Local Humoral Immunity Research Articles

Sexual dimorphism in fish innate immunity: A functional and transcriptional study in yellowtail kingfish

Sexual dimorphism in immunity has been extensively documented across vertebrates, with marked contrasts observed in immune responses between males and females. These variations are mainly attributed to oestrogens conferring enhanced immune responses in females, while males exhibit greater susceptibility to pathogens. However, in the light of the data, consensus is lacking, as different physiological and environmental factors such, as epigenetics, may impact sex-biased immunity. In fish, the regulation of immune responses through sex hormones is primarily determined by the leucocyte function, which contains sex steroid receptors. However, comparative sex-based research in fish immunity is still very limited. This study aimed to evaluate, for the first time, the disparities between males and females yellowtail kingfish (Seriola lalandi) juveniles in several parameters of local humoral innate immunity related to mucosae (skin mucus and foregut homogenates) and reproductive tissue (ovary and testis homogenates), as well as in serum. We investigated the sexual dimorphism in the expression patterns of genes coding for antimicrobial peptides, antiviral markers, and cytokines. Our findings revealed that the yellowtail kingfish males exhibit significantly higher levels of innate immune parameters, both functionally and transcriptionally, compared to females. These results suggest that females may have a higher susceptibility to pathogen infections, potentially leading to latent infections, which deservers further investigations. Understanding these sex-based differences in immunity could guide breeding strategies improvements and disease management in aquaculture facilities.

Imunoestimulação da resposta broncoalveolar de bezerros vacinados contra complexo respiratório bovino

ABSTRACT: Although intranasal bovine respiratory disease (BRD) vaccines containing live attenuated virus elicit greater stimulation of local humoral immunity response, they can mimic a viral infection, responsible for reducing innate defense during the establishment of vaccine-induced immunity. Probiotics containing Saccharomyces cerevisiae or Enterococcus faecium reduced the occurrence of BRD in neonatal calves challenged with the bovine respiratory syncytial virus (BRSV). Furthermore, the probiotics potentiated the humoral immune response after vaccination in murine models, raising the question of whether they could have the same effect in calves. This study aimed to verify if the probiotic containing E. faecium and S. cerevisiae attenuates the inflammation caused by the vaccine against BRD in the respiratory tract region in calves. Twenty-four healthy Jersey calves, aged 6 to 7 months old, were divided into the groups: control (C), supplemented (S), vaccinated (V), and supplemented and vaccinated (SV), with six animals in each of them. Supplemented groups (S and SV) received S. cerevisiae and E. faecium once a day on D-15 for 51 days (Probios precise®, Ouro Fino®, 2g/day/animal). Vaccinated groups (V and SV) received a single dose of an intranasal BRD vaccine on day 0 (Inforce®, Zoetis®, 1ml/ nostril). The control group was not supplemented or vaccined. Irritation of the respiratory tract and bronchoalveolar (BA) evaluations: cytology, phagocyte function, and IgA were measured on D-15, D3, D7, and D21. The vaccinated groups showed greater irritation of the nasopharynx and trachea. However, only Group V showed a reduction in BA phagocyte function and an increase in cellularity by a neutrophil influx in the BA region. Regarding IgA BA, SV showed the greatest increase, followed by S and V, concerning C. We conclude that isolated supplementation with E. faecium and S. cerevisiae promoted increased production of BA IgA. In association with the vaccine, the supplementation attenuated the inflammation of the respiratory tract produced by the vaccine itself, avoiding the reduction of phagocyte function BA, besides potentiating the humoral immune response of the vaccine containing live attenuated virus against BRD.

