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Makoto Sunagawa - One of the best experts on this subject based on the ideXlab platform.

  • arabino mycolates derived from Cell Wall Skeleton of mycobacterium bovis bcg as a prominent structure for recognition by host immunity
    Drug discoveries and therapeutics, 2011
    Co-Authors: Masanori Miyauchi, Masashi Murata, Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Makoto Sunagawa, Keiko Shibuya, Erina Kogayamakawa, Yasuo Kashiwazaki
    Abstract:

    Arabino-mycolates are components of the Cell-Wall Skeleton of Mycobacterium bovis BCG (BCG-CWS). It is known that synthesized arabino- mycolates induce the production of tumor necrosis factor alpha (TNF-α) in murine macrophage Cell lines at an intensity similar to that of BCG-CWS. However the immunological activity of natural arabino-mycolates isolated from BCG has not been investigated, probably due to the complexity of the molecule. In this paper, we investigated the immunostimulatory activity of arabino-mycolates isolated from BCG-CWS by acid hydrolysis. Arabino-mycolates obtained by acid hydrolysis from the originally prepared CWS (SMP-105) of M. bovis BCG Tokyo 172 strain consisted mainly of mono-arabinose mono-mycolate, penta- arabinose tetra-mycolate and hexa-arabinose tetra- mycolate fractions. Arabino-mycolates significantly induced TNF-α production with an intensity comparable to that of CWS and enhanced delayed type hypersensitivity (DTH) reactions against inactivated tumor Cells. Arabino-mycolates-induced TNF-α production was completely dependent on TLR2 and MyD88 pathways. These findings indicate that isolated natural arabino-mycolates possess potent adjuvant immunostimulatory activity.

  • isolation and identification of arabinose mycolates of Cell Wall Skeleton cws derived from mycobacterium bovis bcg tokyo 172 smp 105
    Journal of Microbiological Methods, 2010
    Co-Authors: Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Makoto Sunagawa
    Abstract:

    A unique hydrolysis method using a two-layer solution, consisting of diluted hydrochloric acid and toluene was developed to isolate whole arabinose mycolates from the Cell Wall Skeleton of Mycobacterium bovis BCG Tokyo 172 (SMP-105) in order to reveal its pivotal role in enhancing immune responses against tumors.

  • separation and molecular characterization of mycolic acid from the Cell Wall Skeleton of mycobacterium bovis bcg tokyo 172 smp 105 and bcg substrains by normal phase high performance liquid chromatography and liquid chromatography mass spectrometry
    Journal of Microbiological Methods, 2009
    Co-Authors: Yuko Uenishi, Ikuya Yano, Takemasa Takii, Makoto Sunagawa
    Abstract:

    Since mycolic acids, the most characteristic major lipid component in mycobacterial Cell envelopes, play pivotal roles in the Cell surface-based host immune responses, normal-phase HPLC has been developed to quantify and identify mycolic acids of the Cell Wall Skeleton from Mycobacterium bovis BCG Tokyo 172 (SMP-105).

  • morphological study on mycobacterium bovis bcg tokyo 172 Cell Wall Skeleton smp 105
    Journal of Microbiological Methods, 2009
    Co-Authors: Yuko Uenishi, Keigo Kawabe, Takehiko Nomura, Masuyo Nakai, Makoto Sunagawa
    Abstract:

    Mycobacterial Cell Wall consists of rigid Cell Wall Skeleton (CWS), a mycoloyl arabinogalactan peptidiglycan complex, in which mycoloyl structure varies by the mycobacterial species diversely, whereas the arabinogalactan peptidoglycan structure is consistent comparatively. The CWS of Mycobacterium bovis BCG has long been expected as a potent adjuvant for immunotherapy of malignant tumor. Although the chemical structure of CWS has been established in the last few decades, the physicochemical properties of CWS having highly amphipathic miCelle structure with very long mycoloyl and carbohydrate chains are not unveiled. In this study, the ultrastructure of CWS of M. bovis BCG Tokyo 172 (SMP-105), suspended in several solvents with different polarity, was investigated with a particle size analyzer, a transmission electron microscope (TEM) and other techniques. As a result, the particle size was about 4.7 to 67.8 microm in physiological saline, but it became smaller and more compact when suspended in hydrophobic solvents. TEM images showed two different morphological forms distinctively: double folded sheet structure in hydrophilic conditions and multilayered rolled sheet structure in hydrophobic conditions. These studies have revealed characteristic surface features of SMP-105, the hydrophobic moiety occupying dominant space and the hydrophilic moiety smaller space, respectively, which may lead to the acceleration of immunological studies on this product.

