Immune Function of Probiotic Bacterial DNA and Their Advanced Application to Food Immunology

The toll-like receptor (TLR) family plays an important role in host defense through recognizing bacterial pathogen-associated molecular patterns. The induction of the host defense system begins with the recognition of pathogens. To date, 13 types of TLR, which function as a kind of pathogen sensor, have been identified. Our recent research on the physiological function of food products has investigated the immunoregulatory effects of probiotic lactic acid bacteria (LAB) via TLR. On the basis of our study, efforts have been made to develop a molecular immunoassay system for probiotic LAB and find novel immunostimulatory DNA sequences from probiotics and high potential immunobiotic LAB strains via TLR9 signaling. These findings may provide important clues at the molecular level on TLR signal transduction pathways and recognition mechanisms for the ligands. In addition to identifying immunoregulatory factor immunogenics from LAB, a better understanding of intestinal immune regulation through cytokine networks holds out promise for basic food immunology research and the development of immunobiotic foods/feeds to prevent specific diseases.OtherShinshu University International Symposium 2010 : Sustainable Agriculture and Environment : Asian Networks II 　信州大学国際シンポジウム2010 : 持続的農業と環境 : アジアネットワークII ― アジアネットワークの発展をめざして―. 信州大学農学部, 2010, 93-97conference pape

Applications of Genetically Modified Immunobiotics with High Immunoregulatory Capacity for Treatment of Inflammatory Bowel Diseases

Inflammatory bowel diseases (IBDs), including ulcerative colitis and Crohn's disease, are chronic inflammatory diseases characterized by dysregulated immune responses of the gastrointestinal tract. In recent years, the incidence of IBDs has increased in developed nations, but their prophylaxis/treatment is not yet established. Site-directed delivery of molecules showing anti-inflammatory properties using genetically modified (gm) probiotics shows promise as a new strategy for the prevention and treatment of IBD. Advantages of gm-probiotics include 1) the ability to use bacteria as a delivery vehicle, enabling safe and long-term use by humans, 2) decreased risks of side effects, and 3) reduced costs. The intestinal delivery of anti-inflammatory proteins such as cytokines and enzymes using Lactococcus lactis has been shown to regulate host intestinal homeostasis depending on the delivered protein-specific machinery. Additionally, clinical experience using interleukin 10-secreting Lactococcus lactis has been shown to be safe and to facilitate biological containment in IBD therapy. On the other hand, some pre-clinical studies have demonstrated that gm-strains of immunobiotics (probiotics strains able to beneficially regulate the mucosal immunity) provide beneficial effects on intestinal inflammation as a result of the synergy between the immunoregulatory effects of the bacterium itself and the anti-inflammatory effects of the delivered recombinant proteins. In this review, we discuss the rapid progression in the development of strategies for the prophylaxis and treatment of IBD using gm-probiotics that exhibit immune regulation effects (gm-immunobiotics). In particular, we discuss the type of strains used as delivery agents.ArticleFrontiers in Immunology. 8:22 (2017)journal articl

Development of a New Treatment Modality for Lung Diseases That Uses Innovative Fine Droplet Drying (fdd) Technology Offering Inhalable Nano/microparticles-incorporated Therapeutic Agents.

ArticleImpact. 2019(3): 56-58. (2019)journal articl

Anti-allergic effects of a mixture of Saccharomyces cerevisiae and its specific goat's milk immunoglobulin G rich fraction on ovalbumin sensitized BALB/c mice

In this study, we investigated the effects of oral ingestion of Saccharomyces (S.) cerevisiae and its specific immunoglobulin (Ig) G-rich fraction, prepared from goat's milk, on immune function in an allergic rhinitis mouse model, ovalbumin (OVA)-sensitized BALB/c mice. Sneezing activity in mice that were administered a feed containing a mixture of S. cerevisiae and its specific goat's milk IgG-rich fraction (mixture group) was significantly lower than that in mice administered the mixture-free feed (control group) at age 11 and 12 weeks, although the activity in mice that were given a feed containing S. cerevisiae (yeast group) was similar to that of the control group. We found that the ratio of spleen interferon-gamma(+)CD4(+) cell/interleukin (IL)-4(+)CD4(+) cell was higher in the mixture and yeast groups than the control group. The number of spleen CD80(+)CD11b(+) cells was significantly lower in the mixture and yeast groups than the control group. In contrast, the numbers of spleen CD122(+)CD8 alpha(+) and IL-10(+)CD4(+) cells were significantly higher in the mixture group than the control and yeast groups while the numbers of IgE(+)B220(+) and Fc epsilon RI alpha(+)CD117(+) cells were significantly lower in the mixture group. These results suggested that the mixture of S. cerevisiae and its specific goat's milk IgG-rich fraction reduced OVA-induced allergic symptoms in BALB/c mice via the induction of suppressor T cells and the reduction of Ig E-producing cells and high affinity IgE receptor-positive mast cells rather than the adjustment of the Th1/Th2 balance.ArticleMILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL. 66(1): 7-10(2011)journal articl

