Secretory IgA Levels Research Articles

Virulence Associated Genes-Deleted Salmonella Montevideo Is Attenuated, Highly Immunogenic and Confers Protection against Virulent Challenge in Chickens.

To construct a novel live vaccine against Salmonella enterica serovar Montevideo (SM) infection in chickens, two important bacterial regulatory genes, lon and cpxR, which are associated with invasion and virulence, were deleted from the wild type SM genome. Attenuated strains, JOL1625 (Δlon), JOL1597 (ΔcpxR), and JOL1599 (ΔlonΔcpxR) were thereby generated. Observations with scanning electron microscopy suggested that JOL1625 and JOL1599 cells showed increased ruffled surface which may be related to abundant extracellular polysaccharide (EPS) production. JOL1597 depicted milder ruffled surface but showed increased surface corrugation. ConA affinity-based fluorometric quantification and fluorescence microscopy revealed significant increases in EPS production in JOL1625 and JOL1599. Four weeks old chickens were used for safety and immunological studies. The mutants were not observed in feces beyond day 3 nor in spleen and cecum beyond day 7, whereas wild type SM was detected for at least 2 weeks in spleen and cecum. JOL1599 was further evaluated as a vaccine candidate. Chickens immunized with JOL1599 showed strong humoral responses, as indicated by systemic IgG and secretory IgA levels, as well as strong cell-mediated immune response, as indicated by increased lymphocyte proliferation. JOL1599-immunized groups also showed significant degree of protection against wild type challenge. Our results indicate that Δlon- and/or ΔcpxR-deleted SM exhibited EPS-enhanced immunogenicity and attenuation via reduced bacterial cell intracellular replication, conferred increased protection, and possess safety qualities favorable for effective vaccine development against virulent SM infections.

Intranasal immunization with recombinant Toxoplasma gondii actin depolymerizing factor confers protective efficacy against toxoplasmosis in mice.

Background Toxoplasma gondii is an opportunistic protozoan closely associated with AIDS and vertical transmission. T. gondii actin depolymerizing factor (TgADF) plays an important role in actin cytoskeleton remodeling, and it is required to invade host cells. TgADF was a promising vaccine candidate. To observe the immunological changes and protective efficacy of recombinant TgADF protein (rTgADF) against T. gondii infection, we optimized the intranasal immunization dose of rTgADF and analyzed the survival rate and tachyzoite loads in mouse tissues after oral challenge with T. gondii tachyzoites.ResultsrTgADF was prepared, purified, and combined with mouse anti-His antibody and rabbit anti-T. gondii serum. After intranasal immunization with 10 μg, 20 μg, 30 μg, or 40 μg of rTgADF, the 30-μg group elicited high levels of secretory IgA (sIgA) in nasal, intestinal, and vesical washes, raised IgG titres in the sera, strong proliferation of splenocytes, and increased secretion of IL-2 and IFN-γ when compared with the control group. When the mice were orally challenged with T. gondii, an increase in the survival rate (36.36 %) and a decrease in the tachyzoite loads in the liver (67.77 %) and brain (51.01 %) were observed.ConclusionsOur findings demonstrate that intranasal immunization with rTgADF can simultaneously trigger mucosal and systemic immune responses and protect the mice against T. gondii infection.

Secretory immunoglobulin A and immunoglobulin G in horse saliva

This study aimed to increase the knowledge on salivary antibodies in the horse since these constitute an important part of the immune defence of the oral cavity. For that purpose assays to detect horse immunoglobulin A (IgA) including secretory IgA (SIgA) were set up and the molecular weights of different components of the horse IgA system were estimated. Moreover, samples from 51 clinically healthy horses were tested for total SIgA and IgG amounts in saliva and relative IgG3/5 (IgG(T)) and IgG4/7 (IgGb) content were tested in serum and saliva.Results showed a mean concentration of 74μg SIgA/ml horse saliva and that there was a large inter-individual variation in salivary SIgA concentration. For total IgG the mean concentration was approx. 5 times lower than that of SIgA, i.e. 20μg IgG/ml saliva and the inter-individual variation was lower than that observed for SIgA. The saliva–serum ratio for IgG isotypes IgG3/5 and IgG4/7 was also assessed in the sampled horses and this analysis showed that the saliva–serum ratio of IgG4/7 was in general approximately 4 times higher than that of IgG3/5.The large inter-individual variation in salivary SIgA levels observed for the normal healthy horses in the present study emphasises the need for a large number of observations when studying this parameter especially in a clinical setting. Moreover, our results also indicated that some of the salivary IgG does not originate from serum but may be produced locally. Thus, these results provide novel insight, and a base for further research, into salivary antibody responses of horses.

