Immunoglobulin Class Research Articles

IgA: Structure, Function, and Developability.

Immunoglobulin A (IgA) plays a key role in defending mucosal surfaces against attack by infectious microorganisms. Such sites present a major site of susceptibility due to their vast surface area and their constant exposure to ingested and inhaled material. The importance of IgA to effective immune defence is signalled by the fact that more IgA is produced than all the other immunoglobulin classes combined. Indeed, IgA is not just the most prevalent antibody class at mucosal sites, but is also present at significant concentrations in serum. The unique structural features of the IgA heavy chain allow IgA to polymerise, resulting in mainly dimeric forms, along with some higher polymers, in secretions. Both serum IgA, which is principally monomeric, and secretory forms of IgA are capable of neutralising and removing pathogens through a range of mechanisms, including triggering the IgA Fc receptor known as FcαRI or CD89 on phagocytes. The effectiveness of these elimination processes is highlighted by the fact that various pathogens have evolved mechanisms to thwart such IgA-mediated clearance. As the structure–function relationships governing the varied capabilities of this immunoglobulin class come into increasingly clear focus, and means to circumvent any inherent limitations are developed, IgA-based monoclonal antibodies are set to emerge as new and potent options in the therapeutic arena.

PSXIV-5 Results of external use of staphylococcal toxoid for the prevention of mastitis in cows

Abstract Mastitis in cows in the farms of the Russian Federation cause great economic damage. Economic losses include the cost of treating cows and the cost of recycled milk. In the farms of the North-West region of the Russian Federation mastitis occurs in 20–30% of lactating cows. An important task is to find effective ways to prevent mastitis. The task of our research included the study of the effectiveness of external use of staphylococcal toxoid. For the experiment, 2 experimental groups of dry cows of 10 animals each were formed. For the first group of cows, 5 days before the expected date of calving, an ointment containing staphylococcal toxoid and dimethyl sulfoxide was applied to the area of the milk mirror. Udder treatment was performed once a day for five days. Cows of the second group were immunized with Starvac® vaccine in accordance with the instructions, twice during the dry period. The third group of cows (n = 20) was the control. After calving, the concentration of immunoglobulins in colostrum was examined in all cows. In cows of the first group, the concentration of immunoglobulins G, M and A in colostrum was 8.59 ± 0.9; 1.1 ± 0.13 and 5.42 ± 0.9 g / l, respectively. The concentration of immunoglobulins in cows of the first group was higher, on average, by 1.3–2.0 times in comparison with cows that were given the Starvac vaccine. In comparison with the control group, the growth of all classes of immunoglobulins was determined by 2.2 - 5.2 times (P < 0.01). The results obtained showed high efficacy of external use of staphylococcal toxoid together with dimethisulfoxide.

Systemic Immune Profile in Patients With CRSwNP.

The immune pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remains obscure. Our aim was to compare humoral immunity and white blood cell counts in patients with CRSwNP and controls. A prospective case-control study was carried out in 37 patients with CRSwNP and 34 controls without CRS. Clinical data were gathered through a systematic interview. Computed tomography scan, skin prick test, spirometry, and immunological parameters (leukocyte differential count, immunoglobulin classes, and immunoglobulin [Ig] G subclasses) in serum specimens were obtained. Statistical analysis was performed using SPSS v.23. The prevalence of chronic lower respiratory diseases was greater in the CRSwNP group (P < .001), but atopic disease had no significant difference. A significantly higher eosinophil (P < .001) and basophil relative count (P = .022) and a lower relative neutrophil count (P = .013) were found among CRSwNP group. Patients with CRSwNP had higher IgG1 (P = .022), but lower IgG2 (P = .014) and IgG3 (P = .018) serum levels compared to controls; IgG4, total IgG, IgA, IgM, and IgE serum levels did not differ between groups, as well as the prevalence of immunoglobulin classes or IgG subclasses deficiency. The variation observed in peripheral relative leukocyte count and the systemic IgG1 subclass shift are similar to what is known to happen in nasal polyp tissue. A unique systemic immune profile seems to be present in patients with CRSwNP.

