title: Anti-HIV and immunomodulation activities of cacao mass lignin-carbohydrate complex.
abstract: BACKGROUND
  Recently, a prominent antiviral and macrophage stimulatory activity of cacao lignin-carbohydrate complex (LCC) has been reported. However, the solubility and sterility of LCC have not been considered yet. In the present study, complete solubilisation and sterilisation was achieved by autoclaving under mild alkaline conditions and the previously reported biological activities were re-examined.
  
  
  METHODS
  LCCs were obtained by 1% NaOH extraction and acid precipitation, and a repeated extraction-precipitation cycle. Nitric oxide (NO) and cytokine productions were assayed by the Griess method and ELISA, respectively. Inducible NO synthase (iNOS) expression was determined by Western blot analysis. Superoxide anion, hydroxyl radical and nitric oxide radical-scavenging activity was determined by ESR spectroscopy.
  
  
  RESULTS
  Cacao mass LCC showed reproducibly higher anti-HIV activity than cacao husk LCC. Cacao mass LCC, up to 62.5 μg/ml, did not stimulate mouse macrophage-like cells (RAW264.7 and J774.1) to produce NO, nor did it induce iNOS protein, in contrast to lipopolysaccharide (LPS). Cacao mass LCC and LPS synergistically stimulated iNOS protein expression, suggesting a different point of action. Cacao mass LCC induced tumour necrosis factor-α production markedly less than LPS, and did not induce interleukin-1β, interferon-α or interferon-γ. ESR spectroscopy showed that cacao mass LCC, but not LPS, scavenged NO produced from NOC-7.
  
  
  CONCLUSIONS
  This study demonstrated several new biological activities of LCCs distinct from LPS and further confirmed the promising antiviral and immunomodulating activities of LCCs.
authors.affiliation: Division of Pharmacology, Department of Diagnostic and Therapeutic Sciences, Meikai University School of Dentistry, Sakado, Saitama 350-0283, Japan. sakagami@dent.meikai.ac.jp
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sakagami
authors.forename: Michiyo
authors.initials: M
authors.lastname: Kawano
authors.forename: May Maw
authors.initials: MM
authors.lastname: Thet
authors.forename: Ken
authors.initials: K
authors.lastname: Hashimoto
authors.forename: Kazue
authors.initials: K
authors.lastname: Satoh
authors.forename: Taisei
authors.initials: T
authors.lastname: Kanamoto
authors.forename: Shigemi
authors.initials: S
authors.lastname: Terakubo
authors.forename: Hideki
authors.initials: H
authors.lastname: Nakashima
authors.forename: Yuji
authors.initials: Y
authors.lastname: Haishima
authors.forename: Yuuichi
authors.initials: Y
authors.lastname: Maeda
authors.forename: Koji
authors.initials: K
authors.lastname: Sakurai
mesh_terms: D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002099:Cacao / Q000737:chemistry*; D002241:Carbohydrates / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004578:Electron Spin Resonance Spectroscopy; D004797:Enzyme-Linked Immunosorbent Assay; D015497:HIV-1 / Q000187:drug effects / Q000254:growth & development; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology*; D008031:Lignin / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D010936:Plant Extracts / Q000494:pharmacology*; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology / Q000821:virology; D013481:Superoxides / Q000378:metabolism
chemical_list: D019380:Anti-HIV Agents; D002241:Carbohydrates; D016207:Cytokines; D007155:Immunologic Factors; D008070:Lipopolysaccharides; D010936:Plant Extracts; D013481:Superoxides; D009569:Nitric Oxide; D008031:Lignin; D052247:Nitric Oxide Synthase Type II