Effects of lipopolysaccharide on the histomorphology and expression of toll-like receptor 4 in the chicken trachea and lung

ABSTRACTEndotoxin or lipopolysaccharide (LPS) exposure can cause injury to the respiratory airways and in response, the respiratory epithelia express toll-like receptors (TLRs) in many species. However, its role in the innate immunity in the avian respiratory system is poorly understood. The aim of the present study was to evaluate the effects of LPS on the chicken trachea and lung. After intraperitoneal LPS or saline injection, the trachea and lungs were harvested at 0, 12, 36 and 72 h (n = 6 at each time point) and histopathologically analysed using haematoxylin and eosin and periodic acid-Schiff staining, while TLR4 expression was determined by immunohistochemistry and secretory Immunoglobulin A (SIgA) levels by enzyme-linked immunosorbent assay. After LPS stimulation, we observed a remarkable decrease in the number of goblet cells along with obvious disruption and desquamation of the ciliated epithelium in the trachea, blurring of the boundary between pulmonary lobules, narrowed or indistinguishable lumen of the pulmonary atria and leukostasis in the lungs. Following LPS stimulation, TLR4 protein expression was up-regulated in both the trachea and the lungs and was found on the ciliated columnar cells as well as in the submucosa of the trachea, and in the lungs on parenchymal and immune cells. However, SIgA levels were only up-regulated in the trachea at 12 h following LPS stimulation. Hence, this report provides novel information about the effects of LPS on the microstructure of the lower respiratory tract and it is concluded that its intra-peritoneal administration leads to TLR4-mediated destruction of the tracheal epithelium and pulmonary inflammation along with increased SIgA expression in the tracheal mucosa.

Abstract # 1850 Early life stress predisposes to increased sensitivity to inflammatory stimuli through intestinal dysbiosis

Stress, anxiety and depression are considered risk factors for Inflammatory Bowel Disease. We have previously shown in mice that early life stress induced by maternal separation (MS) changes host physiology leading to intestinal dysbiosis, which in turns causes anxiety and depression-like behavior. Using this murine model, we investigated the role of stress and microbiota in susceptibility to intestinal inflammation. Adult MS and control C57Bl/6 mice were used. Experimental colitis was induced by 1.5% or 3.5% dextran sulfate sodium (DSS) in drinking water in germ-free or colonized (ex-germ-free) mice, respectively. After their behavior and clinical scores were assessed, mice were sacrificed and samples collected. Colonized but not germ-free MS mice displayed anxiety and depression-like behavior, higher secretory IgA levels, decreased Paneth cell counts and dysbiosis compared to controls. In colonized mice, DSS colitis clinical scores were higher in MS mice accompanied by marked dysbiosis, lower secretory IgA and increased serum MCP-1 levels compared to controls, although the colonic microscopic scores and inflammatory cytokine levels were similar. In germ-free conditions, no difference in clinical colitis scores, histology or cytokine levels were found. In summary, MS leads to intestinal dysbiosis that sustains and exacerbates an impaired cellular and humoral immunity, leading to increased sensitivity to intestinal injury. Our results further support the role of gut-microbiota-brain axis in the genesis of chronic gastrointestinal inflammatory disorders.

Potential of Translationally Controlled Tumor Protein-Derived Protein Transduction Domains as Antigen Carriers for Nasal Vaccine Delivery.

