Characterization of HPV6/11-reactive T-cell subsets in papillomas of patients with juvenile-onset recurrent respiratory papillomatosis and identification of HPV11 E7-specific candidate TCR clonotypes.

Juvenile-onset recurrent respiratory papillomatosis (JORRP) is caused by persistent infection of epithelial cells by low-risk human papillomavirus (HPV) types 6 and 11. While multiple infiltrated immune cells have been reported to mediate disease progress, knowledge of HPV-reactive T-cell subsets in papillomas remains elusive. Through single-cell RNA sequencing and RNA microarray, we found that CD8+ tissue-resident memory T (CD8+ TRM) cells with strong interferon-gamma (IFN-γ) production expanded, and were negatively correlated to the disease severity in the frequency of surgery. These IFN-γ+ CD8+ memory T cells were readily activated and expanded in vitro by autologous dendritic cells loaded with HPV11 E7 peptide pool. Moreover, T cell receptor (TCR) clonal expansion was observed in JORRP papilloma tissues, indicating a biased TCR repertoire toward HPV-specific recognition. Finally, we identified and characterized HPV11 E7-specific candidate TCR clonotypes from IFN-γ+ CD8+ memory T cells, suggesting their potential application in TCR-engineered T cells (TCR-T) therapy for HPV11-related diseases. Our findings provided insights into the specific local immune response to HPV6/11 infection and highlighted the importance of IFN-γ+ CD8+ TRM cells in anti-HPV6/11 T-cell immunity.IMPORTANCEThe persistent recurrence of human papillomavirus (HPV) 6/11 infection in papillomas underscores the failure of local immune responses in patients with juvenile-onset recurrent respiratory papillomatosis (JORRP). Our previous study demonstrated that T cells constitute the predominant immune cell population in JORRP papilloma tissues. Understanding the T-cell-mediated immune responses within JORRP papilloma tissues is crucial for disease control. In the present study, we characterized CD8+ tissue-resident memory T (CD8+ TRM) cells as the primary T-cell subset responsible for local anti-HPV6/11 immunity. Moreover, we identified two HPV11 E7-specific candidate T cell receptor (TCR) clonotypes out of IFN-γ+ CD8+ memory T cells. Overall, our findings provided insights into the local immune responses to HPV6/11 infection and offered information for developing more effective immunotherapeutic strategies against JORRP.

Uncovering the Bronchoalveolar Single-Cell Landscape of Patients With Pulmonary Tuberculosis With Human Immunodeficiency Virus Type 1 Coinfection.

BACKGROUND: Coinfection of human immunodeficiency virus type 1 (HIV-1) is the most significant risk factor for tuberculosis (TB). The immune responses of the lung are essential to restrict the growth of Mycobacterium tuberculosis and avoid the emergence of the disease. Nevertheless, there is still limited knowledge about the local immune response in people with HIV-1-TB coinfection. METHODS: We employed single-cell RNA sequencing (scRNA-seq) on bronchoalveolar lavage fluid from 9 individuals with HIV-1-TB coinfection and 10 with pulmonary TB. RESULTS: A total of 19 058 cells were grouped into 4 major cell types: myeloid cells, T/natural killer (NK) cells, B cells, and epithelial cells. The myeloid cells and T/NK cells were further divided into 10 and 11 subsets, respectively. The proportions of dendritic cell subsets, CD4+ T cells, and NK cells were lower in the HIV-1-TB coinfection group compared to the TB group, while the frequency of CD8+ T cells was higher. Additionally, we identified numerous differentially expressed genes between the CD4+ and CD8+ T-cell subsets between the 2 groups. CONCLUSIONS: HIV-1 infection not only affects the abundance of immune cells in the lungs but also alters their functions in patients with pulmonary TB.

Adenoviral-vectored universal influenza vaccines administered intranasally reduce lung inflammatory responses upon viral challenge 15 months post-vaccination.