Influence of ultra-small particles (10-20 nm) in the atmospheric air of Chernivtsi during intrauterine development on the course of sepsis in newborns

Unfavorable environmental factors that affect parents cause impaired programmed development, the fetus, especially in its sensitive period. Even environmental factors of low intensity due to the bioaccumulation of xenobiotics in maternal tissues and their release to the bloodstream during pregnancy can have a pathogenic effect on the fetus, especially during sensitive periods of its development. To analyze the influence of the content of ultra-small particles (10-20 nm) in the atmospheric air during prenatal development on the course of sepsis in newborns. Based on the results of the analysis of content in the atmospheric air, two clinical observation subgroups were formed. The first subgroup included 17 patients with neonatal sepsis in whom the UFPs content in the air exceeded1.0 inone of the periods. The second subgroup consisted of 35 newborns with sepsis in whom the content of UFPs 10-20 nm in the atmospheric air was less than1.0 inone of these periods. The content in the atmospheric air of ultra-small particles (10-20 nm) during the sensitive period of pre-natal development did not significantly affect the gestational age of newborns, the content in their blood in the presence of sepsis of interleukins-6, -8, -10, pressepsin, procalcitonin, C-RP and immunoglobulins of classes A, G, M, as well as the duration of intensive care and inpatient care.

On significance of immunoglobulin class G to glycoprotein E in diagnosis of varicella-zoster virus

Purpose.To evaluate the significance of IgG detection to glycoprotein E of varicella-zoster virus (VZV) for the infection stage diagnosis.Materials and methods.Integration of literature data.Results.Analysis of literature data allows to make quite reasonable conclusion that the dynamics of IgG antibodies to glycoprotein E of VZV is subject to the general laws of the antibody productions in the course of infectious processes, although the final completion of this discussion the essential results of direct detection of the dynamics of these antibodies in various stages and forms of the indicated infection.

Efficiency of Human Plague Vaccination in Tuvinian Natural Plague Focus

Background. Plague is an especially dangerous natural focal infectious disease belonging to a group of quarantine infections. There are eleven natural plague foci in Russian Federation. In Republic Tyva plague endemic territories include Ovyur, Mongun-Taigin and Tes-Hem areas where Y. pestis strains are intermittently isolated from Citellus undulates. Population living at the territory of the natural foci get immunoprophylaxis against plague at complication of epizootic and epidemic conditions.This paper presents the results of monitoring indicators of the immune status of people vaccinated with the plague vaccine living in the territory of the Tuva natural focus.Materials and methods. The study involved 76 volunteers who had not previously been vaccinated. The study included the determination of production IFN-γ, IL-4, TNF-α by blood cells, titers of specific IgG antibodies to the capsule F1 antigen of the Yersinia pestis, and concentrations of immunoglobulins in serum blood, as well as immunophenotyping of blood lymphocytes.Results. In the course of a comprehensive immunological study, features of the development of cellular and humoral reactions in people living in the territory of the Tuva natural plague focus were established in the first months after vaccination. Changes in the concentration dynamics of the main classes of immunoglobulins were accompanied by an increase in the level of specific IgGs to the F1 within 6 months after immunization. In the same period, a significant increase in the production of cytokines, as well as significant changes in terms of the subpopulation composition of the vaccinated blood.Conclusion. It is necessary to note the importance of studying of the human immune status in 1–3 months after plague vaccination as this period coincides with potentially dangerous season from epidemiological point of view. Nevertheless, much important role for improvement of tactics of the specific prevention measures plays the data received after the revaccination.

Utility of Newborn Screening for Severe Combined Immunodeficiency and X-Linked Agammaglobulinemia Using TREC and KREC Assays