Nasal vaccination offers a promising alternative to intramuscular (i.m.) vaccination because it can induce both mucosal and systemic immunity. However, its major drawback is poor absorption of large antigens in the nasal epithelium. Protein transduction domains (PTDs), also called cell-penetrating peptides, have been proposed as vehicles for nasal delivery of therapeutic peptides and proteins. Here, we evaluated the potential of a mutant PTD derived from translationally controlled tumor protein (designated TCTP-PTD 13) as an antigen carrier for nasal vaccines. We first compared the l- and d-forms of TCTP-PTD 13 isomers (l- or d-TCTP-PTD 13) as antigen carriers. Studies in mice demonstrated that nasally administered mixtures of the model antigen ovalbumin (OVA) and d-TCTP-PTD 13 induced higher plasma IgG titers and secretory IgA levels in nasal washes than nasally administered OVA alone, OVA/l-TCTP-PTD 13, or i.m.-injected OVA. Plasma IgG subclass responses (IgG1 and IgG2a) of mice nasally administered OVA/d-TCTP-PTD 13 showed that the predominant IgG subclass was IgG1, indicating a Th2-biased immune response. We also used synthetic CpG oligonucleotides (CpG) as a Th1 immune response-inducing adjuvant. Nasally administered CpG plus OVA/d-TCTP-PTD 13 was superior in eliciting systemic and mucosal immune responses compared to those induced by nasally administered OVA/d-TCTP-PTD 13. Furthermore, the OVA/CpG/d-TCTP-PTD 13 combination skewed IgG1 and IgG2a profiles of humoral immune responses toward a Th1 profile. These findings suggest that TCTP-derived PTD is a suitable vehicle to efficiently carry antigens and to induce more powerful antigen-specific immune responses and a more balanced Th1/Th2 response when combined with a DNA adjuvant.

ON THE ASSESSMENT AND TREATMENT OF PATIENTS WITH CHRONIC PROSTATITIS IN PERSISTENT UROGENITAL CHLAMYDIOSIS

Актуальность. Урогенитальный хламидиоз – наиболее распространенная бактериальная инфекция, передаваемая половым путем, сопровождающаяся высокой частотой осложнений (хронический простатит, сальпингоофорит и др.) и неблагоприятных последствий (нарушение половой и репродуктивной функций). В последние годы отмечается нарастание персистирующей хламидийной инфекции, которая не поддается лечению антибиотиками и диагностируется на основании обнаружения методом иммуноферментного анализа антител иммуноглобулина (Ig) класса G к белку теплового шока хламидий с молекулярной массой 60 кД (HSP60) и определения секреторного IgA к Chlamydia trachomatis в семенной плазме. Цель – выявление частоты пер- систирующей хламидийной инфекции у больных хроническим простатитом путем определения сывороточных антител IgG к HSP60 C. trachomatis и наличия противохламидийных IgA в семенной плазме. Материал и методы. В рамках проспективного исследования мы провели обследование 18 больных хроническим простатитом в возрасте от 21 до 58 лет, в контрольную группу вошли 30 человек без хламидийной урогенитальной патологии. Клиническое обследование включало сбор анамнеза, инструментальное обследование, ультразвуковое исследование предстательной железы и семенных пузырьков. Серологические исследования проводились с помощью иммуноферментного анализа в четырех тест-системах фирмы “Medac” (Германия) к двум антигенам: родоспецифическому липополисахариду хламидий r-ELISA (антитела классов IgM, IgG и IgA к Chlamydia) и основному белку наружной мембраны (МОМР) C. trachomatis (анти- тела классов IgG и IgA). Антитела к HSP60 хлами- дий определяли в тест-системе c HSP60-IgG-ELISA фирмы “Medac”. Определение секреторного IgA в семенной жидкости проводили с помощью тест-системы Chlamydia trachomatis-IgA-pELISA. Результаты. В основной группе в 13 (72,2%) из 18 случаев выявлен хронический хламидийный простатит, который в 7 (38,9%) случаях протекал на фоне персистирующей хламидийной инфекции, сопровождаясь одновременным повышением титра сывороточных антител IgG к HSP60 C. trachomatis и наличием противохламидийных антител IgA в семенной плазме. У 30 че- ловек контрольной группы сывороточные антитела IgG к C. trachomatis обнаружены не были. Заключение. Полученные данные подтверждают диагностическую ценность определения присутствия IgA к С. trachomatis с целью установления этиологии воспалительного процесса в мужском генитальном тракте, а также свидетельствуют о высокой частоте персистирующей хламидийной инфекции у больных хроническим простатитом и целесообразности использования серологического обследования на наличие сывороточных антител IgG к HSP60 C. trachomatis и противохламидийных IgA в семенной плазме.

Relationship between salivary stress biomarker levels and cigarette smoking in healthy young adults: an exploratory analysis.

