Masahiko Takeshita

Efficacy of oral administration of heat-killed probiotics from Mongolian dairy products against influenza infection in mice: Alleviation of influenza infection by its immunomodulatory activity through intestinal immunity

Some probiotics possess immunomodulatory activities and have been used as complementary and alternative medicines. We previously found that 10 lactic acid bacteria (LAB) strains isolated from traditional Mongolian dairy products showed probiotic potential in vitro. In this study, we assessed the immunomodulatory activity of 10 LABs on influenza virus (IFV) infection in relation to their efficacies in IFV-infected mice. In an intranasal IFV infection model in mice, oral administration of boiled Lactobacillus plantarum 06CC2 strain (20mg/mouse), one of the 10 LABs, twice daily for 10 days starting two days before infection was significantly effective in protecting the body weight loss of infected mice, reducing virus yields in the lungs on days 2, 4, and 6 after infection, and prolonging survival times without toxicity. The total numbers of infiltrated cells in the bronchoalveolar lavage fluid (BALF), especially macrophages and neutrophils, were significantly reduced by 06CC2 administration on day 2. On day 2, tumor necrosis factor (TNF)-α production in BALF was also reduced significantly, but interferon-α, interleukin-12, and interferon-γ productions were augmented and natural killer (NK) cell activity was significantly elevated. Furthermore, the gene expressions of interleukin-12 receptor and interferon-γ in Peyers patches were augmented by 06CC2 administration on day 2. Thus, 06CC2 was suggested to alleviate influenza symptoms in mice in correlation with the augmentation of NK cell activity associated with the enhancement of interferon-α and Th1 cytokine productions through intestinal immunity and the reduction of TNF-α in the early stage of infection.

The investigation of probiotic potential of lactic acid bacteria isolated from traditional Mongolian dairy products

The aims of this study were to investigate the diversity of lactic acid bacteria (LAB) isolated from traditional Mongolian dairy products, and to estimate the probiotic potential of the isolated strains. We collected 66 samples of the traditional Mongolian dairy products tarag (n = 45), airag (n = 7), aaruul (n = 8), byasulag (n = 1) and eezgii (n = 5), from which 543 LAB strains were isolated and identified based on 16S ribosomal DNA sequence. The predominant species of those products were Lactobacillus (L.) delbrueckii ssp. bulgaricus, L. helveticus, L. fermentum, L. delbrueckii ssp. lactis and Lactococcus lactis ssp. lactis. However, we could not detect any LAB strains from eezgii. All LAB isolates were screened for tolerance to low pH and to bile acid, gas production from glucose, and adherence to Caco-2 cells. In vitro, we found 10 strains possess probiotic properties, and almost identified them as L. plantarum or L. paracasei subspecies, based on 16S ribosomal DNA and carbohydrate fermentation pattern. These strains were differentiated from each other individually by randomly amplified polymorphic DNA analysis. Additionally, it was notable that 6/10 strains were isolated from camel milk tarag from the Dornogovi province.

Effects of Oral Administration of Probiotics from Mongolian Dairy Products on the Th1 Immune Response in Mice

We investigated 10 lactic acid bacteria strains with probiotic potential prepared from Mongolian dairy products for their ability to induce T helper type-1 (Th1) cytokine production in mouse immune cells in vitro and in vivo. Among these strains, the Lactobacillus plantarum 06CC2 strain was effective in elevating the level of interleukin (IL)-12p40 in co-culture with J774.1 cells and the levels of IL-12 and interferon (IFN)-γ in co-culture with mouse spleen cells in vitro. Oral administration of this strain augmented the gene expression of IFN-γ and IL-12p40 and enlarged the population of CD4+, CD25+, and CD49b+ cells in the spleens of normal mice. It also significantly elevated the gene expression of IL-12 receptor β2 as well as IL-12p40 and IFN-γ in Peyers patches. Thus oral administration of strain 06CC2 was effective in inducing Th1 cytokine production activating the Th1 immune response associated with intestinal immunity in normal mice.