Background Severe combined immune deficiency (SCID) is a potentially fatal primary immunodeficiency due to the absence of T and B lymphocyte function. Early intervention for patients with SCID results in a higher survival rate. From 2017, we launched the first optional newborn screening (NBS) for SCID in Japan based on the detection of T-cell receptor excision circles (TREC). However, NBS for severe B-cell lymphopenia, such as X-linked agammaglobulinemia (XLA), has not been a standard screening test because of a high false-positive rate of Kappa-deleting recombination excision circles (KREC), which reflects the replication of B cells. XLA is characterized by severe B-cell lymphopenia and marked reduction of all classes of serum immunoglobulins. Patients with XLA require early diagnosis and immunoglobulin replacement therapy to prevent the development of bronchiectasis caused by recurrent infections. This study aimed to analyze the results of NBS for SCID and elucidate the utility of NBS for SCID and XLA using the TREC/KREC assay. Patients and Methods We enrolled infants who received NBS for SCID (n = 29,447) between April 2017 and June 2018. Using the EnLiteTM TREC kit, we measured TRECA and β-actin, which are used as controls for monitoring sample amplification. Samples with less than 30 copies/µL and adequate β-actin were defined as positive TRECA. All infants with positive TRECA were followed up for at least 12 months. We measured TRECB and KREC using the EnLiteTM TREC/KREC kit in these infants. As positive controls, we used TRECB and KREC in patients with SCID and XLA, respectively. Furthermore, all infants with positive TRECA were evaluated using flow cytometric analysis and target capture-based next-generation sequencing (NGS) analysis covering 349 primary immunodeficiency- and bone marrow failure-related genes to evaluate CD4+CD45RA+ T-cell counts and identify diagnostic variants. This study was approved by the institutional review board of Nagoya University Graduate School of Medicine. Results Of the infants who underwent NBS for SCID, 43 (0.15%) infants showed positive TRECA. All 43 infants were followed up in Nagoya University for at least 12 months. Of these, we identified one case with DiGeorge syndrome showing severe lymphopenia but did not identify typical SCID. TRECB and KREC were measured in 43 infants with positive TRECA. To determine which kit is more useful to detect T-cell lymphopenia, we compared TRECA with TRECB in 1454 infants with normal TRECA and 43 with positive TRECA. All healthy infants with normal TRECA showed TRECB with more than 30 copies/µL but nine patients with SCID showed extremely low TRECB (median [range], 0 [0-3] copies/µL). Only 6 of 43 (14%) infants showed TRECB with less than 30 copies/µL. Moreover, we analyzed the correlation between CD4+CD45RA+ T-cell counts and TRECB. Compared with 37 infants with normal TRECB, 6 infants with positive TRECB demonstrated significantly lower CD4+CD45RA+ T-cell counts (P = 0.026). However, target capture-based NGS did not identify any diagnostic variants among them. This finding suggested that using this kit, false-positive rates might be decreased from 0.15% (43/29,447) to 0.02% (6/29,447). Using this kit, we assessed KREC in 1454 infants with normal TRECA and 43 with positive TRECA. Of these, we identified one case with less than 30 copies/µL KREC who was diagnosed with congenital asplenia. As positive controls, all six patients with XLA showed quite low KREC (0 [0-9] copies/µL). Compared with previously reported KREC assay, this kit may result in lower false-positive rates. Furthermore, to demonstrate whether KREC reflects the replication of B cells, we analyzed the correlation with CD19+ B-cell counts and KREC. Among 43 infants with positive TRECA, infants with less than 500/µL CD19+ B cells showed significantly lower KREC than those with more than 500/µL CD19+ B cells (P = 0.014). Conclusion We conducted the first large-scale study to evaluate the utility of the newly released EnLiteTM TREC/KREC kit. This kit may be more useful than the current TREC kit to identify infants with T-cell lymphopenia and to avoid unnecessary follow up. Compared with previous NBS for XLA, the false-positive rate of this assay was within an acceptable range. Furthermore, TRECB and KREC were assessed with almost the same screening cost and labor. Therefore, we are considering switching from the current TREC kit to this TREC/KREC kit. Disclosures No relevant conflicts of interest to declare.