  • comprehensive analysis of mycolic acid subclass and molecular species composition of mycobacterium bovis bcg tokyo 172 Cell Wall Skeleton smp 105
    Journal of Microbiological Methods, 2008
    Co-Authors: Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Yukiko Fujita, Makoto Sunagawa
    Abstract:

    The mycobacterial Cell envelope consists of a characteristic Cell Wall Skeleton (CWS), a mycoloyl arabinogalactan peptidoglycan complex, and related hydrophobic components that contribute to the Cell surface properties. Since mycolic acids have recently been reported to play crucial roles in host immune response, detailed molecular characterization of mycolic acid subclasses and sub-subclasses of CWS from Mycobacterium bovis BCG Tokyo 172 (SMP-105) was performed. Mycolic acids were liberated by alkali hydrolysis from SMP-105, and their methyl esters were separated by silica gel TLC into three subclasses: alpha-, methoxy-, and keto-mycolates. Each mycolate subclass was further separated by silver nitrate (AgNO(3))-coated silica gel TLC into sub-subclasses. Molecular weights of individual mycolic acid were determined by MALDI-TOF mass spectrometry. alpha-Mycolates were sub-grouped into cis, cis-dicyclopropanoic (alpha1), and cis-monocyclopropanoic-cis-monoenoic (alpha2) series; methoxy-mycolates were sub-grouped into cis-monocyclopropanoic (m1), trans-monocyclopropanoic (m2), trans-monoenoic (m3), cis-monocyclopropanoic-trans-monoenoic (m4), cis-monoenoic (m5), and cis-monocyclopropanoic-cis-monoenoic (m6) series; and keto-mycolates were sub-grouped into cis-monocyclopropanoic (k1), trans-monocyclopropanoic (k2), trans-monoenoic (k3), cis-monoenoic (k4), and cis-monocyclopropanoic-cis-monoenoic (k5) series. The position of each functional group, including cyclopropane rings and methoxy and keto groups, was determined by analysis of the meromycolates with fast atom bombardment (FAB) mass spectrometry and FAB mass-mass spectrometry, and the cis/trans ratio of cyclopropane rings and double bonds were determined by NMR analysis of methyl mycolates. Mycolic acid subclass and molecular species composition of SMP-105 showed characteristic features including newly-identified cis-monocyclopropanoic-trans-monoenoic mycolic acid (m4).

Yuko Uenishi - One of the best experts on this subject based on the ideXlab platform.

  • arabino mycolates derived from Cell Wall Skeleton of mycobacterium bovis bcg as a prominent structure for recognition by host immunity
    Drug discoveries and therapeutics, 2011
    Co-Authors: Masanori Miyauchi, Masashi Murata, Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Makoto Sunagawa, Keiko Shibuya, Erina Kogayamakawa, Yasuo Kashiwazaki
    Abstract:

    Arabino-mycolates are components of the Cell-Wall Skeleton of Mycobacterium bovis BCG (BCG-CWS). It is known that synthesized arabino- mycolates induce the production of tumor necrosis factor alpha (TNF-α) in murine macrophage Cell lines at an intensity similar to that of BCG-CWS. However the immunological activity of natural arabino-mycolates isolated from BCG has not been investigated, probably due to the complexity of the molecule. In this paper, we investigated the immunostimulatory activity of arabino-mycolates isolated from BCG-CWS by acid hydrolysis. Arabino-mycolates obtained by acid hydrolysis from the originally prepared CWS (SMP-105) of M. bovis BCG Tokyo 172 strain consisted mainly of mono-arabinose mono-mycolate, penta- arabinose tetra-mycolate and hexa-arabinose tetra- mycolate fractions. Arabino-mycolates significantly induced TNF-α production with an intensity comparable to that of CWS and enhanced delayed type hypersensitivity (DTH) reactions against inactivated tumor Cells. Arabino-mycolates-induced TNF-α production was completely dependent on TLR2 and MyD88 pathways. These findings indicate that isolated natural arabino-mycolates possess potent adjuvant immunostimulatory activity.

  • isolation and identification of arabinose mycolates of Cell Wall Skeleton cws derived from mycobacterium bovis bcg tokyo 172 smp 105
    Journal of Microbiological Methods, 2010
    Co-Authors: Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Makoto Sunagawa
    Abstract:

    A unique hydrolysis method using a two-layer solution, consisting of diluted hydrochloric acid and toluene was developed to isolate whole arabinose mycolates from the Cell Wall Skeleton of Mycobacterium bovis BCG Tokyo 172 (SMP-105) in order to reveal its pivotal role in enhancing immune responses against tumors.