THE CONTENT OF SECRETORY IMMUNOGLOBULIN A IN BREAST MILK OF PATIENTS WITH LACTATIONAL MASTITIS

Introduction. The protective function of breast milk is provided by a number of specific and non-specific anti-infective factors: secretory immunoglobulin A, immunoglobulins A, G, M, E, cellular elements (neutrophils, lymphocytes, macrophages), cytokines, lactoferrin, lysozyme, oligosaccharides, bifido- and lactobacteria, antioxidants and.
 The main class of human milk immunoglobulins is secretory immunoglobulin A (sIgA), which protects the surfaces of mucous membranes from the penetration of microorganisms, neutralizes toxins and localizes viruses, and stimulates phagocytosis. Currently, information on the local level of secretory immunoglobulin A in the development of the inflammatory process in the mammary gland is not numerous and requires research.
 The aim of the study. To assess the content of secretory IgA in breast milk of patients with lactational mastitis.
 Research metods. The study dealt with breast milk of 97 women aged 18 to 36 (average age: 26±5 years old). The standard indicators of the secretory immunoglobulin A contents in 30 practically healthy lactating women of the same age – the control group – were the issue of the analyses. The first group included 30 women with lactostasis, the second group consisted of 37 women with lactational mastitis. The content of secretory immunoglobulin A was determined by enzyme immunoassay using an automatic STAT FAX 303 plus analyzer.
 Results and discussion. The concentration of sIgA in breast milk with lactostasis is on average 0.72±0.04 mg/ml, which is 3 times higher than the control indicator (0.24±0.02 mg/ml; p<0.05). During the development of lactational mastitis, the level of sIgA is 0.75±0.05 mg/ml, which exceeds the control values by 3.1 times (р<0.05). The content of sIgA in the breast milk of women with lactostasis does not differ from the level of this immunoglobulin in patients with lactational mastitis. A high concentration of sIgA indicates the activation of immune protection on mucous membranes.
 Conclusions. Increased production of secretory immunoglobulin A in breast milk was found in experimental groups compared to practically healthy women, which indicates activation of local humoral immunity as a reaction to infection.

Tissue-resident memory T cells trigger rapid exudation and local antibody accumulation

Adaptive immunity is didactically partitioned into humoral and cell-mediated effector mechanisms, which may imply that each arm is separate and does not function together. Here, we report that the activation of CD8+ resident memory T cells (TRM) in nonlymphoid tissues triggers vascular permeability, which facilitates rapid distribution of serum antibodies into local tissues. TRM reactivation was associated with transcriptional upregulation of antiviral signaling pathways as well as Fc receptors and components of the complement cascade. Effects were local, but evidence is presented that TRM in brain and reproductive mucosa are both competent to induce rapid antibody exudation. TRM reactivation in the mouse female genital tract increased local concentrations of virus-specific neutralizing antibodies, including anti-vesicular stomatitis virus, and passively transferred anti-HIV antibodies. We showed that this response was sufficient to increase the efficacy of ex vivo vesicular stomatitis virus neutralization. These results indicate that CD8+ TRM antigen recognition can enhance local humoral immunity.

Orally administered recombinant Lactobacillus expressing African swine fever virus antigens that induced immunity responses.

African swine fever (ASF) is a highly contagious, acute, febrile disease caused by the African swine fever virus (ASFV), with morbidity and mortality rates approaching 100% in domestic and wild swine, resulting in massive economic losses to the pig industry worldwide. This study aimed to express the p30, p54, and p72 proteins encoded by ASFV in vitro using the Lactobacillus lactis (L. lactis) expression system. Here, six new functional recombinant L. lactis were constructed, and the expression of the p30 protein, p54 protein, p72 protein, p30-LTB (heat-labile enterotoxin B, LTB) fusion protein, p54-LTB fusion protein, and the p72-LTB fusion protein was successfully detected by Western blot analysis. Following oral immunization of rabbits with recombinant L. lactis, serum IgG, intestinal mucosal sIgA, cytokines (IL-4 and INF-γ), and splenocyte viability were higher than in the control group via ELISA. Notably, without the LTB adjuvant group, humoral and Th1 cellular immunity were promoted, whereas, with the LTB adjuvant group, local mucosal immunity, humoral immunity, and Th2 cellular immunity were promoted, providing new insights into the design and development of an ASFV subunit vaccine.

The correlation of Th22 and regulatory T cells with Helicobacter pylori infection in patients with chronic gastritis.