CYT-0853, a Novel RAD51 Inhibitor, Modulates Immunoregulatory B-Lymphocytes

Activation Induced Cytidine Deaminase (AICDA/AID) is a DNA-directed cytidine deaminase that is normally only expressed in activated B-cells to promote somatic hypermutations and immunoglobulin class switching. In cancer cells, AID causes significant genotoxic stress through DNA replication fork damage, creating a dependency upon the homologous recombination repair factor, RAD51, for survival. We have demonstrated anti-cancer activity through disruption of this axis in multiple preclinical lymphoid cancer models. Autoreactive B cells depend on RAD51 for survival and are chronically auto-stimulated and therefore continually re-express AID. It has been shown that ectopic expression of AID in autoreactive B-cells causes genome-wide DNA damage (similar to cancers). Given the role of autoreactive B cells and autoantibodies in autoimmune disorders, we hypothesize that immunomodulation of B cells via the RAD51/AID axis will remediate inflammatory disease processes. Our previous data suggests that RAD51 modulation enhances the CD73+ B cell population and reduces antibody diversity in T1D mice, indicating precise effects on AID-mediated antibody diversification. CYT-0853 is a novel RAD51 inhibitor that sensitizes cells to AID activity. Here, we assessed the in vivo effect of CYT-0853 on primary B cells and antibody production. Wild-type C57BL/6 mice were treated with 40mg/kg CYT-0853 or vehicle for five weeks. One-week post-treatment start, mice were immunized with DNP-KLH antigen mixed with Complete Freund's Adjuvant. A second booster with DNP-KLH antigen mixed with Incomplete Freund's Adjuvant was administered two weeks later. At termination, blood, spleen, and bone marrow was collected for analysis by flow cytometry. Surface expression of CD45, CD19, IgM, and IgG1 was assessed to determine white blood cell count, B cells, and pre- and post-class switch recombination (CSR), respectively. While no significant changes to B cell populations were observed in bone marrow or spleen, we demonstrate that CYT-0853 significantly decreases the median number of circulating CD45+ and IgG1 (post-CSR) B cells (61.8% vs. 31.6% and 8.7% vs. 4.4%, respectively). In addition, we observed a modest, significant increase in the amount of IgM+ (pre-CSR) B cells. These results were complemented by an associated overall significant decrease in circulating IgM levels. Of note, no adverse effects were observed in these mice over this treatment period. Based on these data and the role of B cells not only in antibody production, but also as antigen-presenting cells in multiple sclerosis, we tested our molecule in the myelin oligodendrocyte glycoprotein35-55-experimental autoimmune encephalomyelitis model of multiple sclerosis. Prophylactic treatment using 40mg/kg CYT-0853 did not affect disease activity or circulating cytokine production, however we observed a significant decrease in the spleen. Based on these results, further exploration is warranted to harness the power of CYT-0853 on the AID/RAD51 axis. This specific targeting may elicit beneficial therapeutic changes to B-lymphocyte populations and provide a novel immunomodulatory target to treat immunity and inflammation. Taken together, these data provide a foundation for continued preclinical development of CYT-0853 with applicability towards autoimmune diseases. Disclosures Aprahamian: Cyteir Therapeutics: Consultancy. Day:Cyteir Therapeutics: Employment. Mills:Cyteir Therapeutics: Employment, Equity Ownership.

Navigating the cutaneous B-cell lymphomas: avoiding the rocky shoals.

In recent years great progress has been made in understanding the classification of lymphomas. The integration of morphologic, clinical, immunophenotypic, and molecular features provides a rational basis for defining disease entities and has led to worldwide consensus. Hematopathologists and dermatopathologists have worked together to define those lymphomas that are present most commonly in the skin. Some cutaneous lymphomas have distinctive features and differ from their nodal counterparts. This is most evident in the delineation of primary cutaneous follicle center lymphoma and primary cutaneous marginal zone lymphoma. Both are very indolent, with low risk to spread beyond the skin. Primary cutaneous marginal zone lymphoma shows evidence of immunoglobulin class switching, as distinct from involvement by other extranodal marginal zone lymphomas of MALT type, which may involve the skin secondarily. Some have suggested that primary cutaneous marginal zone lymphoma may be considered a benign clonal expansion, probably driven by antigen. Many cutaneous lymphomas share biological and clinical features with their systemic counterparts. For example, primary cutaneous large B-cell lymphoma, leg type, exhibits a similar gene expression and molecular profile as diffuse large B-cell lymphoma of the activated B-cell type, especially for those cases arising in other extranodal sites. In addition, Epstein-Barr virus plays a role in many cutaneous lesions including mucocutaneous ulcer, plasmablastic lymphoma, and even some cases of marginal zone lymphoma. These EBV-driven conditions may present primarily in the skin, but also involve other mainly extranodal sites. Thus, it is evident that some cutaneous and systemic lymphomas are driven by common pathogenetic mechanisms, necessitating an integrated approach for the classification of lymphoma in all sites.

CERTAIN PARACLINICAL INDICATORS IN NEWBORN PATIENTS WITH SEPSIS, WHOSE PARENTS HAVE CONSTANTLY LIVED UNDER DIFFERENT ECOLOGICAL CONDITIONS