  • separation and molecular characterization of mycolic acid from the Cell Wall Skeleton of mycobacterium bovis bcg tokyo 172 smp 105 and bcg substrains by normal phase high performance liquid chromatography and liquid chromatography mass spectrometry
    Journal of Microbiological Methods, 2009
    Co-Authors: Yuko Uenishi, Ikuya Yano, Takemasa Takii, Makoto Sunagawa
    Abstract:

    Since mycolic acids, the most characteristic major lipid component in mycobacterial Cell envelopes, play pivotal roles in the Cell surface-based host immune responses, normal-phase HPLC has been developed to quantify and identify mycolic acids of the Cell Wall Skeleton from Mycobacterium bovis BCG Tokyo 172 (SMP-105).

  • morphological study on mycobacterium bovis bcg tokyo 172 Cell Wall Skeleton smp 105
    Journal of Microbiological Methods, 2009
    Co-Authors: Yuko Uenishi, Keigo Kawabe, Takehiko Nomura, Masuyo Nakai, Makoto Sunagawa
    Abstract:

    Mycobacterial Cell Wall consists of rigid Cell Wall Skeleton (CWS), a mycoloyl arabinogalactan peptidiglycan complex, in which mycoloyl structure varies by the mycobacterial species diversely, whereas the arabinogalactan peptidoglycan structure is consistent comparatively. The CWS of Mycobacterium bovis BCG has long been expected as a potent adjuvant for immunotherapy of malignant tumor. Although the chemical structure of CWS has been established in the last few decades, the physicochemical properties of CWS having highly amphipathic miCelle structure with very long mycoloyl and carbohydrate chains are not unveiled. In this study, the ultrastructure of CWS of M. bovis BCG Tokyo 172 (SMP-105), suspended in several solvents with different polarity, was investigated with a particle size analyzer, a transmission electron microscope (TEM) and other techniques. As a result, the particle size was about 4.7 to 67.8 microm in physiological saline, but it became smaller and more compact when suspended in hydrophobic solvents. TEM images showed two different morphological forms distinctively: double folded sheet structure in hydrophilic conditions and multilayered rolled sheet structure in hydrophobic conditions. These studies have revealed characteristic surface features of SMP-105, the hydrophobic moiety occupying dominant space and the hydrophilic moiety smaller space, respectively, which may lead to the acceleration of immunological studies on this product.

  • comprehensive analysis of mycolic acid subclass and molecular species composition of mycobacterium bovis bcg tokyo 172 Cell Wall Skeleton smp 105
    Journal of Microbiological Methods, 2008
    Co-Authors: Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Yukiko Fujita, Makoto Sunagawa
    Abstract:

    The mycobacterial Cell envelope consists of a characteristic Cell Wall Skeleton (CWS), a mycoloyl arabinogalactan peptidoglycan complex, and related hydrophobic components that contribute to the Cell surface properties. Since mycolic acids have recently been reported to play crucial roles in host immune response, detailed molecular characterization of mycolic acid subclasses and sub-subclasses of CWS from Mycobacterium bovis BCG Tokyo 172 (SMP-105) was performed. Mycolic acids were liberated by alkali hydrolysis from SMP-105, and their methyl esters were separated by silica gel TLC into three subclasses: alpha-, methoxy-, and keto-mycolates. Each mycolate subclass was further separated by silver nitrate (AgNO(3))-coated silica gel TLC into sub-subclasses. Molecular weights of individual mycolic acid were determined by MALDI-TOF mass spectrometry. alpha-Mycolates were sub-grouped into cis, cis-dicyclopropanoic (alpha1), and cis-monocyclopropanoic-cis-monoenoic (alpha2) series; methoxy-mycolates were sub-grouped into cis-monocyclopropanoic (m1), trans-monocyclopropanoic (m2), trans-monoenoic (m3), cis-monocyclopropanoic-trans-monoenoic (m4), cis-monoenoic (m5), and cis-monocyclopropanoic-cis-monoenoic (m6) series; and keto-mycolates were sub-grouped into cis-monocyclopropanoic (k1), trans-monocyclopropanoic (k2), trans-monoenoic (k3), cis-monoenoic (k4), and cis-monocyclopropanoic-cis-monoenoic (k5) series. The position of each functional group, including cyclopropane rings and methoxy and keto groups, was determined by analysis of the meromycolates with fast atom bombardment (FAB) mass spectrometry and FAB mass-mass spectrometry, and the cis/trans ratio of cyclopropane rings and double bonds were determined by NMR analysis of methyl mycolates. Mycolic acid subclass and molecular species composition of SMP-105 showed characteristic features including newly-identified cis-monocyclopropanoic-trans-monoenoic mycolic acid (m4).

Tsukasa Seya - One of the best experts on this subject based on the ideXlab platform.