IMPORTANCE: Vaccines targeting highly conserved proteins can protect broadly against diverse viral strains. When a vaccine is administered to the respiratory tract, protection against disease is especially powerful. However, it is important to establish that this approach is safe. When vaccinated animals later encounter viruses, does reactivation of powerful local immunity, including T cell responses, damage the lungs? This study investigates the safety of mucosal vaccination of the respiratory tract. Non-replicating adenoviral vaccine vectors expressing conserved influenza virus proteins were given intranasally. This vaccine-induced protection persists for at least 15 months. Vaccination did not exacerbate inflammatory responses or tissue damage upon influenza virus infection. Instead, vaccination with nucleoprotein reduced cytokine responses and histopathology, while neutrophil and T cell responses resolved earlier. The results are promising for safe vaccination at the site of infection and thus have implications for the control of influenza and other respiratory viruses.

Clonal evolution and stereotyped sequences of human IgE lineages in aeroallergen-specific immunotherapy.

BACKGROUND: Allergic disease reflects specific inflammatory processes initiated by interaction between allergen and allergen-specific IgE. Specific immunotherapy (SIT) is an effective long-term treatment option, but the mechanisms by which SIT provides desensitization are not well understood. OBJECTIVE: Our aim was to characterize IgE sequences expressed by allergen-specific B cells over a 3-year longitudinal study of patients with aeroallergies who were undergoing SIT. METHODS: Allergen-specific IgE-expressing clones were identified by using combinatorial single-chain variable fragment libraries and tracked in PBMCs and nasal biopsy samples over a 3-year period with antibody gene repertoire sequencing. The characteristics of private IgE-expressing clones were compared with those of stereotyped or "public" IgE responses to the grass pollen allergen Phleum pratense (Phl p) 2. RESULT: Members of the same allergen-specific IgE lineages were observed in nasal biopsy samples and blood, and lineages detected at baseline persisted in blood and nasal biopsy samples after 3 years of SIT, including B cells that express IgE. Evidence of progressive class switch recombination to IgG subclasses was observed after 3 years of SIT. A common stereotyped Phl p 2-specific antibody heavy chain sequence was detected in multiple donors. The amino acid residues enriched in IgE-stereotyped sequences from seropositive donors were analyzed with machine learning and k-mer motif discovery. Stereotyped IgE sequences had lower overall rates of somatic hypermutation and antigen selection than did single-chain variable fragment-derived allergen-specific sequences or IgE sequences of unknown specificity. CONCLUSION: Longitudinal tracking of rare circulating and tissue-resident allergen-specific IgE+ clones demonstrates persistence of allergen-specific IgE+ clones, progressive class switch recombination to IgG subtypes, and distinct maturation of a stereotyped Phl p 2 clonotype.

INDICATORS OF DENTAL HEALTH AND LOCAL IMMUNITY IN YOUNG ADULTS WHO HAVE SUFFERED FROM CORONAVIRUS INFECTION.

OBJECTIVE: The aim: To determine the dental status and state of local immunity in young adults who have suffered from the coronavirus disease. PATIENTS AND METHODS: Materials and methods: The main group consisted of 30 people aged 20-22 years, who suffered from the coronavirus infection Covid19 6.1±1.2 months ago. The comparison group included 20 people who did not have a coronavirus infection. The control group consisted of 35 people, randomized by age and sex, who did not have signs of caries and periodontal tissue disease and did not have coronavirus disease. All patients were examined for dental status and local immunity. RESULTS: Results: The analysis of indicators of dental status revealed the possibility of the existence of a relationship between the signs of acute SARS-Cov2 viral infection and the development of caries and periodontal tissue diseases. Significant changes in the local immunity of the oral cavity were found in the examined patients, which had a pathogenetic influence on the development and progression of caries and periodontal tissue diseases: a significant increase in the level of Ig G, as well as a probable decrease in the concentration of SIg A relative to the comparison group, a probably higher normative value of pathogenic small- and medium-molecular CICs with a significant decrease in the level of physiological large-sized CICs relative to the comparison group, a decrease in the content of anti-inflammatory IL-4, as well as increased concentration of pro-inflammatory cytokines. CONCLUSION: Conclusions: Young adults who have suffered a coronavirus infection during the last 6 months have significantly higher caries prevalence, bleeding index, PMA index and hygiene index, halitosis, which indicates deeper tissue damage and tooth pathology with the formation of dentition defects than in the comparison group. Indicators of local immunity of the oral fluid have a deep and specific character.