Xenobiotics can have various effects on the human body, and especially on the course of fetal development. Numerous researchers point out that the nature of the external environment can negatively affect the course of certain diseases in both children and adults. Neonatal sepsis is one of the most serious diseases in neonatological practice and in our opinion is a completely unexplained topic of the impact of adverse environmental factors on the characteristics of sepsis in newborns, depending on the environmental situation of their parents. The aim of the study was to investigate the features of individual indicators of a pair of clinical trials in children with neonatal sepsis, whose parents lived permanently in different environmental conditions. Material and methods. To study this goal, a comprehensive survey of 260 newborns who in 2015-2018 suffered from neonatal sepsis. The proposed environmental risk factor (ERF) was a group-forming feature of a comprehensive assessment of the long-term load on the body of parents of newborns of anthropogenic pollution of air, water and soil in the regions, taking into account the environmental situation in regional centers. Thus, the first clinical group (main) included patients with neonatal sepsis, whose parents lived permanently in months with ERF 2.0 and more and with unfavorable environmental characteristics of the regional center. The second group (comparison) was formed by newborns with sepsis, whose parents lived permanently in areas with a low risk of adverse effects of these environmental factors on their body (ERF&lt;2.0). Results and discussion. The content in the serum of patients with neonatal sepsis interleukins-6, -8.-10, Creactive protein, presepsin and procalcitonin reflected the activity of the systemic inflammatory response of the body to the infectious agent. It should be noted that patients of clinical group I were more likely to have a high serum content of interleukin-10, which has an anti-inflammatory effect. Thus, the content of this interleukin .35.0 pg / ml in the blood occurred in the newborn of the main group in 23.7 % of cases, and in children of the comparison group – in 18.1 % of cases. Along with a clearer definition of patients with group I high levels of anti-inflammatory interleukin-10, in newborns found a significant decrease in serum concentrations of immunoglobulins of classes A, G, M. Conclusions. The decrease in the level of these serum immunoglobulins is probably due to the immunosuppressive effect of xenobiotics on the fetus, whose mother was in the conditions of long-term action of xenobiotics. This, in turn, reduces the body's resistance of the newborn to infection and contributes to a more severe course of the infectious process. Probably, more severe manifestations of neonatal sepsis in neonates of the I clinical group are partly due to a combination of decreased immunoglobulin synthesis and increased production of interleukin-10.

Association of soluble CD40 levels with -1 C>T CD40 polymorphism and chronic kidney disease in systemic lupus erythematosus.

BackgroundCD40 is a transmembrane protein mainly expressed on the antigen‐presenting cells surface. CD40 plays a crucial role in immunoglobulin class switching and antibodies production. Genetic polymorphisms in the CD40 gene have been associated with increased risk of systemic lupus erythematosus (SLE) in several populations. This study aimed to evaluate the association of CD40 polymorphisms (−1 C > T, rs1883832 and 6,048 G > T, rs4810485) with SLE susceptibility, as well as with mRNA expression and soluble CD40 (sCD40) levels.MethodsThe study included 293 patients with SLE and 294 control subjects (CS). Genotyping was performed by PCR‐RFLP method. CD40 mRNA expression was determined by quantitative real‐time PCR, and ELISA quantified sCD40 levels.ResultsThe CD40 polymorphisms −1 C > T and 6,048 G > T were associated with SLE susceptibility. There was no difference between CD40 mRNA expression and CD40 polymorphisms. The sCD40 levels were lower in SLE patients with TT haplotype, whereas higher sCD40 levels were associated with damage and impaired renal function according to SLICC and KDIGO. The sCD40 levels were negatively correlated with eGFR.ConclusionThe CD40 gene polymorphisms increase the risk of SLE in the western Mexican population. The sCD40 levels are associated with −1 C > T polymorphism and chronic kidney disease.

Immune response to Ichthyophthirius multifiliis and role of IgT.

The parasitic ciliate Ichthyophthirius multifiliis causes white spot disease in freshwater fish worldwide. The theront penetrates external surfaces of the naïve fish where it develops into the feeding trophont stage and elicits a protective immune response both at the affected site as well as at the systemic level. The present work compiles data and presents an overall model of the protective reactions induced. A wide spectrum of inflammatory reactions are established upon invasion but the specific protection is provided by adaptive factors. Immunoglobulin IgT is involved in protection of surfaces in several fish species and is thereby one of the first adaptive immune molecules reacting with the penetrating theront. IgT producing lymphocytes occur in epithelia, dispersed or associated with lymphoid cell aggregations (skin epidermis, fins, gills, nostrils and buccal cavities) but they are also present in central immune organs such as the head kidney, spleen and liver. When theronts invade immunized fish skin, they are encountered by host factors which opsonize the parasite and may result in complement activation, phagocytosis or cell‐mediated killing. However, antibody (IgT, IgM and IgD) binding to parasite cilia has been suggested to alter parasite behaviour and induce an escape reaction, whereby specific IgT (or other classes of immunoglobulin in fish surfaces) takes a central role in protection against the parasite.