Effect of Probiotics/Prebiotics on Cattle Health and Productivity

Probiotics/prebiotics have the ability to modulate the balance and activities of the gastrointestinal (GI) microbiota, and are, thus, considered beneficial to the host animal and have been used as functional foods. Numerous factors, such as dietary and management constraints, have been shown to markedly affect the structure and activities of gut microbial communities in livestock animals. Previous studies reported the potential of probiotics and prebiotics in animal nutrition; however, their efficacies often vary and are inconsistent, possibly, in part, because the dynamics of the GI community have not been taken into consideration. Under stressed conditions, direct-fed microbials may be used to reduce the risk or severity of scours caused by disruption of the normal intestinal environment. The observable benefits of prebiotics may also be minimal in generally healthy calves, in which the microbial community is relatively stable. However, probiotic yeast strains have been administered with the aim of improving rumen fermentation efficiency by modulating microbial fermentation pathways. This review mainly focused on the benefits of probiotics/prebiotics on the GI microbial ecosystem in ruminants, which is deeply involved in nutrition and health for the animal.ArticleMICROBES AND ENVIRONMENTS. 30(2):126-132 (2015)journal articl

Oral administration of a fruiting body extract of Boletopsis leucomelas enhances intestinal IgA production in LPS-challenged mice

The present study showed that a hot water extract of the fruiting body of the edible mushroom Boletopsis leucomelas, known as 'Kurokawa' Japanese, strongly stimulated IgA-production in mouse spleen cells in our screening experiment. The in vivo study was also conducted with the objective of enhancing adaptive immune response by oral administration of the hot water extract of B. leucomelas (BLE) in lipopolysaccharide (LPS)-challenged mice. The mice were fed a standard diet with or without 0.16% BLE. The mice were also orally administered sodium phosphate buffer or LPS weekly at days 7, 14 and 21. Results indicated that LPS-specific serum IgG, IgM and IgA were increased in the BLE diet group compared to the standard diet group. Interestingly, intestinal total IgA and LPS-specific IgA were significantly increased in the BLE diet group. Moreover, the <1000 Da, 5000-10,000 Da phenol sulphate-positive molecules from BLE showed significant IgA-producing activity.ArticleFOOD AND AGRICULTURAL IMMUNOLOGY. 25(4):510-522 (2014)journal articl

Myogenetic oligodeoxynucleotide (myoDN) recovers the differentiation of skeletal muscle myoblasts deteriorated by diabetes mellitus.

Skeletal muscle wasting in patients with diabetes mellitus (DM) is a complication of decreased muscle mass and strength, and is a serious risk factor that may result in mortality. Deteriorated differentiation of muscle precursor cells, called myoblasts, in DM patients is considered to be one of the causes of muscle wasting.We recently developed myogenetic oligodeoxynucleotides (myoDNs), which are 18-base single-strand DNAs that promote myoblast differentiation by targeting nucleolin. Herein, we report the applicability of a myoDN, iSN04, to myoblasts isolated from patients with type 1 and type 2 DM. Myogenesis of DM myoblasts was exacerbated concordantly with a delayed shift of myogenic transcription and induction of interleukins. Analogous phenotypes were reproduced in healthy myoblasts cultured with excessive glucose or palmitic acid, mimicking hyperglycemia or hyperlipidemia. iSN04 treatment recovered the deteriorated differentiation of plural DM myoblasts by downregulating myostatin and interleukin-8 (IL-8). iSN04 also ameliorated the impaired myogenic differentiation induced by glucose or palmitic acid. These results demonstrate that myoDNs can directly facilitate myoblast differentiation in DM patients, making them novel candidates for nucleic acid drugs to treat muscle wasting in patients with DM.ArticleFrontiers in Physiology. 12 : 679152 (2021)journal articl

Swine Intestinal Immunity via Toll-like Receptors and Its Advanced Application to Food Immunology

Recent interest has focused on the importance of intestinal immunity for the host defense, but to date, not much has been known about the underlying mechanisms. Toll-like receptor (TLR) family plays an important role in the defense through recognizing bacterial pathogen associated molecular patterns (PAMPs). Our research on the bioregulatory function of food products has investigated the immunoregulatory effects of lactic acid bacteria (LAB) via TLRs. Studies in swine, which is expected as a human model, have been examined intestinal immunoregulation by the LAB. Our research has now demonstrated modulation of intestinal immunity mediated by TLRs in Peyer\u27s patches and the mesenteric lymph nodes. On the basis of our study, efforts have also been made to develop an immunoassay system for immunobiotic LAB DNA and cell wall components to evaluate immunoregulation by the LAB via TLRs. The findings in our research activities may provide important clues at the molecular level on TLR signal transduction pathways and recognition mechanisms. They also provide impetus to further delineate the activation mechanism of the innate immune response. In addition, identification of biofactors from LAB with immunoactivity, and better understanding of cytokine induction and intestinal immune regulation hold promise in basic research and development of "immunobiotic foods" to prevent specific diseases

Intratracheally Therapeutic Option for COPD: A Potential Usage of the Therapeutic Microbe for Delivering Specific Protein to the Lungs

Currently, inhaled therapy using corticosteroids and/or bronchodilators is the major established treatment for chronic obstructive pulmonary disease (COPD). The topic to be covered in this chapter is the recently developed experimental approach using biologically active molecules secreted by the live genetically modified lactic acid bacteria (gmLAB). The strategy to use gmLAB as a therapeutic/delivering tool targeting disease-specific active molecules/cites is proceeding. The role of inflammation and oxidative stress in COPD development is a valid target point. Heme oxygenase (HO)-1 as an anti-inflammatory and antioxidative stress molecule has been examined to attenuate the lung function decline and inflammation in the murine model of COPD. Recently, HO-1-secreting gmLAB as a tool for targeting inflammatory diseases has been developed and examined in several disease models including COPD. When administered intratracheally, the gmLAB showed migration to the peripheral lung and overexpression of anti-inflammatory/oxidative HO-1 in both lung and serum, protecting the lung from COPD development