Importance of antimicrobial factors for microbiocenosis and local immunity of the oral cavity in children with mucoviscidosis.

The involvement of commensals and opportunistic pathogens and the role of protective mechanisms in the development of dental diseases in children with cystic fibrosis require more detailed study. AIM: The aim of the study was to determine the ecological characteristics of the oral microbiota and some antimicrobial factors of saliva in children with mucoviscidosis. MATERIALS AND METHODS: The study involved an assessment of oral microbiota as complex ecological system that protects the human body from colonization by pathogenic flora in children with cystic fibrosis. Bacteriological studies have been performed on clinical material from 30 children with mucoviscidosis diagnosed with dental and periodontal diseases. RESULTS: In the microbiological study of plaque microbiota, 70 strains of opportunistic pathogens were isolated in patients with mucoviscidosis. The most significant were alpha-hemolytic Streptococci (40%). The proportion of bacteria of Neisseria genus in patients with cystic fibrosis was lower and amounted to 24.3%. C. albicans fungi were isolated in comparable values (18.5%), S. aureus (8.5%), as well as gram-negative strains of E. aerogenes (4.3%) and E. coli (4.3%) significantly dominated. The results indicate that opportunistic pathogens S. aureus, E. aerogenes and E. coli partially replaced the representatives of the normal oral microbiota alpha-hemolytic streptococci and non-pathogenic species of Neisseria genus in patients with mucoviscidosis. CONCLUSIONS: Microbiota of plaque in children with mucoviscidosis is characterized by an expansion of the spectrum of opportunistic pathogens due to Staphylococcus aureus, enterobacteria and C. albicans fungi, which indicates a violation of the microbiocenosis due to reduced mucosal immunity. Mucosal immunity of the oral cavity in children with mucoviscidosis is characterized by a 1.5-fold decrease in lysozyme activity and the level of secretory IgA in the saliva of children.

Respiratory viral infections drive different lung cytokine profiles in pigs.

BACKGROUND: Swine influenza A virus (IAV) and porcine reproductive and respiratory syndrome virus (PRRSV) are considered key viral pathogens involved in the porcine respiratory disease complex. Concerning the effect of one virus on another with respect to local immune response is still very limited. Determination of presence and quantity of cytokines in the lung tissue and its relation to the lung pathology can lead to a better understanding of the host inflammatory response and its influence on the lung pathology during single or multi-virus infection. The aim of the present study was to explore and compare the patterns of lung cytokine protein response in pigs after single or dual infection with swine IAV and/or PRRSV. RESULTS: Inoculation with IAV alone causes an increase in lung concentration of IFN-α, IFN-É£, TNF-α, IL-6, IL-8 and IL-10, especially at 2 and 4 DPI. In PRRSV group, beyond early IFN-α, IFN-É£, IL-6, IL-8 and IL-10 induction, elevated levels of cytokines at 10 and 21 DPI have been found. In IAV+PRRSV inoculated pigs the lung concentrations of all cytokines were higher than in control pigs. CONCLUSIONS: Current results indicate that experimental infection of pigs with IAV or PRRSV alone and co-infection with both pathogens induce different kinetics of local cytokine response. Due to strong positive correlation between local TNF-α and IL-10 concentration and lung pathology, we hypothesize that these cytokines are involved in the induction of lung lesions during investigates infection. Nevertheless, no apparent increase in lung cytokine response was seen in pigs co-inoculated simultaneously with both pathogens compared to single inoculated groups. It may also explain no significant effect of co-infection on the lung pathology and pathogen load, compared to single infections. Strong correlation between local concentration of TNF-α, IFN-É£, IL-8 and SwH1N1 load in the lung, as well as TNF-α, IL-8 and PRRSV lung titres suggested that local replication of both viruses also influenced the local cytokine response during infection.