THE IMMUNOMODELING ACTIVITY OF BISMUTH CITRATE IN EXPERIMENT 1-CSTL AND LABORATORY OF INDUSTRIAL TOXICOLOGY

The aim of the study. To investigate in experimental conditions the effect of bismuth citrate on the immune system of laboratory animals.&#x0D; Material and methods. One-time sensitization of tadpoles (subcutaneously into the ear) was performed according to the method of O.G. Alekseeva, A.I. Petkevich. The degree of sensitization was established after skin tests. Determined the leucocyte blood formula, T and B lymphocytes in peripheral blood by the rosette method, immunoglobulins of class A, M, G in serum - by the method of radial immunodiffusion of globulines in agar Difco, circulating and extracellular complexes detection of the response of blood cells to the allergen "in vitro" - by the reaction of specific leukocyte lysis.&#x0D; Results. Bismuth citrate causes impaired immunological homeostasis in experimental animals. Indicators of nonspecific cellular component of the immune system have changed significantly, characterizing immunocomplex pathology. A decrease in the T-helper subpopulation was observed, indicating a significant immunomodulatory ability of bismuth citrate. At the same time, there was a significant increase in cells capable of rosette formation, which suggests that bismuth citrate is capable of provoking sensitization of the body. In the humoral immunity link, significant differences in the content of circulating immune complexes in sensitized and control animals were observed. Immunoglobulin levels were unchanged relative to control. The analysis of the sensitizing effect indicates that the most probable result of sensitization of the organism in this mode of receipt of bismuth citrate is the formation of type III hypersensitivity.&#x0D; Conclusions. Bismuth citrate under experimental conditions causes changes in the immune system of experimental animals, which are characteristic of the allergenic response of the organism to the complement-dependent type.

Formation of local protection of the respiratory tract in Holstein calves

The processes of formation of the local protection of the respiratory tract in nasal secretion samples of Holstein calves from birth to 6 months of age that were born from clinically healthy cows were examined. Only clinically healthy animals were involved in the experiment. Nasal secretion samples were collected with the help of a sterile foam tampon with a fixing thread in order to assess local immunity in animals at the age of 1, 5, 10, 30, 60, 90, 120, 150, and 180 days. Total protein, mucin, lysozyme activity, and alkaline phosphatase (ALP) were determined using commercially available methods. The amounts of immunoglobulins were studied using disk electrophoresis in polyacrylamide gel with commercial marker proteins. Studies have shown that the primary potential of local protection of the respiratory tract is formed during intrauterine development. The main factors of primary potential are mucin, lysozyme, alkaline phosphatase, and secretory immunoglobulins of classes A (SIgA) and M (SIgM), which provide safety of adaptation in newborns to the new conditions of existence. There is a decrease in mucin level by 47.3% (P < 0.001), in SIgA by a factor of 6.7 (P < 0.001), and in SIgM by a factor of 6 (P < 0.001) during the first week of life. However, the risk of immunodeficiency is mitigated by the preservation of relatively high levels of lysozyme, ALP, and IgG, creating optimal conditions for the processes of formation of respiratory functions in young animals during the first 2 months of life. After that, SIgA increases by 70% (P > 0.05) at the age of 60 days and mucin level increases by 56.7% (P < 0.001) between 2 and 6 months.

P1.04-18 Interleukin-5 Drives the Expansion of Pulmonary B-1 B Cells and Restricts Lung Tumor Growth