BackgroundThis study investigated the relationships among salivary stress biomarkers, cigarette smoking, and mood states.MethodsThe study population comprised 49 healthy sixth-year dental students at Fukuoka Dental College (39 men, 10 women; age, 23–31 years). Lifetime exposure to smoking was calculated using the Brinkman index (BI). Resting saliva samples were collected, and concentrations of cortisol, secretory immunoglobulin A (SIgA), interleukin (IL)-1β, interleukin-6, and tumor necrosis factor (TNF)-α were determined. Mood states (tension-anxiety, depression-dejection, anger-hostility, fatigue, confusion, and vigor) over the previous week were assessed using the Profile of Mood States - Brief Japanese Version.ResultsSalivary IL-1β levels were significantly higher in smokers than non-smokers (P = 0.044), regardless of the BI or mood state. A significant positive correlation was evident between the TNF-α level and the BI (P = 0.036), and the SIgA level was positively correlated with the BI (P = 0.067) but did not reach statistical significance. In terms of mood states, higher fatigue scores and lower vigor scores were observed in smokers. The TNF-α level and vigor score were negatively correlated (r = –0.229, P = 0.135), but the correlation did not reach statistical significance. However, the SIgA level and fatigue score were significantly positively correlated (r = 0.410, P = 0.005).ConclusionsThe TNF-α and SIgA levels were both positively correlated with the BI. Furthermore, the TNF-α level was negatively correlated with the vigor score, whereas the SIgA level was positively correlated with the fatigue score. Thus, salivary levels of TNF-α and SIgA may be used as biomarkers of mood states in healthy young smokers.

Clinical and Immunological Benefits of OM-85 Bacterial Lysate in Patients with Allergic Rhinitis, Asthma, and COPD and Recurrent Respiratory Infections.

PurposeThe aim of this study was to evaluate the efficacy of OM-85 in reducing the incidence of respiratory tract infections (RTIs) in patients with allergic rhinitis, asthma, or chronic obstructive pulmonary disease (COPD), and its effect on immunological parameters, namely serum and secretory IgA levels.MethodsThis was an open-label, prospective, sequential study which included 84 consecutive patients aged 16–65 years, who presented with recurrent (three or more) respiratory infections during the year prior to study entry. In the first year of the study, patients received standard optimized care (SOC), according to their underlying disease condition (asthma, allergic rhinitis, or COPD). In the following year, patients received treatment with OM-85 oral bacterial lysate (one 7 mg capsule daily for ten consecutive days per month, for 3 months), with a 6-month follow-up. Medical history, clinical symptoms, serum, and secretory IgA levels, and the number of infections and exacerbations were evaluated before and after treatment.ResultsThere was a decrease in the total number of RTIs before the OM-85 treatment period (SOC only) compared to the year before the study start [69/266 (corresponding to a 74 % reduction)] and an additional decrease [38/69 (corresponding to a 45 % reduction)] after OM-85 treatment; p < 0.05. There was also a significant reduction in the total number of exacerbations related to the patients’ underlying medical conditions, which decreased from 55 to 35 during OM-85 (+SOC) treatment, corresponding to a reduction of 36 %. In addition, an increase in serum and secretory IgA levels which coincided with the administration of OM-85 was observed.ConclusionsOur results showed the clinical benefits of OM-85 in reducing RTIs and exacerbations of the underlying medical condition, in patients with allergic rhinitis, asthma, or COPD.

Lactobacillus reuteri supplements do not affect salivary IgA or cytokine levels in healthy subjects: A randomized, double-blind, placebo-controlled, cross-over trial

Objectives: To evaluate the effect of daily ingestion of probiotic lactobacilli on the levels of secretory IgA (sIgA) and selected cytokines in whole saliva of healthy young adults.Materials and methods: The study group consisted of 47 healthy adults (18–32 years) who volunteered for a randomized, double-blind, placebo-controlled, cross-over trial after informed consent. During intervention, the subjects ingested two lozenges per day containing two strains of the probiotic bacterium Lactobacillus reuteri (DSM 17938 and ATCC PTA 5289) or placebo lozenges. The intervention and wash-out periods were 3 weeks. Saliva samples were collected at baseline, immediately after each intervention period and 3 weeks post-intervention. ELISA was used to measure sIgA and luminex technology was used to measure the interleukins (IL)-1β, IL-6, IL-8 and IL-10. For statistical analyses a mixed ANOVA model was employed to calculate changes in the salivary outcome variables.Results: Forty-one subjects completed the study and reported a good compliance. No significant differences in the concentrations of salivary sIgA or cytokines were recorded between the L. reuteri and placebo interventions or between baseline and 3 weeks post-intervention levels. No side- or adverse effects were reported.Conclusions: Supplementation with two strains of the probiotic L. reuteri did not affect sIgA or cytokine levels in whole saliva in healthy young adults. The results thereby indicate that daily oral supplementation with L. reuteri do not seem to modulate the salivary oral immune response in healthy young subjects (ClinicalTrials.gov NCT02017886).