Local immunity and microflora of the reproductive tract of gynecological patients before and after immunomodulatory therapy.

Immunomodulatory drugs are widely used as drugs for the treatment of inflammatory diseases of microbial etiology in clinical medicine. The authors evaluated the effectiveness of the therapeutic effect of the Cycloferon immunomodulator in the treatment of chronic purulent-inflammatory diseases of the reproductive tract using clinical, microbiological and immunological parameters. It was shown that under the influence of immunomodulatory therapy, the restoration of the qualitative and quantitative composition of the normoflora was observed with the elimination of etiologically significant microorganisms. The immunocorrective effect of therapy on the indices of local immunity was established: the concentration of lactoferrin increased in the cervical secretion, the level of cytokines IL-1ß and γ-IFN normalized, the amount of secretory IgA increased significantly, which contributed to the enhancement of local protective reactions, as well as the clinical efficacy of therapy, which was manifested in the reduction and/or disappearance of pain syndrome and the absence of relapse for 2 or more years. Conducted researches allow us to speak of immunotropic drugs as a promising direction in the treatment of inflammatory processes of the reproductive tract, a significant advantage of which is rational immunomodulation directly in the focus of inflammation, the availability of treatment, the absence of side effects with adequate therapeutic regimens, which makes it relevant to further study this direction of immunocorrection and its implementation into wide clinical practice.

Age-Associated Role of Lingual and Pharyngeal Glands in Local Immunity in Oral Cavity.

The macroscopic and histological methods were employed to examine the autopsy specimens of salivary lingual glands obtained from 299 patients of both sexes and various age ranging from newborn to longevity. The age-associated alterations of minor lingual and pharyngeal glands were revealed, and the topographical relations between the glands and lymphoid cells were described. The characteristic sparsity of the glands in infancy is caused by nutritional uniformity at this period, when diminished production of secretory IgA results in frequent inflammatory processes in oral and pharyngeal cavities. With age, the glandular orifices widen, and their number increases thereby augmenting local immunity in the oral cavity and in oral aspect of the pharynx. Starting from elderly and senile age, the involutive alterations were observed, which were accompanied by diminished production of secretory immunoglobulin A and related degradation of local and humoral immunity.

Use of technomolecular silver preparations in complex treatment of infected wounds.

OBJECTIVE: The aim: To improve the results of treatment of infected wounds. PATIENTS AND METHODS: Materials and methods:The clinical material is based on clinical observation and treatment of 29 patients with in fected wounds, whose treatment included combination drugs of local action on the basis of techno-molecular silver (in particular «Cadefort-Spray¼), by application to the wound surface. RESULTS: Results: Wound microbial factor, dynamics of wound process, indicators of immune status were evaluated: localadaptive immunity, atopic reactions. CONCLUSION: Conclusions: High efficiency of treatment was observed regardless of the phase of the wound process, which allowed to accelerate wound repair and stimulate the processes of regeneration, strengthen local adaptive immunity, prevent atopic reactions.

Vaccine-induced gastric CD4+ tissue-resident memory T cells proliferate in situ to amplify immune response against Helicobacter pylori insult.