Helicobacter pyloriis planted in the human stomach and is the most common cause of chronic gastritis, which produced specific local and systemic humoral immunity, while the associations of these immune responses and H. pylori in the development of chronic gastritis remain unclear. This study analyzed histology, the number of Th22 and regulatory T (Treg) cells, and the levels of inflammation- and gastritis-related indicators between 22 H. pylori-infected and 24 non-H. pylori-infected chronic gastritis patients by hematoxylin-eosinstaining, enzyme-linked immunosorbent assay, quantitative reverse transcription PCR, and flow cytometry analysis. This study found that the pathological damage degree of gastric mucosa in H. pylori infection patients was more serious. In the H. pylori-infected patient serum, the gastrin, G-17, interleukins(IL)-22, transforming growth factor(TGF)-β, tumor necrosis factor(TNF)-α, IL-4, and IL-17A levels were notably raised, while the interferon(IFN)-γ level was inhibited, and in gastric mucosa, and except IFN-γ, the IL-22, forkhead box P3 (Foxp3), TNF-α, IL-4, and IL-17A mRNA levels were raised too. The receiver operating characteristiccurve analysis indicates serum IL-22, TGF-β, TNF-α, IL-4, and IL-17A are suitable for differential diagnosis of H. pylori infection. In addition, in the peripheral blood, the percentages of the IL-22+ CD4+ and Foxp3+ CD4+ T cells were raised with H. pylori infection. The positive correlation between IL-22 and Foxp3 mRNA levels and the degree of H. pylori colonization and gastric mucositis by Pearson'scorrelation analysis. Treg and Th22 cells were positively associated with the degree of H. pylori infection and the severity of gastritis. In summary, this study provides an experimental basis for the study of the eradication of H. pylori and the biological mechanism of chronic gastritis.

The role of the palatine tonsils in the regulation of the state of humoral local immunity in patients with chronic inflammatory diseases of the nose and perinasal sinuses when infected with viruses of the respiratory group

Topicality: Acute respiratory diseases of the upper respiratory tract, which in the vast majority of cases are caused by viruses, especially with frequent recurrences of diseases, reduce local and system immunity, which often leads to the accumulation of bacterial infection due to the activation of endogenous microflora or exogenous infection and causes the development of viral-bacterial diseases. Recurrent respiratory diseases contribute to the formation of chronic pathology of the oro- and nasopharynx, which in turn supports the violation of the body's protective mechanisms. Previous studies have shown that the tonsils of the lymphadenoid pharyngeal ring perform the function of immunological protection, and the absence of palatine tonsils (PT) negatively affects the state of both local and system immunity. Aim: to determine the role of the palatine tonsils in the realization of local humoral reactions in patients with chronic inflammatory diseases of nose and paranasal sinuses when infected with viruses of the respiratory group. Materials and methods: Studies were conducted in 134 patients with chronic inflammatory diseases (CID) of nose and paranasal sinuses (PNS) in remission, among whom 93 had a diagnosis of chronic rhinosinusitis, 27 – chronic catarrhal rhinitis, 14 – allergic rhinitis with chronic bronchitis. The control group consisted of 47 practically healthy persons without pathology from the ENT organs. The examined persons were divided into those who had PT and those from whom they were surgically removed no earlier than five years ago. All examinees were divided into those who had anamnestic antibodies in their blood to one or more of the respiratory viruses, such as: adenovirus, respiratory syncytial virus and influenza A virus in a clinically significant titer, and those in whom antibodies were not detected. The local humoral immunity state of patients was assessed by the content of immune complexes (IC) and concentrations of secretory immunoglobulin A (sIgA), interferons alpha- (α-IFN) and gamma- (γ-IFN) in the oropharyngeal secretion (OS), which were determined by the immunoenzymatic method using sets of reagents for enzyme-linked immunosorbent assays ("Cytokin", RF) and the Stat Fax-2100 analyzer (USA). All examined patients had a healthy state of the oral cavity. Results: The presence or absence of palatine tonsils changes the levels of number of humoral factors of local immunity in oropharyngeal secretion in practically healthy donors and in patients with chronic inflammatory diseases of the nose and PNS. The production of sIgA and α-IFN in OS depends on PM, decreasing in practically healthy persons of the control group who underwent tonsillectomy. Chronic inflammatory diseases of the nose and PNS caused a decrease in the concentration of IC and α-IFN in the OS of patients of both groups- with or without PT. In patients with normal PMs who got sick with viral infections, in contrast to those with removed PMs, α-IFN level in OS increased and practically did not differ from those in healthy donors. The greatest suppression of α-IFN production was determined in patients with CID of the nose and PNS with removed PM and with the absence of clinically significant titers of antiviral antibodies. The level of γ-IFN production did not reliably change in the OS of patients with CID of nose and PNS in the remission phase both with normal PT and after tonsillectomy, but depended to a greater extent on the transferred infectious disease caused by viruses of the respiratory group. Conclusion: PTs play a significant role in the formation of a number of reactions of local immunity in the naso- and oropharynx, both in practically healthy individuals and in patients with chronic inflammatory disease of nose and paranasal sinuses. Therefore, the determination of local immunity indices in patients with chronic inflammatory diseases of the upper respiratory tract with different status of the palatine tonsils, who are in contact or had contact with respiratory viruses, provides an opportunity to identify the most vulnerable links in the system of their anti-infective protection, which should first of all be paid attention to during examination and treatment of patients with chronic inflammatory diseases of the upper respiratory tract, especially in cases of frequent recurrence of acute respiratory diseases.