Therapies that target immune cells to treat lung cancer are becoming commonplace, highlighting the critical role of the immune system in the lung microenvironment. The lungs are host to a variety of immune populations, including B cells, whose primary function is to secrete antibodies that target foreign pathogens. Despite the presence of B cells in solid tumors and their prevalence in the lung, the role of B cells in lung cancer is largely undefined. We have found that transgenic mice which over-express the protein IL-5 (IL5Tg mice) have both an expansion of lung B cells and a decrease in lung tumor burden. IL-5 has previously been shown to induce the proliferation, recruitment, and antibody production of certain B cell subsets. Therefore, we hypothesize that IL-5-responsive B cells may decrease lung tumor growth. We used IL5Tg transgenic mice that over-express IL-5 to study the impact of B cells on tumor growth in the lungs. Lewis lung carcinoma (LLC) cells were injected intravenously (IV) to seed the lungs. Circulating serum antibodies were quantified by ELISA. Flow cytometry was used to quantify immune cell subsets in the lungs, as well as antibody binding to tumor cells and tumor cell cytotoxicity. Histological analysis was used to measure tumor burden. We found that the increase in lung B cells in IL5Tg mice was due to an increase in B-1 B cells which co-expressed the antibody class immunoglobulin M (IgM). Importantly, IL5Tg mice had a decrease in lung tumor burden compared to WT mice, as well as an increase in the total number of lung-infiltrating B cells compared to naive mice. Circulating IgM was elevated in IL5Tg mice compared to WT, and we found that these IgM antibodies were able to bind to LLC cells. Binding of IgM antibodies to the surface of target cells has previously been shown to induce cell death via the complement pathway, and preliminary data suggests that serum antibodies from IL5Tg mice can induce tumor cell cytotoxicity. We have found that excess IL-5 drives the expansion of lung B-1 B cells and decreases lung tumor burden, suggesting that IL-5 may promote anti-tumorigenic immune cell activities in the lungs. Illuminating the specific role of B cells in lung cancer growth will help deconvolute the complex interplay between host immune cells and malignant cells and could reveal new avenues for immunotherapy development.

Food-Specific IgE and IgG Antibodies in Patients With Chronic Rhinosinusitis With Nasal Polyps: A Case-Control Study.

EPOS 2012 states that investigation is needed to study a possible role for food allergy in the initiation and perpetuation of chronic rhinosinusitis with nasal polyps (CRSwNP). Our main goal was to compare serum levels of food-specific immunoglobulin G (IgG) and IgE antibodies in patients with CRSwNP and controls. A prospective case-control study with 33 patients with CRSwNP and 31 controls without CRS was carried out. Clinical data were gathered through a systematic interview and blood sample was collected. Enzyme-linked immunosorbent assay tests using OmegaDiagnostics kit with 40 food allergens for detection of specific IgG antibodies were performed and food-specific IgE antibodies were determined by immunoassay using ImmunoCAP. Immunoglobulin classes and IgG subclasses levels were also evaluated. Statistical analysis was performed using SPSS v.23. The overall sum of food IgG antibodies was significantly lower in CRSwNP compared to control group, and this difference was also observed for different specific IgG antibodies (corn, soya, grain legumes, pear and apple, berries, citric fruit). In controls, a positive correlation between IgG1 and the sum of food IgG antibodies was seen, but in CRSwNP group a negative correlation was found. In addition, a significant higher level of IgG1 and lower IgG2 and IgG3 was found among patients with CRSwNP. Levels of serum-specific IgE antibodies against multiallergen food mix (fx5) and against shrimp, strawberry, orange, rye, or egg yolk, as well as the sum of food IgE antibodies, did not differ significantly between the groups. These findings suggest that food allergy does not have an important role in CRSwNP etiopathogenesis, whether it is IgG or IgE mediated. Moreover, the observed suppression of specific IgG antibodies against food allergens, its negative correlation with IgG1 and the IgG1 switching in CRSwNP, can be related to deviated IgG responses against other targets (eg, airborne particles) and warrants future investigation.

The nuclear structural protein NuMA is a negative regulator of 53BP1 in DNA double-strand break repair.

P53-binding protein 1 (53BP1) mediates DNA repair pathway choice and promotes checkpoint activation. Chromatin marks induced by DNA double-strand breaks and recognized by 53BP1 enable focal accumulation of this multifunctional repair factor at damaged chromatin. Here, we unveil an additional level of regulation of 53BP1 outside repair foci. 53BP1 movements are constrained throughout the nucleoplasm and increase in response to DNA damage. 53BP1 interacts with the structural protein NuMA, which controls 53BP1 diffusion. This interaction, and colocalization between the two proteins in vitro and in breast tissues, is reduced after DNA damage. In cell lines and breast carcinoma NuMA prevents 53BP1 accumulation at DNA breaks, and high NuMA expression predicts better patient outcomes. Manipulating NuMA expression alters PARP inhibitor sensitivity of BRCA1-null cells, end-joining activity, and immunoglobulin class switching that rely on 53BP1. We propose a mechanism involving the sequestration of 53BP1 by NuMA in the absence of DNA damage. Such a mechanism may have evolved to disable repair functions and may be a decisive factor for tumor responses to genotoxic treatments.

A Computational Pipeline for the Diagnosis of CVID Patients.