Effects of Probiotics on Intestinal Mucosa Barrier in Patients With Colorectal Cancer after Operation: Meta-Analysis of Randomized Controlled Trials.

Many studies have found that probiotics or synbiotics can be used in patients with diarrhea or inflammatory bowel disease for the prevention and treatment of some pathologies by improving gastrointestinal barrier function. However, there are few studies availing the use of probiotics in patients with colorectal cancer. To lay the foundation for the study of nutritional support in colorectal cancer patients, a meta-analysis has been carried out to assess the efficacy of probiotics on the intestinal mucosa barrier in patients with colorectal cancer after operation.To estimate the efficacy of probiotics on the intestinal mucosa barrier in patients with colorectal cancer after operation, a meta-analysis of randomized controlled trials has been conducted.Databases including PubMed, Ovid, Embase, the Cochrane Central Register of Controlled Trials, and the China National Knowledge Infrastructure have been searched to identify suitable studies. Stata 12.0 was used for statistical analysis, and sensitivity analysis was also conducted. Six indicators were chosen to evaluate probiotics in protecting the intestinal mucosa barrier in patients with colorectal cancer. Ratios of lactulose to mannitol (L/M) and Bifidobacterium to Escherichia (B/E), occludin, bacterial translocation, and levels of secretory immunoglobulin A (SIgA), interleukin-6 (IL-6), and C-reactive protein (CRP) were chosen to evaluate probiotics in protecting the intestinal mucosa barrier in patients with colorectal cancer.Seventeen studies including 1242 patients were selected for meta-analysis, including 5 English studies and 12 Chinese studies. Significant effects were found in ratios of L/M (standardized mean difference = 3.83, P = 0.001) and B/E (standardized mean difference = 3.91, P = 0.000), occludin (standardized mean difference = 4.74, P = 0.000), bacterial translocation (standardized mean difference = 3.12, P = 0.002), and levels of SIgA (standardized mean difference = 2.91, P = 0.004) and CRP (standardized mean difference = 4.21, P = 0.000), but no significant effects were found for levels of IL-6 (standardized mean difference = 1.33, P = 0.184).Probiotics can effectively protect the intestinal mucosa physical and biological barrier in patients with colorectal cancer after operation. However, to evaluate the protective effect on intestinal mucosal barrier, further studies on the type and concentration of the probiotics, duration of therapy, and the therapeutic route are required.

Twelve weeks of soccer-specific training: effects on mucosal immunity, salivary alpha-amylase and body composition in male African youths

The primary aim of this study was to determine the levels of salivary secretory IgA (sIgA) and salivary alpha-amylase (sAA) in young, black male soccer players, before and after 12 weeks of soccer-specific training. Thirty-four children (11–13 years) who were part of a youth soccer development training academy, participated in the study. The participants underwent 12 weeks of soccer-specific training. Resting saliva samples were collected 48 h before the commencement, and 48 h after the completion, of the training program. Samples were taken between 07:30 and 08:30, 90 min after waking. Body fat percentage (BF %), lean body mass (LBM) and cardiorespiratory fitness (CRF) were also measured. Significant differences were found between pre- and post-training for body mass index (BMI) (P 0.05), sAA secretion rate (P > 0.05) or sIgA concentration (P > 0.05). The magnitude of differences from pre- to post-training applying Cohen’s d effect sizes (ES) were moderate (>0.5) for estimated VO2max, sAA, sAA secretion rate, sIgA and sIgA secretion rate. These findings suggest that, 12 weeks of soccer-specific training enhances mucosal immunity and body composition and may have an effect on the sympathetic nervous system in black, male youths.

Salivary profile in diabetic patients: biochemical and immunological evaluation.

