Ex Vivo Immunomodulatory Effects of Lactobacillus-, Lacticaseibacillus-, and Bifidobacterium-Containing Synbiotics on Human Peripheral Blood Mononuclear Cells and Monocyte-Derived Dendritic Cells in the Context of Grass Pollen Allergy.

Sensing of the intestinal microbiota by the host immune system is important to induce protective immune responses. Hence, modification of the gut microbiota might be able to prevent or treat allergies, mediated by proinflammatory Th2 immune responses. The aim was to investigate the ex vivo immunomodulatory effects of the synbiotics Pollagen® and Kallergen®, containing the probiotic bacterial strains Lactobacillus, Lacticaseibacillus and Bifidobacterium, in the context of grass pollen allergy. Peripheral blood mononuclear cells (PBMCs) from grass pollen-allergic patients and healthy controls were stimulated with grass pollen extract (GPE) and synbiotics and Gata3 expression and cytokine secretion analyzed. Monocyte-derived dendritic cells (MoDCs) cells were matured in the presence of GPE and synbiotics, co-cultured with autologous naïve T cells and maturation markers and cytokine secretion analyzed. GPE stimulation of PBMCs from grass pollen-allergic patients resulted in a significant higher production of the Th2 cytokines IL-4, IL-5, IL-9 and IL-13 compared to healthy controls. Gata3+CD4+ T cell induction was independent of the allergic status. The synbiotics promoted IL-10 and IFN-γ secretion and downregulated the GPE-induced Th2-like phenotype. Co-culturing naïve T cells with MoDCs, matured in the presence of GPE and synbiotics, shifted the GPE-induced Th2 cytokine release towards Th1-Th17-promoting conditions in allergic subjects. The investigated synbiotics are effective in downregulating the GPE-induced Th2 immune response in PBMCs from grass pollen-allergic patients as well as in autologous MoDC-T cell stimulation assays. In addition to increased IL-10 release, the data indicates a shift from a Th2- to a more Th1- and Th17-like phenotype.

House dust microbiota in relation to adult asthma and atopy in a US farming population.

BACKGROUND: Bacterial exposure from house dust has been associated with asthma and atopy in children but whether these relationships are present in adults remains unclear. OBJECTIVE: We sought to examine associations of house dust microbiota with adult asthma, atopy, and hay fever. METHODS: Vacuumed bedroom dust samples from the homes of 879 participants (average age, 62 years) in the Agricultural Lung Health Study, a case-control study of asthma nested within a farming cohort, were subjected to 16S rRNA amplicon sequencing to characterize bacterial communities. We defined current asthma and hay fever using questionnaires and current atopy by blood specific IgE level > 0.70 IU/mL to 1 or more of 10 common allergens. We used linear regression to examine whether overall within-sample bacterial diversity differed by outcome, microbiome regression-based kernel association test to evaluate whether between-sample bacterial community compositions differed by outcome, and analysis of composition of microbiomes to identify differentially abundant bacterial taxa. RESULTS: Overall diversity of bacterial communities in house dust was similar by asthma status but was lower (P < .05) with atopy or hay fever. Many individual bacterial taxa were differentially abundant (false-discovery rate, <0.05) by asthma, atopy, or hay fever. Several taxa from Cyanobacteria, Bacteroidetes, and Fusobacteria were more abundant with asthma, atopy, or hay fever. In contrast, several taxa from Firmicutes were more abundant in homes of individuals with adequately controlled asthma (vs inadequately controlled asthma), individuals without atopy, or individuals without hay fever. CONCLUSIONS: Microbial composition of house dust may influence allergic outcomes in adults.

Pollen Allergy Suppression Effect by the Oral Administration of Acetic Acid Bacteria (Gluconacetobacter hansenii).

BACKGROUND/AIM: Gluconacetobacter hansenii (G. hansenii) is an acetic acid bacterium of vinegar production. Its anti-allergic effect on mice upon oral administration was examined. MATERIALS AND METHODS: The amount of LPS was measured by the Limulus reaction. Mice were sensitized by peritoneal and intranasal administration of cedar pollen and alum followed by oral administration of 30 or 150 mg/kg of heated G. hansenii cells. Pollen was administered intranasally to evaluate nasal symptoms, and at 8 weeks, IgE and IL-10 levels in blood were measured by ELISA. RESULTS: The amount of LPS in dried bacterial cells was 10.4±3.3 mg/g. In the cedar pollinosis model of mice, a significant reduction was observed in nose scratching of both groups administered with the bacterial cells (30, 150 mg/kg). CONCLUSION: G. hansenii contains LPS, and its oral administration showed an anti-allergic effect by a significant mitigation of the symptoms in a pollen allergy mouse model.