Antimicrobial effect of nitric oxide

138 patients who were diagnosed with gastric and duodenal ulcers during EGDS were treated. The inclusion of exogenous nitric oxide in the anti-ulcer treatment of patients contributed to the eradication of Hp, restoration of the membrane component of damaged cells, stabilization of local immune reactions and humoral immunity. NO-therapy accelerated the epithelialization of ulcers by 2 times compared to patients who did not receive nitric oxide in complex therapy.

Natural SARS-CoV-2 Infection Affects Neutralizing Activity in Saliva of Vaccinees.

BackgroundSARS-CoV-2 transmission mainly occurs through exposure of the upper airway mucosa to infected secretions such as saliva, which are excreted by an infected person. Thus, oral mucosal immunity plays a central role in the prevention of and early defense against SARS-CoV-2 infection. Although virus-specific antibody response has been extensively investigated in blood samples of SARS-CoV-2-infected patients and vaccinees, local humoral immunity in the oral cavity and its relationship to systemic antibody levels needs to be further addressed.Material and MethodsWe fine-tuned a virus neutralization assay (vNTA) to measure the neutralizing activity (NA) of plasma and saliva samples from 20 SARS-CoV-2-infected (SI), 40 SARS-CoV-2-vaccinated (SV), and 28 SARS-CoV-2-vaccinated subjects with a history of infection (SIV) using the “wild type” SARS-CoV-2 lineage B.1 (EU) and the Delta (B.1.617.2) strains. To validate the vNTA results, the presence of neutralizing antibodies (NAbs) to the spike receptor binding domain (RBD) was evaluated with an ELISA assay.ResultsNA to SARS-CoV-2 lineage B.1 (EU) was present in plasma samples from all the tested subjects, with higher titers in SIV compared to both SI and SV. Conversely, NA was detected in saliva samples from 10.3% SV, 45% SI, and 92.6% SIV, with significantly lower titers in SV compared to both SI and SIV. The detection of NAbs in saliva reflected its reduced NA in SV.DiscussionThe difference in NA of plasma vs. saliva was confirmed in a vNTA where the SARS-CoV-2 B.1 and Delta strains were tested head-to-head, which also revealed a reduced NA of both specimens compared to the B.1 variant.ConclusionsThe administration of SARS-CoV-2 vaccines was associated with limited virus NA in the oral cavity, as measured in saliva and in comparison to plasma. This difference was more evident in vaccinees without a history of SARS-CoV-2 infection, possibly highlighting the importance of local exposure at the site of virus acquisition to effectively prevent the infection and block its spread. Nevertheless, the presence of immune escape mutations as possibly represented by the SARS-CoV-2 Delta variant negatively affects both local and systemic efficacy of NA associated with vaccination.

The influence of parenteral influenza vaccination on local immunity indices and microbiota of oropharyngeal secretion in patients with chronic inflammatory diseases of the upper respiratory tract