BACKGROUND: Tissue-resident memory T cells accelerate the clearance of pathogens during recall response. However, whether CD4+ TRM cells themselves can provide gastric immunity is unclear. MATERIALS AND METHODS: We established a parabiosis model between the enhanced green fluorescent protein and wild-type mice that the circulation system was shared, and the wild-type partner was vaccinated with H pylori vaccine composed of CCF and silk fibroin in gastric subserous layer to induce gastric EGFP+ CD4+ TRM cells. Antigen-specific EGFP+ CD4+ T cells and proliferous TRM cells were analyzed by flow cytometry. The colonization of H pylori was detected by quantitative real-time PCR. EGFP+ CD4+ TRM cells and the inflammation of the stomach were observed by histology. RESULTS: A parabiosis animal model was employed to identify the cells that introduced by vaccination in GSL. Antigen-specific EGFP+ CD4+ T cells could be detected at day 7 post-vaccination. Thirty days later, EGFP+ CD4+ TRM cells were established with a phenotype of CD69+ CD103- . Of note, we found that when circulating lymphocytes were depleted by FTY720 administration, these TRM cells could proliferate in situ and differentiate into effector Th1 cells after H pylori challenge. A decrease in H pylori colonization was observed in the vaccinated mice but not unvaccinated mice. Further, we found that although FTY720 was administrated, mounted pro-inflammatory myeloid cells still emerged in the stomach of the vaccinated mice, which might contribute to the reduction of H pylori colonization. CONCLUSIONS: Our study reveals that H pylori vaccine-induced CD4+ TRM cells can proliferate and differentiate in situ to enhance gastric local immunity during recall response.

Local immunity status of oropharynx in patients with scleroma

Introduction: Scleroma is a rare chronic granulomatous disease of the upper respiratory tract caused by Klebsiella pneumoniae subsp. rhinoscleromatis. To date its pathogenesis is as yet little understood. At the same time, scleroma is associated with a number of immune system disturbances. The aim: To study local immunity status of oropharynx in patients with scleroma, and to compare its parameters in various clinical forms of the disease. Material and methods: 20 apparently healthy subjects and 92 patients with scleroma (33 males, 59 females) underwent clinical immunologic evaluation. There were 31 patients with dominating infiltrative form of scleroma, 30 ­ with dominating atrophic form, 31 ­ with dominating scarring form. Concentration of secretory and monomeric immunoglobulin A, immunoglobulin G, α-interferon, interleukin 1ß in oropharyngeal secretion was determined by enzyme immunoassay. Resluts: Patients with scleroma were found to have altered local immunity of oropharyngeal secretion. There was a strong tendency for decreased concentration of secretory immunoglobulin A - 1.3-2.0 times, and decreased immunoglobulin G level ­ 1.5-2.3 times (р < 0.05) as compared to the values in healthy subjects. Specific features of local immunity in oropharyngeal secretion in various forms of scleromatous inflammatory process in upper respiratory tract were found: the most significant decrease of α-interferon concentration in atrophic and scarring forms of the disease, and the largest increase of anti-inflammatory interleukin 1ß and immune complex concentration in infiltrative form of scleroma. Conslusions: The study revealed deficiency of local immunity factors in oropharynx, being indicative of immunopathogenetic role of diagnosed disturbances in development and persistence of chronic inflammation in scleroma, and emphasizing the necessity of immunocorrection in complex therapy of the disease.

[Features of local immunity in local inflammatory reactions in pregnant, depending on the implementation of intrauterine infection].

OBJECTIVE: Introduction: Vaginal protective systems function independently of other systemic mechanisms and impairments of local immune mechanisms of the vaginal mucosa are the cause of infectious disorders of the reproductive system The aim: To assess characteristics of local immunity in local inflammatory reactions in pregnant, depending on the implementation of intrauterine infection. PATIENTS AND METHODS: Materials and methods: The study comprised Group 1 of pregnant women with the presence of bacterial infections with and without implementation of IUI, Group 2 with the presence of viral infections with and without implementation of IUI, Group 3 with the presence of infections of combined polyetiological structure with and without implementation of IUI and control group included pregnant women with physiological pregnancy. The study implied evaluation of the number of leukocytes in vaginal contents, concentration of IgA, IgM, IgG, sIgA, concentration of cytokines-IL-1 ß , IL-6, IL-10, TNF- α in vaginal swabs. RESULTS: Results: Women with the presence of an infectious process without its implementation, regardless of the etiologic factor, were found to have more expressed inflammatory reactions. In the presence of a viral infection, changes in the humoral link of local immunity were less expressed. In Group 1 without implementation of IUI, the concentration of IL-1 ß was (73.68 ± 10.23) pg / ml, IL-6 was (53.3 ± 6.8) pg / ml. In Group 2 with implementation of IUI, concentration of IL-10 was (26.72 ± 4.35) pg / ml. In Group 2 without implementation of IUI, the content of TNF-α was (575.25 ± 69.03) pg / ml. CONCLUSION: Conclusions: The study showed the differences between the groups of pregnant women with different outcome of IUI in the content of proinflammatory and anti-inflammatory cytokines.

