antibodies and IgG to the S-protein, including the RBD fragment, is the lowest; the avidity of IgG is lower than that in the other groups, too. The IgG avidity in vaccinated patients is higher than that in recovered ones. It should be noted that there were no differences in the level of both total IgG to SARS-CoV-2, to the S-protein and to the RBD-fragment of the S-protein for recovered and vaccinated individuals. Conclusion: The analysis of COVID-19 immunoglobulins indicates a different profile of the humoral immune response following vaccination and previous infection with COVID-19. To quickly assess the immune response to previous and current COVID-19 infection, as well as to detect the post-vaccination immunity, it is advisable to use the total level of IgG to SARS-CoV-2. For deeper assessment of protective immunity and production of protective antibodies, it is better to evaluate the quantitative content of IgG to the S protein and its RBD fragment. The equal level of IgA in the experimental groups indicates an ongoing interaction with SARS CoV-2 in the population. Thus, the electronic COVID-19 certificate is of little use when it is formed by only one of the indicators without taking into account the rest.

Reversed-phase allergen microarrays on optical discs for multiplexed diagnostics of food allergies

A high percentage of the population suffers from multiple food allergies justifying the importance of reliable diagnostic methods. Single-analyte solutions based on the determination of specific immunoglobulins E (sIgE) are safe and fast but are generally time-consuming and expensive. Thus sustainable microanalytical methods that provide multianalyte profiling information are highly demanded. This work presents the in vitro biosensing of specific IgE levels based on a reversed-phase allergen array. The approach consists of optical biosensing supported by direct multiplex immunoassays and on-disc technology. It identifies 12 sIgE associated with food allergies in a single analysis with a low serum sample volume (25 µL). After processing captured images, specific signals for each target biomarker correlate to their concentration. The assay analytically performs well with 0.3 IU/mL and 0.41 IU/mL as the detection and quantification limits in serum, respectively. This novel method achieves excellent clinical specificity (100%) and high sensitivity (91.1%), considering the diagnosis obtained by clinical history and ImmunoCAP analysis. The results demonstrate that microanalytical systems based on allergen arrays can potentially diagnose multiple food allergies and are easily implemented in primary care laboratory settings.Graphical