Probiotics modulate gut microbiota and health status in Japanese cedar pollinosis patients during the pollen season.

BACKGROUND: Japanese cedar pollinosis (JCP) is a challenging public health problem in Japan. Altered gut microbiota is associated with several diseases, including allergic diseases. However, only a few studies have focused on JCP and the underlying mechanisms for probiotic effects remain unclear. In addition, this study is the first observation of the correlation between the gut microbiota and blood lipid in JCP. METHODS: Faecal samples from JCP subjects were collected before and after treatment with (n = 14) and without (n = 11) LGG-TMC0356-fermented milk for 10 weeks. Gut microbiota composition was characterized from faecal DNA using sequencing of 16S rRNA genes. RESULTS: 16S rRNA-based operational taxonomic unit clustering of the microbiota revealed that LGG-TMC0356-fermented milk significantly altered gut microbiota after 10 weeks of milk consumption, and eight dominant genera of microbes were detected. During the JCP season, the Bacteroidetes/Firmicutes ratio, when compared to baseline, was significantly decreased in subjects at end of the study. Bacteroidetes showed positive correlation with LDL- and HDL-cholesterol levels, whereas Firmicutes showed negative correlation with total cholesterol, LDL- and HDL- cholesterol. CONCLUSIONS: The altered gut microbiota through supplementation of fermented milk containing the study probiotics may be a prospective target for protection against JCP, with beneficial effects on blood lipid levels.

Alternaria and Cladosporium calendar of Western Thrace: Relationship with allergic rhinitis symptoms.

OBJECTIVES/HYPOTHESIS: Alternaria and Cladosporium are the most important outdoor moulds. The aim of this study was to present fungal spore monitoring data, investigate the relationship of fungal counts with climate conditions, and to explore the clinical significance of Alternaria and Cladosporium species monitoring in allergic rhinitis (AR). STUDY DESIGN: Analytic observational study. METHODS: A 7-day volumetric trap was used to collect circulating Alternaria and Cladosporium fungi. Sixty-nine AR patients were studied and recorded their symptoms by Total 5 Symptoms Score (T5SS). Sensitization prevalence to fungi species was detected by skin prick tests. Monitoring data were correlated to climate conditions and patients symptoms score. RESULTS: Alternaria and Cladosporium were detected throughout the calendar year in ranges of 0 to 217 spores/m(3) and 6.5 to 1,600 spores/m(3) per day, respectively. Highest daily concentrations of both fungi were detected during the summer (73.9 ± 34.4 spores/m(3) for Alternaria and 595.8 ± 288.0 spores/m(3) for Cladosporium) and lowest during the winter (2.4 ± 3.0 spores/m(3) for Alternaria and 24.3 ± 15.7 spores/m(3) for Cladosporium). Both were positively correlated to mean daily temperature and negatively to relative humidity (all P < 0.001). Clinically, a strong significant correlation between T5SS and airborne fungi levels, both for Alternaria (r = 0.822, P = 0.001) and Cladosporium (r = 0.787, P = 0.002) species was observed. CONCLUSIONS: We found Cladosporium to be the most frequently detected airborne mould, whereas Alternaria was the most prevalent with regard to sensitization rate. Patients' symptoms score was significantly correlated to spore concentrations. Both fungi were clearly affected by climate factors, such as temperature and relative humidity. These findings are important in AR management. LEVEL OF EVIDENCE: N/A.

Seasonal allergic rhinitis affects sinonasal microbiota.