[Clinical and immunological features of oral and vermillion border precancer diseases]. / Kliniko-immunologicheskie osobennosti predrakovykh zabolevanii slizistoi obolochki rta i krasnoi kaimy gub.

Research objective was assessment of a possibility of primary diagnosis of viruses Epstein-Burr, a cytomegalovirus, a virus of herpes of the 6th type in oral liquid and also influences of herpes infection on development and the clinical course of precancer diseases of the oral mucosa and the red border of lips (RBL). Profound clinic-immunological examination is conducted to 60 patients: the first group are have made 20 patients with an erosive and ulcer form of the lichen planus, the second - 20 people with an erosive form of a leukoplakia, a third - 20 people without diseases of the oral mucosa. As a result of work detection herpes infection is revealed high (90%). The combined infection with a virus Epstein-Burr and a virus of herpes of the 6th type was observed more than at a half of patients. The imbalance of factors of local immunity of the oral cavity in the form of increase in the IgG profile, decrease in concentration of IGA, lysozyme, and increase in an indicator of Ksb three times in comparison with norm and also substantial increase of level of pro-inflammatory cytokin IL 1ß and FNO-α is found in patients with precancer diseases of the oral mucosa. Characteristic clinical feature of the precancer diseases associated with latent herpes infection is the long, recidivous current, persistent to traditional therapy. The research of oral liquid and blood on herpes viruses and consultation of the infectiologist is recommended to all patients with precancer diseases.

[The development of diagnostic of acute endometritis on the basis of modeling and cluster analysis of parameters of local immunity.]

The acute endometritis provoking expressed disorders of local immunity is considered as one of the prevalent complications of postnatal period and abortions. The purpose of study is to develop diagnostic of acute endometritis on the basis of applied correlation modeling and cluster analysis techniques diagnostically priority-driven immunological indices of vaginal cervical mucus. The sample included main group (154 patients with acute endometritis) and control group (103 patients with uncomplicated post-natal period). Both groups were subjected to enzyme-linked immunosorbent assay to analyze content of immunoglobulins, components of complement and lysozyme in vaginal cervical mucus. The sampling of diagnostically significant indices was implemented using correlation and cluster analysis techniques. It is established that chief diagnostic indices of local immunity in case of acute endometritis are IG M, sIg A, С4 components of complement and lysozyme. The established diagnostically significant parameters of local immunity provide development of detection of acute endometritis according less number of indices.

Transcriptomic variation of locally-infected skin of Epinephelus coioides reveals the mucosal immune mechanism against Cryptocaryon irritans.