BackgroundThe aim of this study was to evaluate the biochemical and immunological characteristics of saliva from diabetic patients compared to non-diabetic adults.MethodsEighty-eight diabetic adults and 39 non-diabetic adults (control) were included in the study. Glucose, urea, calcium, total protein and amylase were determined by a colorimetric method. The levels of secretory IgA and the IgA anti-Streptococcus mutans and anti-insulin IgA antibodies were measured by enzyme-linked immunosorbent assay (ELISA). Caries status was evaluated using the DMFT index.ResultsGlucose, urea, calcium, anti-S. mutans IgA, total IgA, and anti-insulin IgA were significantly higher in diabetic patients, whereas total protein and amylase levels were lower in these patients. There was no positive correlation between blood and salivary glucose levels in either group. Diabetic patients had a higher DMFT index.ConclusionsThe present study showed for the first time that IgA levels in diabetic patients’saliva, shows correlation with systemic biochemical parameters. Thus the saliva is an useful tool to follow the systemic health status in these patients.

Intranasal Vaccination against HIV-1 with Adenoviral Vector-Based Nanocomplex Using Synthetic TLR-4 Agonist Peptide as Adjuvant.

Recombinant type 5 adenovirus (rAd5) vaccines hold the promise to prevent HIV-1 infections. Intranasal vaccination not only stimulates systemic immunity but also elicits mucosal immunity that provides first defense for mucosally transmitted diseases like HIV-1. Adjuvants such as TLR agonists are usually codelivered with antigens to enhance the immunogenicity of vaccines. Here, we present a rAd5 vaccine delivery system using DEG-PEI as the carrier. Adenovirus encoding HIV gag was used as antigen, and was complexed with DEG-PEI polymer via electrostatic interaction. A novel synthetic TLR-4 agonist, RS09, was either chemically linked with DEG-PEI (DP-RS09) or physically mixed with it(DP/RS09) to enhance the immunogenticity of rAd5 vaccine. After intranasal immunization, the systemic antigen-specific immune responses and cytotoxicity T lymphocytes responses induced by DP-RS09-rAd5 and DP/RS09-rAd5 were analyzed. The mucosal secretory IgA level was detected in both nasal and vaginal washes to determine the mucosal immunity. Furthermore, cytokine productions on RAW264.7 cells were tested after preincubation with TLR-4 pathway inhibitors. The results indicated that DEG-PEI could facilitate the intranasal delivery of rAd5 vaccine. Both chemically linked (DP-RS09) and physically mixed RS09 (DP/RS09) could further enhance the mucosal immunity of rAd5 vaccine via TLR-4 pathway. This RS09 adjuvanted DEG-PEI polymer represents a potential intranasal vaccine delivery system and may have a wider application for other viral vectors.

CORRECTION OF DYSBIOTIC ABNORMALITIES IN CHILDREN WITH ACUTE RESPIRATORY DISORDERS

Background: A special attention in the treatment of acute respiratory disorders (ARD) in children should be paid to correction of defense mechanisms of the body, including elimination of dysbiotic abnormalities. The use of probiotics, whose mechanism of action is directed to restoration of qualitative and quantitative composition of normal microbiota, is considered to be perspective in the combination therapy of ARD patients. Aim: to assess clinical and laboratory efficacy of probiotic Florin forte in children with ARD. Materials and methods: One hundred and eleven children aged from3 months to 14 years with ARD were included into the study. In 81.1% of cases they had concomitant obstruction of upper respiratory ways. From day1 after admission to the hospital, 81 patients (the main group) were administered probiotic Florin forte as a part of combination therapy for 5 to7 days, and 30 children (the comparator group) were administered the standard treatment without probiotics. Parameters of oropharyngeal and mucosal microflora, immune parameters of anti-infectious resistance (phagocytic activity, phagocytic index, neutrophilic index of digestion), as well as secretory immunoglobulin A levels in saliva were measured during the course of the illness. Results: In the patients of the main group under the combination therapy, there was a significantly more rapid elimination of respiratory symptoms and intoxication (p 0.05). Improvements of immune parameters of the anti-infectious resistance system were found only in the main group: the phagocytic index at 120 minutes after incubation of neutrophils was 4.26 ± 0.04 before treatment and 3.94 ± 0.09 after treatment (р < 0.05); the neutrophil digestion index, 5.70 ± 0.71 and10.83 ± 0.94 (р < 0.01), and secretory IgA level in saliva, 0.05 ± 0.03 and 0.125 ± 0.03 mcg/mL, respectively (р < 0.05). Conclusion: Inclusion of probiotic Florin forte into combination therapy of ARD patients promoted more rapid clinical recovery, improvement of biocenotic parameters in oropharyngeal and gut mucosa and an increase in anti-infectious resistance of the host.