  • a possible abscopal effect of post irradiation immunotherapy in two patients with metastatic lung tumors
    International Cancer Conference Journal, 2014
    Co-Authors: Ken Kodama, Masahiko Higashiyama, Jiro Okami, Takashi Akazawa, Toshiteru Tokunaga, Norimitsu Inoue, Tsukasa Seya
    Abstract:

    As well as its local effects, radiotherapy leads to the delayed regression of distant non-irradiated lesions. These abscopal effects are most likely mediated by the innate immune system. Patient 1, a 74-year-old male, had concomitant left supraclavicular lymph node metastases and multiple lung metastases 2 years after complete resection of pathological stage IIA (T1bN1M0) lung adenocarcinoma. He received radiation therapy (RT) of 58 Gy for the supraclavicular lymph node metastases and then innate immunotherapy using the Cell Wall Skeleton of Mycobacterium bovis bacillus Calmette–Guerin (BCG-CWS). Three months after the RT and 2 months after the immunotherapy, all lung metastases disappeared on computed tomography scans. Patient 2, a 40-year-old female, underwent stereotactic body RT (SBRT) for metastasis from a deep-seated urothelial carcinoma in the right upper lobe of the lung. Twenty-one months after the SBRT, we started administration of BCG-CWS to two new lesions that had appeared in the left lung. As a result, after 3 months, the lesions completely disappeared. Complete response was maintained for more than 1 year in both patients. We believe that an optimal combination of RT and immunotherapy will elicit abscopal effects that can be employed to attain a systematically achievable, rather than anecdotal, therapeutic goal.

  • innate immune therapy with a bacillus calmette guerin Cell Wall Skeleton after radical surgery for non small Cell lung cancer a case control study
    Surgery Today, 2009
    Co-Authors: Ken Kodama, Misako Matsumoto, Tsukasa Seya, Masahiko Higashiyama, Koji Takami, Kazuyuki Oda, Jiro Okami, Jun Maeda, Takashi Akazawa, Mariko Wada
    Abstract:

    We investigated whether adjuvant immunotherapy with Bacillus Calmette-Guerin (BCG) Cell Wall Skeleton (CWS) and surgical resection was better than resection, with or without other adjuvant therapy, for patients with non-small Cell lung cancer (NSCLC). The case group comprised 71 patients who underwent radical surgery for NSCLC, followed by BCG-CWS immunotherapy, with follow-up data available. The case-control study was designed with one control selected for each case-group patient. Each control was matched by pathological stage and year of birth (±5 years). BCG-CWS 200 μg was inoculated intracutaneously in the upper arm four times per week (sensitization phase); then at 4-week intervals (therapeutic phase). The case-group patients received 45 ± 22.6 (average ± SD) cycles of BCG-CWS inoculation. Overall 5-year and 10-year survival rates were 71% and 61% for the case-group patients, and 63% and 43% for the control-group patients. The survival rate of the case group was better than that of the control group (not significant; P = 0.114). The same trend was seen in the patients with stage III or N+ NSCLC (not significant; P = 0.114, P = 0.168). There were no life-threatening adverse events. BCG-CWS immunotherapy seemed to improve survival after resection of NSCLC, especially locally advanced NSCLC. Moreover, this immunotherapy did not compromise quality of life during treatment.

  • gene inducing program of human dendritic Cells in response to bcg Cell Wall Skeleton cws which reflects adjuvancy required for tumor immunotherapy
    Immunology Letters, 2005
    Co-Authors: Kazuo Ishii, Misako Matsumoto, Yasuo Kashiwazaki, Mitsue Kuritataniguchi, Mikio Aoki, Toru Kimura, Tsukasa Seya
    Abstract:

    Adjuvants induce the expression of a number of genes in dendritic Cells (DCs), which facilitate effective antigen-presentation and cytokine/chemokine liberation. It has been accepted that the toll-like receptor (TLR) family governs the adjuvant activity in DCs. An adjuvant with a long history is mycobacteria in an oil-in-water emulsion, namely Freund's complete adjuvant. Since the active center for the adjuvancy in mycobacteria is the Cell-Wall Skeleton (CWS), we used the bacillus Calmette-Guerin Cell-Wall Skeleton (BCG-CWS) to test DC maturation by GeneChip analysis. We identified the genes supporting an efficient DC response and output. Approximately 2000 genes were up-regulated by BCG-CWS stimulation. BCG-CWS-, peptidoglycan (PGN)- and lipopolysaccharide (LPS)-stimulation generally up-regulated some gene clusters including genes for inflammatory cytokines (TNF, IL1alpha, IL1beta, IL6, IL12 p40, IL23 p19, etc.), chemokines (CCL20, IL8, etc.), Cell adhesion molecules (ICAM-1, etc.), apoptosis-related proteins (GADD45B, BCL2A1, etc.), metabolic enzymes (PTGS2, SOD2, etc.) and misCellaneous proteins (EHD1, TNFAIP6, etc.). LPS-stimulation, but not BCG-CWS- or PGN-stimulation, up-regulated the interferon-inducible antiviral proteins, including IFIT1, IFIT2, IFIT4, CXCL10, ISG15, OASL, IFITM1 and MX1. We also found that the BCG-CWS- or PGN-stimulation up-regulated CXCL5, MMP1, etc. We discussed their properties in association with TLRs and recently discovered TLR adapters.