BACKGROUND: Microbes and allergens can stimulate the nasal mucosa, potentially leading to the development of acute bacterial rhinosinusitis (ABRS). This study was designed to determine if allergen exposure alters the sinonasal microbiome. METHODS: We performed a parallel observational study of healthy adults with seasonal allergic rhinitis (SAR; grass or tree, n = 20) or nonallergic subjects (n = 19). Microbiota specimens were obtained by endoscopy from the middle meatus and vestibule before and during the relevant season and were analyzed by terminal restriction fragment length polymorphism analysis. Differences in bacterial microbiota were assessed by standard ecological measures of bacterial diversity. Quality of life and symptom scores were recorded, and nasal lavages for eosinophils were performed. RESULTS: SAR subjects had increased nasal symptoms in season, impaired disease-specific quality of life, and increased nasal eosinophils, compared with no changes in nonallergic subjects. During the season, SAR subjects had a significantly greater variety of organisms in the middle meatus compared with nonallergic subjects (p < 0.036) and increased bacterial diversity (Shannon index, p < 0.013). We found a significant positive correlation between bacterial diversity in the middle meatus during the season and the nasal lavage eosinophil count of SAR subjects. There were no significant changes in the nasal vestibule (p > 0.05, all comparisons). CONCLUSION: The interaction of allergy and microbiota may affect the sinonasal physiology, with broad implications for several airway diseases. Characterization of the specific organisms involved using next-generation sequencing may clarify the relationship between allergic inflammation and ABRS. This finding may help explain why allergic inflammation predisposes to ABRS.

C-type lectin receptors mRNA expression in patients with otitis media with effusion.

OBJECTIVE: The role of C-type lectin receptor, a type of pattern recognition receptor, in otitis media with effusion (OME) is unclear. We assayed the levels of expression of C-type lectin receptor mRNA in children with OME and evaluated its relationship to the presence of bacteria, accompanying diseases, and characteristics of exudates. SUBJECTS AND METHODS: The study population consisted of 73 children with OME who had undergone ventilating tube insertion. The levels of expression of Dectin-1, MR1, MR2, DC-SIGN, Syk, Card-9, Bcl-10, Malt-1, Src, Dec-205, Galectin-1, Tim-3, Trem-1, and DAP-12 mRNA in middle ear effusion were determined by real-time PCR. The level of expression of each mRNA was correlated with the presence of bacteria, accompanying diseases, and exudates characteristics. RESULTS: The levels of expression of C-type lectin receptor mRNAs were not associated with bacterial presence or exudates characteristics (p>0.05 each). Levels of expression, however, were significantly higher in patients with sinusitis, adenoid vegetation or adenoiditis, and allergic rhinitis (p<0.05 each). CONCLUSIONS: Levels of expression of C-type lectin receptor mRNA may be associated with the pathogenesis of OME, being significantly higher in patients with than without accompanying sinusitis, adenoid vegetation or adenoiditis, and allergic rhinitis.

[Analysis of bacterial culture and cytology in nasal smear of Japanese ceder pollinosis in pre-season and in season].

BACKGROUND: Previously, we reported that nasal symptom appeared in some patients of Japanese cedar pollinosis (JCP) before season. Over than 50% of them had neutrophils (no eosinophil) in their nasal smear (less than 10%; eosinophils), and Staphylococcus aureus or epidermis by culture were detected in more than 90% of them. We thought the attack caused from rhinitis by Staphylococcus. METHOD: For more details of these bacterium in JCP, we enforced bacterial culture in the nose and cytology in the nasal smear in asymptomatic (PreAs), symptomatic in pre-season (PreSy), and symptomatic in season (InSy) of JCP in 2011. RESULTS: Positive rates of Staphylococcus aureus were 79%, 75%, 53%, Staphylococcus epidermis; 15%, 10%, 16%, Moraxella catarrhalis; 9%, 25%, 19%, and Streptococcus Pneumonia; 7%, 12%, 16% in PreAs (n=53), PreSy (60), and InSy (70) respectively. Cytology showed that 45% of PreAs (n=53), 65% of PreSy (60) with statistical higher in latter group, and 8% of InSy (72) were only neutrophils in nasal smear. Six% of PreAs, 2% of PreSy, and 51% of InSy were eosinophils. Forty-three% of PreAs with statistical higher than 22% of PreSy and 7% of InSy were nasal cell free. CONCLUSION: These results mean the inflammation by Staphylococcus species or Moraxella catarrhalis might involve symptom appearance in pre-season.

Assessment of biofilm by nasal cytology in different forms of rhinitis and its functional correlations.