Adipose-Derived Mesenchymal Stem Cells Restore Impaired Mucosal Immune Responses in Aged Mice.

It has been shown that adipose-derived mesenchymal stem cells (AMSCs) can differentiate into adipocytes, chondrocytes and osteoblasts. Several clinical trials have shown the ability of AMSCs to regenerate these differentiated cell types. Age-associated dysregulation of the gastrointestinal (GI) immune system has been well documented. Our previous studies showed that impaired mucosal immunity in the GI tract occurs earlier during agingthan is seen in the systemic compartment. In this study, we examined the potential of AMSCs to restore the GI mucosal immune system in aged mice. Aged (>18 mo old) mice were adoptively transferred with AMSCs. Two weeks later, mice were orally immunized with ovalbumin (OVA) plus cholera toxin (CT) three times at weekly intervals. Seven days after the final immunization, when fecal extract samples and plasma were subjected to OVA- and CT-B-specific ELISA, elevated levels of mucosal secretory IgA (SIgA) and plasma IgG antibody (Ab) responses were noted in aged mouse recipients. Similar results were also seen aged mice which received AMSCs at one year of age. When cytokine production was examined, OVA-stimulated Peyer’s patch CD4+ T cells produced increased levels of IL-4. Further, CD4+ T cells from the lamina propria revealed elevated levels of IL-4 and IFN-γ production. In contrast, aged mice without AMSC transfer showed essentially no OVA- or CT-B-specific mucosal SIgA or plasma IgG Ab or cytokine responses. Of importance, fecal extracts from AMSC transferred aged mice showed neutralization activity to CT intoxication. These results suggest that AMSCs can restore impaired mucosal immunity in the GI tract of aged mice.

Nasal fluid secretory immunoglobulin A levels in children with allergic rhinitis

ObjectivesThere is growing knowledge about the immunoregulatory and possibly preventative roles of immunoglobulin A (IgA) in allergic diseases. This study aimed to investigate secretory immunoglobulin A (SIgA) levels in the nasal fluid of children who were either being treated for their allergic rhinitis (AR) with intranasal mometasone furoate or were not receiving treatment. MethodsThe study population contained 55 children with persistent AR. Group I included 27 newly diagnosed AR patients not taking any medication and group II included 28 patients treated with intranasal steroids for at least 6 months. 27 healthy control subjects were also enrolled in the study. Total symptom scores (TSS) were calculated for each patient. Nasal secretions were obtained using a new modified polyurethane sponge absorption method, and samples were analysed by ELISA. ResultsThe median value for nasal fluid SIgA level in each group was 127.2μg/ml (interquartile range; 67.3–149.6) in group I, 133.9μg/ml (102.1–177.8) in group II and 299.8μg/ml (144.5–414.0) in the control group. Groups I and II both had statistically significant reductions in nasal fluid SIgA levels compared to the control group (p<0.001). However, there was no statistically significant difference between groups I and II (p=0.35). A statistically significant and negative correlation also existed between TSS and nasal fluid SIgA levels in both groups I and II (p=0.006, rho=−0.512 and p=0.01, rho=−0.481, respectively). ConclusionsSIgA levels in the nasal fluid are significantly reduced in children with AR independent of treatment and are negatively correlated with the TSS.

Detection of Allergen Specific Antibodies From Nasal Secretion of Allergic Rhinitis Patients.