Augmentation of T helper type 1 immune response through intestinal immunity in murine cutaneous herpes simplex virus type 1 infection by probiotic Lactobacillus plantarum strain 06CC2

We previously found that Lactobacillus plantarum strain 06CC2 showed probiotic potential, and its oral administration effectively induced Th1 cytokine production and activated the Th1 immune response associated with intestinal immunity in mice. In this study, to evaluate its potential as a versatile oral adjuvant for treatment of viral infection, we assessed the immunomodulatory activity of 06CC2 on murine cutaneous herpes simplex virus type 1 (HSV-1) infection, in which a major immune defense system is a delayed-type hypersensitivity (DTH) reaction based on activation of the Th1 immune response, in relation to its oral efficacy for alleviation of herpetic symptoms. In the HSV-1 infection model, oral administration of 06CC2 (20mg/mouse) twice daily for seven days starting two days before infection was significantly effective in delaying the development of skin lesions in the early phase of infection and reducing virus yields in the brain on day 4 after infection. In addition, 06CC2 significantly augmented the DTH reaction to inactivated HSV-1 antigen and elevated interferon (IFN)-γ production by HSV-1 antigen from splenocytes. On day 2, natural killer (NK) cell activity was significantly elevated, and the elevation was still observed on day 4. Furthermore, gene expressions of interleukin-12 receptor β2 and IFN-γ in Peyers patches were augmented on day 4 by 06CC2 administration. Thus, 06CC2 was suggested to alleviate herpetic symptoms in mice in correlation with augmentation of the Th1 immune responses associated with NK cell activity through intestinal immunity. Strain 06CC2 may be a versatile oral adjuvant to activate Th1 immune response.

Lactobacillus paracasei strain 06TCa19 suppresses inflammatory chemokine induced by Helicobacter pylori in human gastric epithelial cells

Helicobacter (H.) pylori infection is an important risk factor for gastric cancer that causes gastric inflammation. Inflammatory chemokines such as interleukin (IL)-8 and regulated on activation normal T cell expressed and secreted (RANTES) are elevated in the gastric mucosa by H. pylori. This study aimed to investigate the effects of Lactobacillus paracasei strain 06TCa19, a probiotic strain, on IL-8 and RANTES expression and production induced by H. pylori using human gastric epithelial cell lines. Strain 06TCa19 was shown to suppress H. pylori-mediated elevation of gene expression related to these chemokines in MKN45 cells. The strain also suppressed the increase in IL-8 and RANTES products induced by H. pylori in AGS cells as well as in MKN45 cells. In MKN45 cells inoculated with H. pylori, strain 06TCa19 was shown to downregulate the activation of NF-κB and p38 MAPK signaling pathways. Additionally, the level of the CagA virulence protein of H. pylori in the MKN45 cells and the number of viable H. pylori adhering to MKN45 cells decreased with the addition of strain 06TCa19. Moreover, the strain 06TCa19 notably increased lactic acid in the supernatant of MKN45 cells. Thus, lactic acid released from strain 06TCa19 might have inhibited the adhesion of H. pylori to MKN45 cells and prevented the insertion of H. pylori CagA into the cells, and elevation of IL-8 and RANTES genes and proteins might be suppressed by downregulating the NF-κB and p38 MAPK pathways. Therefore, use of strain 06TCa19 may prevent H. pylori-associated gastric inflammation.

Relationship between the Structure of α, β-Unsaturated Carbonyl Compounds and Bio-antimutagenic Activity against UV-Induced Escherichia coli WP2 B/r Trp^-

Relationship between the Structure of (α,βUnsaturated Carbonyl Compounds and Bioantimutagenic Activity against UV-Induced Escherichia coli WP2 B/r Trp Ryozo Iriye, Kohji Furukawa & Masahiko Takeshita To cite this article: Ryozo Iriye, Kohji Furukawa & Masahiko Takeshita (1992) Relationship between the Structure of (α,β-Unsaturated Carbonyl Compounds and Bio-antimutagenic Activity against UV-Induced Escherichia coli WP2 B/r Trp, Bioscience, Biotechnology, and Biochemistry, 56:3, 512-513, DOI: 10.1271/bbb.56.512 To link to this article: http://dx.doi.org/10.1271/bbb.56.512