BACKGROUND: Recently, it has been reported that nasal cytology in light microscopy can identify biofilms, which appear as cyan-stained "Infectious Spots". We assessed by the same method and in the same population, the presence of biofilms in different nasal disorders, and estimated if a correlation with the functional grade of obstruction existed. METHODS: Subjects suffering from different nasal disorders, after a detailed clinical history and ENT examination, underwent nasal fibroendoscopy, skin prick test, rhinomanometry and nasal cytology. The presence of biofilm was linked to the type ofdisease and to the grade of obstruction. RESULTS: Among 1,410 subjects previously studied, the infectious spot was found in 107 patients (7.6%), and this percentage reached 55.4% in subjects with cytologic signs of infectious rhinitis (presence of bacteria/fungi). Biofilms were largely more frequent in patients with adenoid hypertrophy (57.4%), followed by nasal polyposis (24%), chronic rhinosinusitis (9.5%) and non-allergic rhinitis (7.6%). Nasal cytology was normal in the remaining patients, where no infectious spot was detectable. Statistical analysis showed that nasal resistances were significantly higher in presence of biofilms in patients with adenoid hypertrophy (p = 0.003), nasal polyposis (p < 0.001), chronic rhinosinusitis (p = 0.018) and septal deviation (p = 0.001). CONCLUSION: The results demonstrate that biofilm is present not only in infectious rhinitis, but also in inflammatory and/or immune-mediated diseases. The presence of biofilms significantly correlates with the degree of nasal obstruction as assessed by rhinomanometry.

Update on immune mechanisms associated with sublingual immunotherapy: practical implications for the clinician.

Sublingual immunotherapy (SLIT) is established as a safe and efficacious treatment for patients with type I respiratory allergies. The ability of SLIT to elicit antigen (allergen)-specific tolerance is linked to the peculiar biology of oral antigen-presenting cells. In the absence of danger signals, Langerhans cells, myeloid dendritic cells, and macrophages located in oral tissues, tonsils, and draining cervical lymph nodes are biased toward the induction of T(H)1 and IL-10-producing CD4(+) regulatory T cells, thus supporting tolerance as opposed to inflammation. Sublingual administration does not lead to any detectable systemic exposure of intact allergens nor to IgE neosensitization. Oral tissues contain limited numbers of mast cells located in submucosal areas, thereby explaining the well-established safety profile of SLIT, with mostly local but rare systemic reactions. The induction of CD4(+) regulatory T cells and blocking anti-inflammatory IgGs or IgAs are considered important for tolerance induction after SLIT. Specific molecular signatures associated with tolerogenic dendritic cells were recently reported during the onset of SLIT efficacy in the peripheral blood of patients exhibiting clinical benefit. Collectively, these observations confirm the induction of strong allergen-specific suppressive/tolerogenic immune responses during SLIT and pave the ground for the identification of biomarkers of efficacy. Practical implications of this emerging scientific knowledge are presented (1) to support the rational design of second-generation sublingual vaccines based on purified allergens, vector systems and/or adjuvants and (2) to help the clinician in decision making during his/her practice.

Streptococcus pyogenes upper respiratory infection and atopic conditions other than asthma: a retrospective cohort study.

BACKGROUND: Patients with asthma have an increased risk of Streptococcus pyogenes infection compared with those without asthma. It is unknown whether this is true for children with other atopic conditions such as atopic dermatitis or allergic rhinitis. AIMS: To determine the risk of developing S. pyogenes infections of the upper respiratory tract in children and adolescents with atopic dermatitis and/or allergic rhinitis. METHODS: We conducted a retrospective cohort study that followed a convenience sample of 340 healthy children. Atopic dermatitis or eczema and allergic rhinitis or hay fever were determined based on a physician diagnosis documented in medical records. All laboratory test results of cultures, rapid antigen detection, and polymerase chain reaction tests for S. pyogenes infections during the first 18 years of life were collected to compare the incidence of S. pyogenes infections between children with and without a physician diagnosis of atopic conditions. A Poisson regression was fit to determine the association between asthma and S. pyogenes infections, controlling for other covariates including asthma. RESULTS: Of the 340 subjects, 327 were eligible for the study. Of these 327 subjects, 143 (44%) had atopic conditions other than asthma. The incidence of S. pyogenes infections in children with atopic conditions other than asthma and those without atopic conditions was 0.24 per person-year and 0.18 per person-year, respectively. The adjusted risk ratios for allergic rhinitis and atopic dermatitis were 1.36 (95% CI 1.07 to 1.66, p=0.011) and 1.30 (95% CI 0.98 to 1.71, p=0.06), respectively, controlling for asthma and other covariates. CONCLUSIONS: In addition to asthma, allergic rhinitis but not atopic dermatitis is associated with an increased risk of S. pyogenes upper respiratory tract infections.