When studying the effect of vaccines against influenza, attention is mainly paid to obtaining high titers of protective antibodies in the blood and reducing the incidence of respiratory infections among vaccinated people. At the same time, the changes occurring in patient,s body from the factors of local specific and innate immunity remain insufficiently studied. The aim of the study was to determine the effect of parenteral influenza vaccination on the state of local immunity, cytology and microbiota of oropharyngeal secretion (OS) in patients with chronic inflammatory diseases of the upper respiratory tract. The study of immunological and microbiological parameters was performed in 32 patients with chronic inflammatory diseases of the upper respiratory tract, including 11 diagnosed with chronic rhinosinusitis, 9 – with chronic tonsillitis, 12 – with chronic pharyngitis, 3 and 12 weeks after vaccination with trivalent inactivated influenza-vaccine (PASTEUR, SA, France), which was administered intramuscularly. Single vaccination against influenza A and B has been shown to normalize reduced local humoral immunity indices, in particular sIgA and immune complexes concentrations, increase lymphocyte output to oropharyngeal secretions and cause a significant decrease in the representation of OS transient microflora without affecting the overall level of bacterial contamination. In both periods after the vaccination the reduced content of interferon-α in the OS of patients with chronic inflammatory diseases of the upper respiratory tract did not change. The obtained data allow to recommend vaccination against influenza virus in the period up to 3 months before the the beginning of mass infections as an effective means of stimulating the protective reactions of local immunity of oropha­rynx and nasopharynx mucous membranes in patients with chronic inflammatory diseases of the upper respiratory tract.

The phenotypic and functional study of tissue B cells in respiratory system provided important information for diseases and development of vaccines

The field of tissue‐resident B cells has received increasing attention, yet the feature of tissue B cells in respiratory system is unclear. Here, we first show that non‐circulating B cells obtained from nasal, trachea and lung tissues are numerically and phenotypically distinct from their circulating counterparts. Analysis of single cell transcriptome sequence identified multiple differentially expressed genes between non‐circulating B cells and circulating B cells, which illustrated their heterogeneity. Furthermore, we found high expression of CXCR3 on non‐circulating B cells, and the chemokine CXCL11 was also up‐regulated in the respiratory tissues, suggesting that CXCR3‐CXCL11 axis might accelerate the local resident of non‐circulating B cells in respiratory tract. Interestingly, intranasal immunization with BCG in mice elicited a sustained humoral immune response via induction of IgA and IgG Abs, which revealed the role of B cells. Meanwhile, tissue‐resident B cells, IgA+ and IgG+ memory B cells (MBCs) in respiratory tissues, as well as plasma cells in bone marrow, were expanded and maintained, and these subsets probably developed into antibody‐producing cells to participate in the local humoral immunity. Our data illustrate the phenotype and function of tissue B cells in the upper and lower airways, provide references for the prospective development of vaccines.

FUNCTIONAL ACTIVITY CHANGES OF THE MONONUCLEAR PHAGOCYTE SYSTEM IN PATIENTS WITH RAPIDLY PROGRESSIVE GENERALIZED PERIDONTITIS WITH PERIODONTAL MICROORGANISMS SENSITIVE AND RESISTANT TO DRUGS

Purpose. To reveal peculiarities of the mononuclear phagocyte functional activity in patients with rapidly progressive generalized periodontitis depending upon colonization of periodontal tissue by causative agents, sensitive and resistant to drugs, on the basis of studying activated blood monocytes, phagocytic activity of mononuclear phagocytes and production of the main classes of immunoglobulins, cytokine profile state. Material and methods. The investigation was carried out on 74 patients with rapidly progressive generalized periodontitis I-II and II-III severity, aged 35 to 50 years. Complex of the laboratory studies included immunophenotypic changes in the subpopulation composition of the mononuclear blood cells (CD3 +, CD4 +, CD8 +, CD14 +, CD16 +, CD25 +, HLA-DR and CD95+), phagocytic activity of neutrophils, level of cytokine production and synthesis of sIgA, IgA, IgG and IgM in the oral fluid. Results. On the basis of the conducted complex clinical-laboratory data it has been established that a significant decrease of the functional activity of the mononuclear phagocyte system stipulates the formation of the severe secondary deficiency of the cellular and humoral immunity in patients who are carries of the resistant strains of periodontal pathogenic agents of the rapidly progressive generalized periodontitis and moderate - at colonization of the periodontal complex with causative agents sensitive to drugs. The pathogenetic dependence of the drug sensitivity upon the nature of the disorders of the systemic and local humoral immunity has been shown: at the marked immunodeficiency the development of the bacterial resistance to drugs is possible, and preservation of the drug sensitivity to the causative agents of the rapidly progressive generalized periodontitis is associated with moderate disorders of anti-infectious mechanisms of the immune response. Conclusion. The assessment of the functional activity of the mononuclear phagocyte system, based on the study of the state of immunophenotypic changes in the subpopulation composition of mononuclear blood cells, phagocytic activity of monocytes, the content of immunoglobulins and cytokines in the oral fluid, gives valuable information to clarify the mechanisms of the development of various sensitivity of the periodontal-pathogenic flora of the periodontal tissue to protocol antibacterial therapy in the patients with rapidly progressive generalized periodontitis.