  • structural functional relationship of pathogen associated molecular patterns lessons from bcg Cell Wall Skeleton and mycoplasma lipoprotein m161ag
    Microbes and Infection, 2002
    Co-Authors: Tsukasa Seya, Nasim A Begum, Ichiro Azuma, Shoutaro Tsuji, Misako Matsumoto, Kumao Toyoshima
    Abstract:

    The innate immune system senses microbial components by signaling receptors and induces phagocytosis by uptake receptors. The Toll-like receptor represents the signaling receptors that cause maturation of dendritic Cells, while phagocytosis is supported by other receptor families. We identify the structural signatures of microbial components recognized by these receptors to establish the two-receptor hypothesis in innate immunity.

  • activation of the human innate immune system by spirulina augmentation of interferon production and nk cytotoxicity by oral administration of hot water extract of spirulina platensis
    International Immunopharmacology, 2002
    Co-Authors: Tomohiro Hirahashi, Misako Matsumoto, Kaoru Hazeki, Yoshiko Saeki, Tsukasa Seya
    Abstract:

    Spirulina platensis is a cyanobacterial species that is surmised to potentiate the immune system leading to suppression of cancer development and viral infection. Here, we identified the molecular mechanism of the human immune potentiating capacity of Spirulina by analyzing blood Cells of volunteers with pre and post oral administration of hot water extract of Spirulina. NK functions represented by IFN gamma production and cytolysis were enhanced after administration of Spirulina in >50% subjects. IFN gamma was produced in an IL-12/IL-18-dependent fashion. In vitro stimulation of blood Cells with BCG Cell Wall Skeleton (CWS) allowed more potent IL-12 p40 production in Cells from volunteers given Spirulina than in Cells without pre-exposure to Spirulina. As BCG-CWS serves as a ligand for Toll-like receptor (TLR) 2 and 4 to raise the maturation stage of monocytes/macrophages, Spirulina may be involved in the signaling responses through Toll in blood Cells even when orally administered. These observations indicated that in humans Spirulina acts directly on myeloid lineages and either directly or indirectly on NK Cells. The presence of co-operative IL-12 and IL-18 is critically important for NK-mediated IFN gamma production.

Ikuya Yano - One of the best experts on this subject based on the ideXlab platform.

  • antitumor effect of cationic liposome incorporating keto mycolic acid from mycobacterium bovis bacillus calmette guerin in mouse model
    Journal of Clinical Oncology, 2018
    Co-Authors: Takayuki Yoshino, Ikuya Yano, Hideyuki Akaza, Jun Miyazaki, Takahiro Kojima, Shuya Kandori, Hiroyuki Nishiyama
    Abstract:

    461Background: Intravesical instillation of live bacillus Calmette-Guerin (BCG) is an established immunotherapy for non-muscle invasive bladder cancer. However, development of non-infectious agents has been expected, because there are several problems due to instillation of live bacteria. BCG Cell Wall Skeleton is known to stimulate host immune system, but the mechanism is still unknown. Mycolic acid (MA), which consist of three subclasses (α, keto, methoxy), is the most abundant lipid component of BCG Cell Wall and is expected to be one of essential active components. Objective: In order to identify active component from BCG Cell Wall and develop non-infectious BCG-derived immunotherapy, anti-tumor activities of cationic liposomes incorporating each subclass of MA were assessed using mouse syngeneic graft model. Methods: Heat killed packed Cells of M. bovis BCG Tokyo 172 were fully hydrolyzed and MA was obtained. MA was separated to three subclasses by thin layer chromatography. Cationic and hydrophilic ...