Microbiology of sinusitis: does allergy or endoscopic sinus surgery affect the microbiologic flora?

PURPOSE OF REVIEW: To explore the role of bacteria in the pathogenesis of chronic rhinosinusitis and how the presence of allergies or endoscopic sinus surgery may affect the bacterial flora. RECENT FINDINGS: As our understanding of the etiologies of chronic rhinosinusitis continues to evolve, the role of bacteria as either a primary or an exacerbating factor remains controversial. It is clear that healthy paranasal sinuses are sterile, but in chronic rhinosinusitis bacteria are often present. It has also been shown that the bacteria found in chronic rhinosinusitis vary significantly from those found in acute rhinosinusitis. In patients with allergic rhinitis an association has been found with higher carriage rates of Staphylococcus aureus. The significance of this is unclear but may be related to bacterial superallergen production leading to TH2-mediated inflammation. Endoscopic sinus surgery does not appear to change the bacterial flora, though it may decrease the presence of bacterial biofilms in sinus cavities. SUMMARY: Bacteria are likely to play a role in both the development and the exacerbation of chronic rhinosinusitis. Further studies are needed to determine in which subtypes of chronic rhinosinusitis bacteria play the greatest roles and how our treatments can be individualized to improve patient outcomes.

Alternaria sensitization and allergic rhinitis with or without asthma in the French Six Cities study.

BACKGROUND: Allergic sensitization to Alternaria has been related to asthma in various studies, but its association with allergic rhinitis is still controversial. OBJECTIVES: The aim of this study was to assess at the population level the relationships in childhood between Alternaria sensitization and 'past-year rhinoconjunctivitis' (PYRC), 'ever hay fever' (EHF) and 'ever allergic rhinitis caused by allergens other than pollens' (EAR) according to the presence or the absence of asthma. METHODS: This study is part of the Six Cities Study, the French contribution to the International Study of Asthma and Allergies in Childhood (ISAAC) Phase II. Children underwent skin prick test (SPT) to Alternaria and parents filled a standardized medical questionnaire. RESULTS: Some 6726 children with a mean age of 10 years were examined. The overall prevalence of Alternaria sensitization was 2.8%, 0.8% for monosensitization. Prevalences of symptoms in sensitized children were 27.7% for PYRC, 27.0% for EHF and 30.4% for EAR. Adjusted Odds Ratios (OR) between Alternaria sensitization and allergic rhinitis phenotypes were 2.34 (95% confidence interval: 1.51-3.63) for PYRC, 2.40 (1.65-3.50) for EHF and 2.95 (2.05-4.23) for EAR. The relationship still remained in the case of monosensitization to Alternaria for both PYRC and EAR when excluding the asthmatic children [OR = 3.87 (1.54-9.78) and 2.88 (1.10-7.55) respectively]. CONCLUSION: In our population-based sample of children, we found a link between Alternaria sensitization and allergic rhinitis, independently of asthma, which is compatible with the mechanisms of deposition of Alternaria in the upper airways.

Effects of daily intake of Lactobacillus paracasei strain KW3110 on Japanese cedar pollinosis.

Japanese cedar pollinosis is an important contributor to allergic rhinitis in Japan. Lactobacillus may be useful as an immunomodulator and is used widely as a foodstuff. The purpose of the study was to examine the effects of daily intake of the Lactobacillus paracasei strain KW3110 in patients with cedar pollinosis. The effects of daily intake of KW3110 in patients with cedar pollinosis were investigated in 126 patients who received KW3110 or a placebo in a double-blind study. The study began 1 month before the start of the pollen season and lasted for 3 months. A significant reduction of nasal symptoms and the serum level of eosinophil cationic protein and improvement of quality of life scores occurred in the patients who received KW3110 when pollen scattering was low. However, the effects were limited during the peak period of pollen scattering. Intake of KW3110 may reduce allergic inflammation, but the effect is limited.

Specific probiotics alleviate allergic rhinitis during the birch pollen season.