Treatment of aggressive (rapidly progressing) generalized periodontitis using systemic enzyme therapy in combination with osteoinductive medicines

The modern treatment strategy for various forms of generalized periodontitis includes the use of a wide arsenal of drugs, which in turn leads to an excessive drug load on the body, this actualizes the search and use of drugs that have a systemic effect. The aim of the study is to develop and study the effectiveness of the use of systemic enzyme therapy in the treatment of aggressive (rapidly progressing) generalized periodontitis. The work is based on the results of comprehensive clinical and laboratory studies of 61 patients with an aggressive form of generalized periodontitis, without concomitant pathologies (30 main groups and 31 comparison groups). A control group consisted of 20 healthy donor volunteers. We used standard clinical, paraclinical, laboratory diagnostic methods, supplemented by dental volumetric tomography. In patients of both observation groups, standard protocol therapy was used, in the comparison group supplemented pathogenic therapy, and in the main group – multienzyme drug (Wobenzym) and rhBMP-2 injections. A comparative analysis of clinical, paraclinical, and laboratory studies has shown that the use of systemic enzyme therapy supplemented with recombinant morphogenetic protein (rhBMP-2) in patients with aggressive (rapidly progressing) generalized periodontitis is comparable to that of standard polymedicine therapy supplemented with pathogenic therapy, and leads to a faster control (average 13.7±0.7 days) of an active inflammatory and destructive process, normalization of indices of local humoral immunity, cytokine profile, free radical oxidation and antioxidant defense system, achievement of clinical and radiological remission in more than 90% of patients.

Сучасний погляд на проблему рецидивних гіперпластичних процесів ендометрія

A review of current data on the problem of recurrent endometrial hyperplasia, presented in modern domestic and foreign literature, its place in the structure of gynecological morbidity, the impact on reducing fertility among women of childbearing age and the growth of cancer. The issues of existing classifications, their relevance and ease of use are considered. The main etiological factors and clinical and anamnestic factors of endometrial hyperplastic processes, prognostic influence of urogenital infections on endometrial pathology are highlighted. Possible mechanisms of pathogenesis are described, including disturbances of hormonal regulation, factors of local humoral and cellular immunity and molecular mechanisms of cellular regulation. The important role of biomolecular markers in the pathogenesis of hyperplastic processes and the correspondence of their degree of activity to morphological changes of the endometrium, because understanding the basics of induction of cell growth, especially in tumor tissue transformation, is an integral part of a competent approach to the management of patients with hyperplasia. The role of genetic research to assess the risk of malignancy and, accordingly, the choice of conservative or operative treatment tactics is considered. Modern morphometric diagnostic criteria are presented. Emphasis is placed on the need to collect endometrial biopsies under the control of a hysteroscope as the gold standard in the diagnosis of endometrial conditions. A separate role in the diagnosis at the present stage is given to immunohistochemical method; conducting not only histological examination of the removed endometrium, but also its receptor activity provides the ability to select the necessary pharmacological correction and predict the development of neoplastic changes. Variants of hormone therapy schemes of endometrial hyperplastic processes are described, and the order of their appointment, depending on the woman’s reproductive plans, concomitant diseases, age and constitutional features. Particular attention is paid to women with metabolic syndrome and the feasibility of including the correction of metabolic disorders in the treatment of such patients. The duration and frequency of observation of women during treatment are indicated. The presented data demonstrate the lack of a clear algorithm in the tactics of diagnosis and treatment of recurrent hyperplastic processes, which determines the relevance of in-depth scientific study in this area. Keywords: endometrial hyperplasia, recurrence, hysteroscopy, endometrial receptors

Dynamics of humoral immunity in gingival fluid during combined immunotherapy in pediatric patients with exacerbation of chronic granulomatous periodontitis