  • the liposome incorporating Cell Wall Skeleton of mycobacterium bovis bacillus calmette guein can directly enhance the susceptibility of cancer Cells to lymphokine activated killer Cells through up regulation of natural killer group 2 member d ligands
    BJUI, 2011
    Co-Authors: Jun Miyazaki, Ikuya Yano, Koji Kawai, Takahiro Kojima, Takehiro Oikawa, Akira Joraku, Toru Shimazui, Akihiro Nakaya, Takashi Nakamura, Hideyoshi Harashima
    Abstract:

    OBJECTIVE • To conduct a preclinical evaluation of the ability of natural killer Cells to cytolyze bladder cancer Cells that were modified to show enhanced expression of natural-killer group 2, member D (NKG2D) ligands by R8-liposome-bacillus Calmette-Guein (BCG)-Cell Wall Skeleton (CWS) treatment. MATERIALS AND METHODS • The T24 Cells and RT-112 Cells were co-cultured with R8-liposome-BCG-CWS and BCG for 2, 4, or 6 h, and then the surface expression of NKG2D ligands was analyzed using TaqMan real-time quantitative RT-PCR. • Peripheral blood mononuclear Cells were obtained with a conventional preparation kit, and then lymphokine-activated killer (LAK) Cells were generated from these purified peripheral blood mononuclear Cells via interleukin-2 stimulation. • The anti-tumour effect of LAK Cells against untreated and R8-liposome-BCG-CWS co-cultured with Cells of the human bladder cancer Cell lines T24 and RT-112 was analyzed using the cytotoxic WST-8 assay method at 4 h of culture at various effector/target (E : T) ratios. RESULTS • Major histocompatibility complex class I-related chain B (MICB) expression was increased ≈1.5-fold on T24 Cells and RT-112 Cells with BCG. • UL-16-binding protein (ULBP) 1 expression was also increased ≈1.5-fold on T24 Cells and RT-112 Cells with BCG. R8-liposome-BCG-CWS increased the surface expression of MICB 2.2-fold on T24 Cells but did not increase it significantly on RT-112 Cells. • ULBP1 expression was increased ≈2.2-fold on RT-112 Cells, although no differences were observed between the expression of ULBP2 and 3 with R8-liposome-BCG-CWS. • T24 Cells that were co-cultured with R8-liposome-BCG-CWS showed an ≈1.3-fold increase in sensitivity to cytolysis by LAK Cells at an E : T ratio of 4 and RT-112 Cells showed an ≈1.4-fold increase at an E : T ratio of 2. CONCLUSIONS • In the present study, the induction of surface NKG2D ligands by R8-liposome-BCG-CWS rendered cancer Cells more susceptible to cytolysis by LAK Cells. • T24 Cells and RT-112 Cells, even when cultured singly in the absence of immune Cells, can directly respond to R8-liposome-BCG-CWS. • The results obtained in the present study may therefore indicate a novel adoptive immunotherapy against bladder cancers.

  • arabino mycolates derived from Cell Wall Skeleton of mycobacterium bovis bcg as a prominent structure for recognition by host immunity
    Drug discoveries and therapeutics, 2011
    Co-Authors: Masanori Miyauchi, Masashi Murata, Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Makoto Sunagawa, Keiko Shibuya, Erina Kogayamakawa, Yasuo Kashiwazaki
    Abstract:

    Arabino-mycolates are components of the Cell-Wall Skeleton of Mycobacterium bovis BCG (BCG-CWS). It is known that synthesized arabino- mycolates induce the production of tumor necrosis factor alpha (TNF-α) in murine macrophage Cell lines at an intensity similar to that of BCG-CWS. However the immunological activity of natural arabino-mycolates isolated from BCG has not been investigated, probably due to the complexity of the molecule. In this paper, we investigated the immunostimulatory activity of arabino-mycolates isolated from BCG-CWS by acid hydrolysis. Arabino-mycolates obtained by acid hydrolysis from the originally prepared CWS (SMP-105) of M. bovis BCG Tokyo 172 strain consisted mainly of mono-arabinose mono-mycolate, penta- arabinose tetra-mycolate and hexa-arabinose tetra- mycolate fractions. Arabino-mycolates significantly induced TNF-α production with an intensity comparable to that of CWS and enhanced delayed type hypersensitivity (DTH) reactions against inactivated tumor Cells. Arabino-mycolates-induced TNF-α production was completely dependent on TLR2 and MyD88 pathways. These findings indicate that isolated natural arabino-mycolates possess potent adjuvant immunostimulatory activity.

  • bacillus calmette guerin and bcg Cell Wall Skeleton suppressed viability of bladder cancer Cells in vitro
    Anticancer Research, 2010
    Co-Authors: Tomoyuki Kato, Ikuya Yano, Vladimir Bilim, Kaori Yuuki, Sei Naito, Takuya Yamanobe, Akira Nagaoka, Hideyuki Akaza, Yoshihiko Tomita
    Abstract:

    Aim: Bacillus Calmette-Guerin (BCG) is one of therapeutic options for urothelial carcinoma (UC). The objectives of this study were to determine the direct effect of viable or heat-killed BCG and BCG Cell Wall Skeleton (BCG- CWS) on UC Cells in vitro. Materials and Methods: UC Cell lines were co-cultured with viable or heat-killed BCG Immunobladder ® (Tokyo 172 strain) and BCG-CWS. Viability of the Cells, apoptosis and BrdU incorporation were estimated. Results: BCG induced Cell growth retardation in highly malignant UC bearing integrin α5β1 (VLA5). VLA5- blocking antibody partially abrogated this effect. BCG treatment induced a modest increase in the sub-G 1 fraction of Cells and a decrease of BrdU incorporation. Cell growth retardation effect of viable BCG was reproduced by both heat-killed BCG and BCG-CWS. Conclusion: The results indicate that VLA5 may be a biomarker of UC with sensitivity to BCG. Moreover, BCG-CWS is a promising substance which might replace BCG, preventing life- threatening complications of viable BCG treatment. In 2009, an estimated 70,980 adults (52,810 men and 18,170 women) were diagnosed with bladder cancer in the United States, with 14,330 deaths (10,180 men and 4,150 women) (Cancer.Net by ASCO, http://www.cancer.net). Among men, bladder cancer is the fourth most common cancer and the eighth most common cause of cancer death (Cancer.Net by ASCO, http://www.cancer.net). Seventy-four percent of bladder cancer patients are diagnosed with noninvasive/superficial urothelial carcinoma (UC). The five-year relative survival rate is 93% for those patients (Cancer.Net by ASCO, http://www.cancer.net). Although superficial bladder cancer has favourable prognosis, it poses a substantial treatment challenge.

  • isolation and identification of arabinose mycolates of Cell Wall Skeleton cws derived from mycobacterium bovis bcg tokyo 172 smp 105
    Journal of Microbiological Methods, 2010
    Co-Authors: Yuko Uenishi, Naoto Kusunose, Ikuya Yano, Makoto Sunagawa
    Abstract:

    A unique hydrolysis method using a two-layer solution, consisting of diluted hydrochloric acid and toluene was developed to isolate whole arabinose mycolates from the Cell Wall Skeleton of Mycobacterium bovis BCG Tokyo 172 (SMP-105) in order to reveal its pivotal role in enhancing immune responses against tumors.

Akira Hayashi - One of the best experts on this subject based on the ideXlab platform.

  • immunotherapy of ovarian cancer with Cell Wall Skeleton of mycobacterium bovis bacillus calmette guerin effect of lymphadenectomy
    Cancer Science, 2009
    Co-Authors: Akira Hayashi, Yuichi Nishida, Saiji Yoshii, Seo Young Kim, Hirotsugu Uda, Toshimitsu Hamasaki
    Abstract:

    The significance of lymphadenectomy in surgery for various kinds of cancer has been widely debated, particularly in the gynecological field. The Cell Wall Skeleton of Mycobacterium bovis Bacillus Calmette-Guerin (BCG-CWS) has been used as an effective adjuvant for immunotherapy of a variety of cancer patients. Here we tested the immunological importance of lymph nodes in treatment of ovarian cancer patients with BCG-CWS. After surgical removal of tumors, 73 ovarian cancer patients were intracutaneously inoculated with BCG-CWS in the antigen-unloaded state in the upper arm at 4-week intervals at a dosage of 2–200 µg. Significant correlation of lymphadenectomy and reduced survival of patients was observed (stages I, II, III, IV; hazard ratio 2.38, 95% confidence interval 1.02–5.12, Cox regression model). Lymphadenectomy also compromised with induction of interferon-γ. In view of the importance of the role of lymph nodes in stimulation of Toll-like receptors by BCG-CWS, it is suggested that lymph nodes should be kept as much as possible to preserve the patient's immunity against cancer. Application of these results to surgery for other cancers should be considered. (Cancer Sci 2009; 100: 1991–1995)

  • maturation of human dendritic Cells by Cell Wall Skeleton of mycobacterium bovis bacillus calmette guerin involvement of toll like receptors
    Infection and Immunity, 2000
    Co-Authors: Shoutaro Tsuji, Ichiro Azuma, Kumao Toyoshima, Misako Matsumoto, Osamu Takeuchi, Shizuo Akira, Akira Hayashi, Tsukasa Seya
    Abstract:

    The constituents of mycobacteria are an effective immune adjuvant, as observed with complete Freund's adjuvant. In this study, we demonstrated that the Cell Wall Skeleton of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS), a purified noninfectious material consisting of peptidoglycan, arabinogalactan, and mycolic acids, induces maturation of human dendritic Cells (DC). Surface expression of CD40, CD80, CD83, and CD86 was increased by BCG-CWS on human immature DC, and the effect was similar to those of interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), heat-killed BCG, and viable BCG. BCG-CWS induced the secretion of TNF-alpha, IL-6, and IL-12 p40. CD83 expression was increased by a soluble factor secreted from BCG-CWS-treated DC and was completely inhibited by monoclonal antibodies against TNF-alpha. BCG-CWS-treated DC stimulated extensive allogeneic mixed lymphocyte reactions. The level of TNF-alpha secreted through BCG-CWS was partially suppressed in murine macrophages with no Toll-like receptor 2 (TLR 2) or TLR4 and was completely lost in TLR2 and TLR4 double-deficient macrophages. These results suggest that the BCG-CWS induces TNF-alpha secretion from DC via TLR2 and TLR4 and that the secreted TNF-alpha induces the maturation of DC per se.