AIM: To investigate whether birch pollen allergy symptoms are linked with gut microbiota changes and whether probiotics have an effect on these. METHODS: Forty seven children with confirmed birch pollen allergy were randomized to receive either a probiotic combination of Lactobacillus acidophilus (L. acidophilus) NCFM (ATCC 700396) and Bifidobacterium lactis (B. lactis) Bl-04 (ATCC SD5219) or placebo in a double-blind manner for 4 mo, starting prior to onset of the birch pollen season. Symptoms were recorded in a diary. Blood samples were taken for analysis of cytokines and eosinophils. Fecal samples were analysed for microbiota components, calprotectin and IgA. Nasal swabs were taken for analysis of eosinophils. RESULTS: The pollen season induced a reduction in Bifidobacterium, Clostridium and Bacteroides which could not be prevented by the probiotic intervention. During the intervention, significantly higher numbers of B. lactis 11.2 x 10(7) +/- 4.2 x 10(7) vs 0.1 x 10(7) +/- 0.1 x 10(7) bacteria/g feces (P < 0.0001) and L. acidophilus NCFM 3.5 x 10(6) +/- 1.3 x 10(6) vs 0.2 x 10(6) +/- 0.1 x 10(6) bacteria/g feces (P < 0.0001) were observed in the probiotic group compared to the placebo group. During May, there was a tendency for fewer subjects, (76.2% vs 95.2%, P = 0.078) to report runny nose, while during June, fewer subjects, 11.1% vs 33.3%, reported nasal blocking in the probiotics group (P = 0.101). Concomitantly, fewer subjects in the probiotic group had infiltration of eosinophils in the nasal mucosa compared to the placebo group, 57.1% vs 95% (P = 0.013). Eye symptoms tended to be slightly more frequent in the probiotic group, 12.5 d [interquartile range (IQR) 6-18] vs 7.5 d (IQR 0-11.5) (P = 0.066) during May. Fecal IgA was increased in the placebo group during the pollen season; this increase was prevented by the probiotics (P = 0.028). CONCLUSION: Birch pollen allergy was shown to be associated with changes in fecal microbiota composition. The specific combination of probiotics used was shown to prevent the pollen-induced infiltration of eosinophils into the nasal mucosa, and indicated a trend for reduced nasal symptoms.

Distribution of different species of the Bacteroides fragilis group in individuals with Japanese cedar pollinosis.

We investigated associations of species of the Bacteroides fragilis group with Japanese cedar pollinosis (JCPsis). Cell numbers of Bacteroides fragilis and Bacteroides intestinalis were significantly higher in JCPsis subjects than in non-JCPsis subjects before the pollen season. They correlated positively with both symptom scores and JCPsis-specific immunoglobulin E levels.

Fungus culture of the nasal secretion of chronic rhinosinusitis patients: seasonal variations in Daegu, Korea.

BACKGROUND: The role of fungi in the pathogenesis of chronic rhinosinusitis (CRS) is being increasingly recognized. The presences of fungal spores are a necessary condition for fungal sinusitis, and fungal spore counts vary widely, both geographically and seasonally. The aim of this study was to characterize fungal seasonal variation in the nasal cavity of CRS patients and to compare it with histologic types of nasal polyps. METHODS: One hundred eight patients with a clinical diagnosis of CRS with polyps were enrolled in this study for 1 year. Nasal secretions were obtained by aspiration and cultured to identify fungi. Nasal polyps were divided into four histologic types: eosinophilic, fibroinflammatory, seromucinous, and atypical. RESULTS: Positive cultures for fungi were obtained in 60 of 108 (63.0%) CRS patients, and 28 of 45 (62.2%) normal volunteers. Fungus culture rates were higher during summer and fall. Cladosporium, Aspergillus, Alternaria, and Penicilium were frequently isolated from CRS patients and normal volunteers. The most common histologic type was the fibroinflammatory type (60.2%), although in the summer and fall the proportion of the eosinophilic type increased. CONCLUSION: Fungi were commonly cultured during a hot and humid environment, during summer and fall, and cultured organism types were found to be seasonal dependent. Histologic types of nasal polyps appear to be associated with fungal culture rate, organism type, and seasonal variations.

Influence of Bifidobacterium longum BB536 intake on faecal microbiota in individuals with Japanese cedar pollinosis during the pollen season.