Chronic granulating periodontitis among children has a high prevalence, which determines the relevance of its research, medical and social significance. The exacerbation of the disease is accompanied by various disorders in the immune system and, above all, at the local level. However, changes in local humoral immunity in children with exacerbation of chronic granulating periodontitis (CGP) have not been studied. Impact of modern immunomodulators and, in particular, Derinat, on pathological changes of local parameters of humoral immunity was not analyzed in pediatric patients with this disorder.The aim of this work is to study local humoral immunity parameters during CGP exacerbation in children and on the background of combined therapy with Derinat, Unidox-Solutab, and Tycveolum (oil extract from pumpkin seeds).The analysis of immunological disorders in gingival fluid was performed in 93 children aged 12-14 years with exacerbation of chronic granulating periodontitis, who underwent examination and treatment in Belgorod city pediatric dental clinic, and at the Department of Pediatric Dentistry of Belgorod State National Research University in 2013-2015. The patients included in the study were divided into 2 groups: control group (n = 48), receiving conventional treatment, and the main group (n = 45), who, in addition to standard therapy, were treated with Derinat, Unidox-Solutab and Tycveolum. Contents of immunoglobulins in gingival fluid were determined by turbidimetric method using the Microlab-300 photometer. Lysozyme and lactoferrin were studied by solid-phase enzyme immunoassay.Exacerbation of chronic granulating periodontitis in childhood is associated with a representative decrease of sIgA, IgA, lysozyme and lactoferrin in gingival fluid, and increased levels of IgM and IgG. Upon treatment with innovative therapy, including derinate, as one of the components, in CGP patients, there was a significant improvement of immune parameters on the 14th day of observation. Normalization of all studied parameters of local immunity was noted in the patients of main group by the 30 th day of observation. Meanwhile, only some immunological parameters reached normal reference values in the control group of patients. Conclusion. The identified new mechanisms of Derinat effects upon humoral immunity and it’s effective use during exacerbation of chronic granulomatous periodontitis in children.

Optimisation of diagnostics and complex treatment of multi-form exudative erythema of cavity mucosa

The article presents the results of studies of complex treatment oral mucosa of patients with Erythema multiforme in detail of the developed algorithm. Subjective and objective criteria of oral mucosa state and local humoral immunity are proposed for evaluation of therapy effectiveness in patients with multi-form exudative erythema in the main and comparison group. A comparative characteristic of dental indicators of quality of life was made according to the questionnaire OHIP-14-RU in both groups.

Clinical and morphological features of allergic enterocolitis in young children

The purpose was to study the clinical and morphological features of allergic enterocolitis in young children.Material and methods. 29 patients aged from 2 months to 2 years with severe allergic enterocolitis, who underwent endoscopic examination with targeted biopsy of the intestinal mucosa, were treated. For comparison of morphological, histochemical and immunohistochemical features, intestinal mucosal biopsies of children of the same age with gastrointestinal disorders, caused by protracted or chronic course of non-specific non-ulcer process in the intestine, were used. Immunohistochemical studies were performed on serial paraffin sections in accordance with standard protocols using Ab68 (Macrophage Marker) Ab-3 (Clone KP1), IgA (Heavy Chain) Ab-2, IgE (Epsilon-Heavy Chain) Ab-1 monoclonal antibodies and Imaging Ultra Vision Quanto Detection System HRP DAB (USA).Results. The debut of gastrointestinal symptoms in young children with allergic enterocolitis was observed in the first months of life and was characterized by vomiting or persistent regurgitations, intense griping, diarrhea with large amounts of mucus and/or blood, and delayed physical development. Presence of inflammatory process of intestinal mucosa with eosinophilic infiltration was morphologically established. According to immunohistochemical study, expression of CD68-macrophages and increased expression of IgA and IgE in the duodenal mucosa were detected.Conclusions. The diagnosis of allergic enterocolitis at an early age is complicated by the absence of specific clinical gastrointestinal symptoms and non-informative allergic examination (IgE-independent mechanism of inflammation), which may require morphological study to verify the diagnosis. Morphologically, in the examined patients, not only the presence of inflammatory process of the mucous membrane of the intestine with expressed eosinophilic infiltration is established, but also the immunohistochemistry data revealed the increased expression of CD68-macrophages, IgA and IgE in the mucous membrane of the duodenum, indicating the activation of both cellular and local humoral immunity.