  • bcg Cell Wall Skeleton completely cures the immunologically eligible acute leukemia patients
    Proceedings of the Japan Academy. Ser. B: Physical and Biological Sciences, 1999
    Co-Authors: Akira Hayashi, Hiroyuki Nakamura, Takuro Sugihara, Ichiro Azuma
    Abstract:

    Eight acute leukemia patients were treated with BCG-Cell Wall Skeleton (CWS) alone after the first complete remission (CR) induction with chemotherapy alone or chemotherapy followed by autologous peripheral blood stem Cell transplantation (PBSCT). Two patients, who achieved CR after one course of chemotherapy, were treated with BCG-CWS alone for about 8 years, one starting just after the first relapse and the other just after the first remission. They have been in CR state for more than two decades, without further relapse or other secondary cancers. Recently, their complete cure was confirmed; Wilms tumor (WT)-1 gene expression in their peripheral blood was less than 10-5. Among other 6 patients, who had repeated or intensive chemotherapy, 4 patients responded to immunotherapy with BCG-CWS alone are alive in CR for more than 1 year, but two died shortly thereafter. The most important factor influencing the above patients' fate might be their own immunity at the start of immunotherapy; this can be monitored with the interferon (IFN)-γ induction test. In view of above results, immunotherapy with BCG-CWS alone for acute leukemia patients should be used just after the first CR and before transplantation, because of its priority in such terms as simple procedure, high cost-performance, high quality of life, easy availability and no risk of tuberculosis.

  • immuno friendly use of bcg Cell Wall Skeleton remarkably improves the survival rate of various cancer patients
    Proceedings of the Japan Academy. Series B Physical and biological sciences, 1998
    Co-Authors: Akira Hayashi, Ichiro Azuma, Osamu Doi, Kumao Toyoshima
    Abstract:

    About 25 years ago, the study of cancer immunotherapy (IT) was done without paying due attention to patients' own immunogenicity, resulting in the failure in its clinical trials. This fact suggested that IT should be performed independently of other conventional chemo- and/or radiation therapies after removing as many tumor Cells as possible. Between 1975 and 1997, total 235 cancer patients have been treated by IT with BCG-Cell Wall Skeleton (CWS) alone, most just after surgical operations, and some after chemo- and/or radiation therapies. Appropriate dose (10 to 200μg) of BCG-CWS depending on the patients' biological responses, inoculated intracutaneously every 4 weeks, monitoring their immunogenicity with IFN-γ induction test. In this pilot study, clinical effect of the immuno-friendly use of BCG-CWS was shown most clearly in the effect of IFN-γ induction and cancer stages on the survival rate. In fact this therapy was really effective for various kinds of cancers and for the patients in Stage I-III, but not for those in Stage IV or in immunologically impaired state. Further in 84 postoperative primary lung cancer patients, especially in the group of 46 Stage III patients, the effect was statistically significant (χ2=8.546, p=0.0035). These clinical and statistical analyses suggest that the IT with BCG-CWS alone displays its effect on cancers attacking their lymphnode metastasis.

  • does the Cell Wall Skeleton from bacille calmette guerin directly induce interferon independent of interleukin 12
    Japanese Journal of Clinical Oncology, 1996
    Co-Authors: Akira Hayashi, Atsunari Noda
    Abstract:

    lnterleukin-12 (IL-12), known to be a strong inducer of interferon-7 (IFN-7), plays a vital role in activating the immune surveillance system against intraCellular pathogens and malignant tumors. The authors have found that cancer patients showing marked IFN-7 induction after inoculation with BCG-CWS (the Cell Wall Skeleton from Bacille Calmette-Guerin) have a good prognosis. The present study was undertaken to determine whether the level of IL-12 is increased prior to, or along with, IFN-7 induction in the serum of patients inoculated with BCG-CWS. Unexpectedly, we found no detectable amount of IL-12 in the serum throughout the entire time course. This suggests that a novel IFN-7 inducing factor (IGIF) or another unknown IFN-7 inducer may be working in place of IL-12 in the BCG-CWS system. (Jpn J Clin Oncol 26: 124-127, 1996)