It has been reported that intake of yogurt or powder supplemented with the Bifidobacterium longum BB536 probiotic strain alleviated subjective symptoms and affected blood markers of allergy in individuals with Japanese cedar pollinosis (JCPsis) during the pollen seasons of 2004 and 2005, based on randomized, double-blind, placebo-controlled trials. Furthermore, the 2004 study found that intestinal bacteria such as the Bacteroides fragilis group significantly fluctuated during the pollen season in JCPsis individuals and intake of BB536 yogurt tended to suppress these fluctuations. The present study investigated faecal microbiota to examine whether any changes occurred during the pollen season and whether any influence was exerted by intake of BB536 powder in the 2005 pollen season, which happened to be a heavy season, to confirm the 2004 findings and to evaluate the relationship of microbiota with symptom development. In a randomized, double-blind, placebo-controlled trial, 44 JCPsis subjects received BB536 or a placebo for 13 weeks during the pollen season. Another 14 Japanese cedar pollen (JCP)-specific IgE negative healthy subjects received placebo for the same period. Faecal samples were collected before (week 0), during (weeks 4, 8 and 13) and after (week 17) intervention, and out of JCP season (week 28). Faecal microbiota were analysed using terminal-RFLP (T-RFLP) and real-time PCR methods. Principal component analysis based on T-RFLP indicated distinct patterns of microbiota between healthy subjects and JCPsis subjects in the placebo group, but an intermediate pattern in the BB536 group at week 13, the last stage of the pollen season. The coordinate of principal component 1 at week 13 correlated with composite scores of JCPsis symptoms recorded during the pollen season. Faecalibacterium prausnitzii and the Bacteroides fragilis group were identified as the main contributors to microbiotal fluctuations. Real-time PCR indicated that BB536 intake suppressed increases in the Bacteroides fragilis group compared with the placebo group (P <0.05). These results suggest that faecal microbiota in JCPsis subjects, but not healthy subjects, fluctuate at the end of the pollen season and that BB536 intake plays a role in maintaining normal microbiota.

Fluctuation of fecal microbiota in individuals with Japanese cedar pollinosis during the pollen season and influence of probiotic intake.

BACKGROUND: We have previously reported the results of a randomized, double-blind, placebo-controlled trial that found the intake of yogurt supplemented with a probiotic strain, Bifidobacterium longum BB536, alleviates symptoms and affects blood parameters in individuals with Japanese cedar pollinosis (JCPsis) during the pollen season. OBJECTIVE: In the present study, fecal microbiota were investigated to examine whether any changes occur during the pollen season and whether any influence is exerted by probiotic intake. METHODS: Yogurt either with BB536 (BB536 yogurt) or without BB536 (placebo yogurt) was administered for 14 weeks at 2 x 100 g per day to 40 subjects (17 men, 23 women) with a clinical history of JCPsis. Fecal samples were obtained from 23 subjects (placebo group, n=13; BB536 group, n=10) before and during the intervention (weeks 4, 9 and 13) and fecal microbiota were analyzed using terminal-restriction fragment length polymorphism and real-time polymerase chain reaction (PCR) methods. RESULTS: From the fluctuation patterns of terminal-restriction fragments, the Bacteroides fragilis group and bifidobacteria were among the species that changed most with pollen dispersion. Real-time PCR analyses indicated that the cell numbers of the B fragilis group increased significantly along with pollen dispersion in both BB536 and placebo groups. Cell numbers of bifidobacteria were significantly higher in the BB536 group compared with the placebo group (P < .05 at weeks 4 and 9). The ratio of cell numbers of the B fragilis group to bifidobacteria increased significantly during the pollen season in the placebo group (P < .01 at weeks 9 and 14), but not in the BB536 group. An in vitro study using peripheral blood mononuclear cells from JCPsis subjects indicated that strains of the B fragilis group induced significantly more helper T cell (T(H)) type2 cytokines (interleukin [IL]-6) but fewer T(H)1 cytokines (IL-12 and interferon) compared with those of bifidobacteria. CONCLUSIONS: These results suggest a relationship between fluctuation in intestinal microbiota and pollinosis allergy. Furthermore, intake of BB536 yogurt appears to exert positive ihfluences on the formation of anti-allergic microbiota.