PurposeAllergic rhinitis (AR) is a common and increasing disease in which Dermatophagoides (D.) farinae is one of the most common causative allergens. The aims of this study were to confirm the presence of locally produced antibodies to D. farinae in nasal secretions between nasal provocation test (NPT)-positive and -negative groups of AR patients, to evaluate their relationships with the levels of inflammatory mediators, and to determine adaptive and innate immune responses in nasal mucosa.MethodsSixty AR patients sensitive to house dust mites confirmed by skin prick test or serum specific IgE to D. farinae underwent NPT for D. farinae. Nasal packs were placed in both nasal cavities of the patients for 5 minutes to obtain nasal secretions after NPT. The levels of total IgE, specific IgE to D. farinae, eosinophil cationic protein (ECP), and tryptase in nasal secretions were detected by using ImmunoCAP. The levels of specific IgE, IgA, and secretory IgA antibodies to D. farinae in nasal secretions were measured by using ELISA. The levels of IL-8, VEGF, IL-25, and IL-33 were also measured by using ELISA.ResultsHigh levels of total IgE, specific IgE, specific IgA, and secretory IgA to D. farinae, as well as inflammatory mediators, such as ECP, IL-8, VEGF and tryptase, were detected in nasal secretions, although the differences were not statistically significant between the NPT-positive and NPT-negative groups. Levels of all immunoglobulins measured in this study significantly correlated with ECP, IL-8, and VEGF (P<0.05), but not with tryptase (P>0.05). IL-33 and IL-25 were also detected, and IL-25 level significantly correlated with IL-8 (r=0.625, P<0.001).ConclusionsThese findings confirmed the presence of locally produced specific antibodies, including D. farinae-specific IgE and IgA, in nasal secretions collected from D. farinae-sensitive AR patients in both the NPT-positive and NPT-negative groups, and close correlations were noted between antibodies and nasal inflammatory mediators, including such as ECP, IL-8 and VEGF, indicating that locally produced antibodies may be involved in the nasal inflammation of AR.

Protective efficacy and immune responses by homologous prime-booster immunizations of a novel inactivatedSalmonellaGallinarum vaccine candidate

PurposeSalmonella enterica serovar Gallinarum (SG) ghost vaccine candidate was recently constructed. In this study, we evaluated various prime-boost vaccination strategies using the candidate strain to optimize immunity and protection efficacy against fowl typhoid.Materials and MethodsThe chickens were divided into five groups designated as group A (non-immunized control), group B (orally primed and boosted), group C (primed orally and boosted intramuscularly), group D (primed and boosted intramuscularly), and group E (primed intramuscularly and boosted orally). The chickens were primed with the SG ghost at 7 days of age and were subsequently boosted at the fifth week of age. Post-immunization, the plasma IgG and intestinal secretory IgA (sIgA) levels, and the SG antigen-specific lymphocyte stimulation were monitored at weekly interval and the birds were subsequently challenged with a virulent SG strain at the third week post-second immunization.ResultsChickens in group D showed an optimized protection with significantly increased plasma IgG, sIgA, and lymphocyte stimulation response compared to all groups. The presence of CD4+ and CD8+ T cells and monocyte/macrophage (M/M) in the spleen, and splenic expression of cytokines such as interferon γ (IFN-γ) and interleukin 6 (IL-6) in the immunized chickens were investigated. The prime immunization induced significantly higher splenic M/M population and mRNA levels of IFN-γ whereas the booster showed increases of splenic CD4+ and CD8+ T-cell population and IL-6 cytokine in mRNA levels.ConclusionOur results indicate that the prime immunization with the SG ghost vaccine induced Th1 type immune response and the booster elicited both Th1- and Th2-related immune responses.

The piglet as a model for studying dietary components in infant diets: effects of galacto-oligosaccharides on intestinal functions.

Prebiotic oligosaccharides, including galacto-oligosaccharides (GOS), are used in infant formula to mimic human milk oligosaccharides, which are known to have an important role in the development of the intestinal microbiota and the immune system in neonates. The maturation of the intestines in piglets closely resembles that of human neonates and infants. Hence, a neonatal piglet model was used to study the multi-faceted effect of dietary GOS in early life. Naturally farrowed piglets were separated from the mother sow 24-48 h postpartum and received a milk replacer with or without the addition of GOS for 3 or 26 d, whereafter several indicators of intestinal colonisation and maturation were measured. Dietary GOS was readily fermented in the colon, leading to a decreased pH, an increase in butyric acid in caecum digesta and an increase in lactobacilli and bifidobacteria numbers at day 26. Histomorphological changes were observed in the intestines of piglets fed a GOS diet for 3 or 26 d. In turn, differences in the intestinal disaccharidase activity were observed between control and GOS-fed piglets. The mRNA expression of various tight junction proteins was up-regulated in the intestines of piglet fed a GOS diet and was not accompanied by an increase in protein expression. GOS also increased defensin porcine β-defensin-2 in the colon and secretory IgA levels in saliva. In conclusion, by applying a neonatal piglet model, it could be demonstrated that a GOS-supplemented milk replacer promotes the balance of the developing intestinal microbiota, improves the intestinal architecture and seems to stimulate the intestinal defence mechanism.