Electroporation of a nanoparticle-associated DNA vaccine induces higher inflammation and immunity compared to its delivery with microneedle patches in pigs

DNA vaccination is an attractive technology, based on its well-established manufacturing process, safety profile, adaptability to rapidly combat pandemic pathogens, and stability at ambient temperature; however an optimal delivery method of DNA remains to be determined. As pigs are a relevant model for humans, we comparatively evaluated the efficiency of vaccine DNA delivery in vivo to pigs using dissolvable microneedle patches, intradermal inoculation with needle (ID), surface electroporation (EP), with DNA associated or not to cationic poly-lactic-co-glycolic acid nanoparticles (NPs). We used a luciferase encoding plasmid (pLuc) as a reporter and vaccine plasmids encoding antigens from the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), a clinically-significant swine arterivirus. Patches were successful at inducing luciferase expression in skin although at lower level than EP. EP induced the cutaneaous recruitment of granulocytes, of MHC2posCD172Apos myeloid cells and type 1 conventional dendritic cells, in association with local production of IL-1β, IL-8 and IL-17; these local responses were more limited with ID and undetectable with patches. The addition of NP to EP especially promoted the recruitment of the MHC2posCD172Apos CD163int and CD163neg myeloid subsets. Notably we obtained the strongest and broadest IFNγ T-cell response against a panel of PRRSV antigens with DNA + NPs delivered by EP, whereas patches and ID were ineffective. The anti-PRRSV IgG responses were the highest with EP administration independently of NPs, mild with ID, and undetectable with patches. These results contrast with the immunogenicity and efficacy previously induced in mice with patches. This study concludes that successful DNA vaccine administration in skin can be achieved in pigs with electroporation and patches, but only the former induces local inflammation, humoral and cellular immunity, with the highest potency when NPs were used. This finding shows the importance of evaluating the delivery and immunogenicity of DNA vaccines beyond the mouse model in a preclinical model relevant to human such as pig and reveals that EP with DNA combined to NP induces strong immunogenicity.

Role of cytokines in pathogenesis of H. pylori induced gastric mucosal inflammation.

Helicobacter pylori gastritis is characterized by colonize the stomach and induce strong,specific local and systemic humoral and cell-mediated immunity. The aim of this study was toevaluate the level of several cytokines and their role in the pathogenesis of H. pylori gastritisand whether H. pylori stimulate the release of these cytokines. The levels of TH1 (IFN-γ andTNF-α) and TH2 (IL-4, IL-6 and IL-10) cytokines, as well as, inflammatory cytokines (IL-1αand IL-8) were determined. Most of the investigated cytokines (IL-1α, IL-6, IL-8, IL-10, IFNγ,TNF-α and GM-CSF) showed a significantly increased level in the sera of H. pylori gastricpatients as compared to subjects (control) with no infection which showed normal seruminterleukin levels. While IL-10 showed significant decrease when compared to subjects.These results conclude the pro-inflammatory cytokines ave a central role in thepathogenesis of H. pylori-induced mucosal inflammation.

The effect of diatomaceous earth in live, attenuated infectious bronchitis vaccine, immune responses, and protection against challenge

Live virus vaccines are commonly used in poultry production, particularly in broilers. Massive application and generation of a protective local mucosal and humoral immunity with no adverse effects is the main goal for this strategy. Live virus vaccines can be improved by adding adjuvants to boost mucosal innate and adaptive responses. In a previous study we showed that diatomaceous earth (DE) can be used as adjuvant in inactivated vaccines. The aim of this study was to test DE as adjuvant in an Ark-DPI live infectious bronchitis virus (IBV) vaccine after ocular or spray application. Titrating the virus alone or after addition of DE showed that DE had no detrimental effect on the vaccine virus. However, adding DE to the vaccine did not induce higher IgG titers in the serum and IgA titers in tears. It also did not affect the frequency of CD4+ T cells, CD8+ T cells and monocytes/macrophages in the blood and the spleen determined by flow cytometry. In addition, protection generated against IBV homologous challenges, measured by viral load in tears, respiratory signs and histopathology in tracheas, did not vary when DE was present in the vaccine formulation. Finally, we confirmed through our observations that Ark vaccines administered by hatchery spray cabinet elicit weaker immune responses and protection against an IBV homologous challenge compared to the same vaccine delivered via ocular route.