Fish skin is the largest immunologically active mucosal organ, providing first-line defense against external pathogens. However, the skin-associated immune mechanisms of fish are still unclear. Cryptocaryon irritans is an obligate ectoparasitic ciliated protozoan that infects almost all marine fish, and is believed to be an excellent pathogen model to study fish mucosal immunity. In this study, a de novo transcriptome assembly of Epinephelus coioides skin post C. irritans tail-infection was performed for the first time using the Illumina HiSeq™ 2500 system. Comparative analyses of infected skin (group Isk) and uninfected skin (group Nsk) from the same challenged fish and control skin (group C) from uninfected control fish were conducted. As a result, a total of 91,082 unigenes with an average length of 2880 base pairs were obtained and among them, 38,704 and 48,617 unigenes were annotated based on homology with matches in the non-redundant and zebrafish database, respectively. Pairwise comparison resulted in 10,115 differentially-expressed genes (DEGs) in the Isk/C group comparison (4,983 up-regulated and 5,132 down-regulated), 2,275 DEGs in the Isk/Nsk group comparison (1,319 up-regulated and 956 down-regulated) and 4,566 DEGs in the Nsk/C group comparison (1,534 up-regulated and 3,032 down-regulated). Seven immune-related categories including 91 differentially-expressed immune genes (86 up-regulated and 5 down-regulated) were scrutinized. Both DEGs and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and immune-related gene expression analysis were used, and both analyses showed that the genes were more significantly altered in the locally-infected skin than in the uninfected skin of the same challenged fish. This suggests the skin's local immune response is important for host defense against this ectoparasite infection. Innate immune molecules, including hepcidin, C-type lectin, transferrin, transferrin receptor protein, serum amyloid A, cathepsin and complement components were significantly up-regulated (fold-change ranged from 3.3 to 12,944) in infected skin compared with control skin. The up-regulation of chemokines and chemokine receptors and activation of the leukocyte transendothelial migration pathway suggested that leucocytes intensively migrated to the local infected sites to mount a local immune defense. Toll-like receptors (TLRs) 1, 2, 5 and 5S were most significantly up-regulated in the infected skin, suggesting that these TLRs may be involved in parasite pathogen-associated molecular pattern (PAMPs) recognition. Up-regulation of the dendritic cell markers CD209 and CD83 and other antigen presentation pathway molecules provided evidence for skin local antigen presentation. Up-regulation of the T cell markers CD4 and CD48, B cell markers CD22 and CD81 and B cell receptor signaling kinase Lyn, showed the presence and population expansion of T/B cells at locally-infected sites, which suggested possible activation of a local specific immune response in the skin. Our results will facilitate in-depth understanding of local immune defense mechanisms in fish skin against ectoparasite infection.

Persistence and evolution of allergen-specific IgE repertoires during subcutaneous specific immunotherapy.

BACKGROUND: Specific immunotherapy (SIT) is the only treatment with proved long-term curative potential in patients with allergic disease. Allergen-specific IgE is the causative agent of allergic disease, and antibodies contribute to SIT, but the effects of SIT on aeroallergen-specific B-cell repertoires are not well understood. OBJECTIVE: We sought to characterize the IgE sequences expressed by allergen-specific B cells and track the fate of these B-cell clones during SIT. METHODS: We used high-throughput antibody gene sequencing and identification of allergen-specific IgE with combinatorial antibody fragment library technology to analyze immunoglobulin repertoires of blood and the nasal mucosa from aeroallergen-sensitized subjects before and during the first year of subcutaneous SIT. RESULTS: Of 52 distinct allergen-specific IgE heavy chains from 8 allergic donors, 37 were also detected by using high-throughput antibody gene sequencing of blood samples, nasal mucosal samples, or both. The allergen-specific clones had increased persistence, higher likelihood of belonging to clones expressing other switched isotypes, and possibly larger clone size than the rest of the IgE repertoire. Clone members in nasal tissue showed close mutational relationships. CONCLUSION: In the future, combining functional binding studies, deep antibody repertoire sequencing, and information on clinical outcomes in larger studies might aid assessment of SIT mechanisms and efficacy.

Influence of medium-chain fatty acids and short-chain organic acids on jejunal morphology and intra-epithelial immune cells in weaned piglets.