Common variable immunodeficiency (CVID) is one of the most frequently diagnosed primary antibody deficiencies (PADs), a group of disorders characterized by a decrease in one or more immunoglobulin (sub)classes and/or impaired antibody responses caused by inborn defects in B cells in the absence of other major immune defects. CVID patients suffer from recurrent infections and disease-related, non-infectious, complications such as autoimmune manifestations, lymphoproliferation, and malignancies. A timely diagnosis is essential for optimal follow-up and treatment. However, CVID is by definition a diagnosis of exclusion, thereby covering a heterogeneous patient population and making it difficult to establish a definite diagnosis. To aid the diagnosis of CVID patients, and distinguish them from other PADs, we developed an automated machine learning pipeline which performs automated diagnosis based on flow cytometric immunophenotyping. Using this pipeline, we analyzed the immunophenotypic profile in a pediatric and adult cohort of 28 patients with CVID, 23 patients with idiopathic primary hypogammaglobulinemia, 21 patients with IgG subclass deficiency, six patients with isolated IgA deficiency, one patient with isolated IgM deficiency, and 100 unrelated healthy controls. Flow cytometry analysis is traditionally done by manual identification of the cell populations of interest. Yet, this approach has severe limitations including subjectivity of the manual gating and bias toward known populations. To overcome these limitations, we here propose an automated computational flow cytometry pipeline that successfully distinguishes CVID phenotypes from other PADs and healthy controls. Compared to the traditional, manual analysis, our pipeline is fully automated, performing automated quality control and data pre-processing, automated population identification (gating) and deriving features from these populations to build a machine learning classifier to distinguish CVID from other PADs and healthy controls. This results in a more reproducible flow cytometry analysis, and improves the diagnosis compared to manual analysis: our pipelines achieve on average a balanced accuracy score of 0.93 (±0.07), whereas using the manually extracted populations, an averaged balanced accuracy score of 0.72 (±0.23) is achieved.

Convergent Evolution of Mucosal Immune Responses at the Buccal Cavity of Teleost Fish.

SummaryThe buccal mucosa (BM) is a critical first line of defense in terrestrial animals. To gain further insights into the evolutionary origins and primordial roles of BM in teleosts here we show that rainbow trout, a teleost fish, contains a diffuse mucosal associated lymphoid tissue (MALT) within its buccal cavity. Upon parasite infection, a fish immunoglobulin specialized in mucosal immunity (sIgT) was induced to a high degree, and parasite-specific sIgT responses were mainly detected in the buccal mucus. Moreover, we show that the trout buccal microbiota is prevalently coated with sIgT. Overall our findings revealed that the MALT is present in the BM of a non-tetrapod species. As fish IgT and mucus-producing cells are evolutionarily unrelated to mammalian IgA and salivary glands, respectively, our findings indicate that mucosal immune responses in the BM of teleost fish and tetrapods evolved through a process of convergent evolution.

Efficiency of combined therapy of figuratum and parainfectious erythemas with the use of immunomodulators of different origins and MNRI

The aim of the study was to study the etiopathogenesis of erythema, comparative study of the clinical efficacy of combination therapy with the use of immunomodulators in erythema, the study of the dynamics of immunological parameters as a result of therapy.Materials and methods. The study included 215 adult patients with erythema. Before treatment and 1.5 months after therapy in patients have been studied: the expression of TLRs on MLPC and skin cells by flow cytometry with the use of the mat for TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 (Caltag Laboratories, USA) using a flow cytometer FC-500 (Beckman Coulter, USA); the subpopulation composition of peripheral blood lymphocytes; the levels of pro-inflammatory and regulatory cytokines in the serum by the method of solid-phase ELISA using test systems company «Biosource» (Austria); the main classes of immunoglobulins (IgG, IgM, IgA) in serum.Results. It was shown that these pathogenetic and diagnostic significance of viruses of the family nerpesviri-dae, bacterial contamination by different types of microorganisms (Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus spp., Proteus vulgaris). A substance containing antigens of conditionally pathogenic microorganisms, and the use of neuro sensorimotor reflex integration (MNRI) contributed to the decline in the number and severity of recurrences of erythema, the extension of the range expressed Toll-like receptors; increase of lymphocytes with immunophenotype CD3+, CD4+, CD25+, CD95+; increase in IFN-y, decreased IL-4; largely reduced the level of total IgE than other types of therapy.Conclusion. The inclusion in therapy of patients with erythema, immunomodulator of bacterial origin and MNRI promotes increase of clinical efficiency and correlates with the correction of immunological disorders.