Medium-chain fatty acids (MCFA) and short-chain organic acids (SOA) are often used as feed additives in piglet diets. There are limited studies in pigs describing the impact of MCFA or SOA on gut morphology and the local immune system. The aim of this study was to investigate whether the supplementation of SOA (0.41% fumaric acid and 0.32% lactic acid), or the combination of SOA with MCFA (0.15% caprylic and capric acid) would have effects on gut morphology and intestinal immune cells in weaned piglets. A total number of 72 weaned piglets were randomly allocated into three experimental groups. Tissue samples of six animals per group were used to investigate the potential impact of the feed additives on villus length and crypt depth of the jejunum and to quantify intra-epithelial lymphocytes (IEL). CD3-positive IEL were determined via immunohistochemistry (IHC) and flow cytometry (FC), whereas CD2-, CD5-, CD8ß-, CD16- and γδ TCR-positive IEL were only analysed by FC. The supplementation of MCFA and SOA did not significantly affect morphometric data. The FC data indicated that SOA significantly increased the quantity of CD2- CD8- γδ T cells in the jejunum epithelium. Both IHC and FC analyses of pig jejunum confirmed that the majority of IEL expressed the surface marker CD3 and could be classified as cytotoxic T lymphocytes. In conclusion, the data indicated that SOA increased the proportion of CD2- CD8- γδ T cells in the jejunal epithelium. Thus, SOA might enable a beneficial effect on the local immunity by increasing the constitutive number of potential effector cells to defeat infectious diseases.

Influence of oral application of Enterococcus faecium AL41 on TGF-ß4 and IL-17 expression and immunocompetent cell distribution in chickens challenged with Campylobacter jejuni.

Campylobacteriosis is mainly caused by infection with Campylobacter jejuni following consumption or handling of Campylobacter-contaminated poultry meat. The aim of this study was to investigate the effect of probiotic Enterococcus faecium AL41 on TGF-ß4 and IL-17 expression and on immunocompetent cell distribution after C. jejuni infection in broiler chicken, as a second part of the previous study of Karaffová et al. (2017). Accordingly, day-old chicks were randomly divided into four experimental groups of 10 chicks each (n = 10): control (C), E. faecium AL41 (EFAL41), C. jejuni CCM6191 (CJ), and combined E. faecium AL41 + C. jejuni CCM6191 (EFAL41 + CJ). Samples from the caecum were collected on days 4 and 7 post Campylobacter infection (dpi), for the isolation of mRNA of TGF-ß4, IL-17 and for immunohistochemistry. The relative mRNA expression of TGF-ß4 was upregulated in the combined (EFAL41 + CJ) group compared to other groups during both samplings, but the expression of IL-17 was downregulated. Similarly, the highest density of CD3+ was detected in the combined group at 7 dpi, but the number of IgA+ cells was increased in both groups with EFAL41. It was concluded that the EFAL41 probiotic E. faecium strain can modulate the expression of selected cytokines (upregulation of TGF-ß4 but downregulation of IL-17 relative expression), and activate IgA-producing cells in the caeca of chicks infected with C. jejuni CCM6191.

Proteflazid® and local immunity in diseases caused by human papillomavirus, herpesvirus and mixed urogenital infections.

Reporting of clinical trials results for Proteflazid® in the drug formulation suppositories and vaginal swabs soaked in the solution of the drug to the local immunity of the female reproductive tract. AIM: The aim of study was to examine the state of local immunity in the reproductive tract of women with sexually transmitted diseases caused by human papillomavirus, herpes viruses (Type 1, 2) and mixed infection (herpes viruses + chlamydia). MATERIALS AND METHODS: The trials involved 216 women with viral sexually transmitted diseases: Cervical Dysplasia associated with papillomavirus infection (HPV) (Group 1); Herpes genitalis type 1 (HSV- 1) and type 2 (HSV-1) (Group 2); mixed infection - HSV-1, HSV-2 and chlamydia (Group 3). RESULTS: Treatment results have confirmed that Proteflazid® contributes to sustainable performance improvement of basic factors of local immunity - sIgA, lysozyme and complement component C3 in the cervical mucus for all three groups of women. CONCLUSIONS: Proteflazid® enhances level of local immunity markers (sIgA, lysozyme, C3 complement component) and improves their ratios. Also it intensifies anticontagious activity of mucosal protection and female reproductive system as whole, during treatment diseases caused by human papillomavirus, herpesvirus and mixed urogenital infections (herpesvirus and chlamydia).