title: Evaluation of widely consumed botanicals as immunological adjuvants.
abstract: BACKGROUND
  Many widely used botanical medicines are claimed to be immune enhancers. Clear evidence of augmentation of immune responses in vivo is lacking in most cases. To select botanicals for further study based on immune enhancing activity, we study them here mixed with antigen and injected subcutaneously (s.c.). Globo H and GD3 are cell surface carbohydrates expressed on glycolipids or glycoproteins on the cell surface of many cancers. When conjugated to keyhole limpet hemocyanin (KLH), mixed with an immunological adjuvant and administered s.c. the magnitude of the antibody responses against globo H, GD3 and KLH depend largely on the potency of the adjuvant. We describe here the results obtained using this s.c. immunization model with seven botanicals purported to have immune stimulant effects.
  
  
  METHODS
  Groups of 5-10 mice were immunized with globo H-KLH or GD3-KLH mixed with botanical, saline or positive control immunological adjuvant, s.c. three times at 1 week intervals. Antibody responses were measured 1 and 2 weeks after the 3rd immunization. The following seven botanicals and fractions were tested: (1) H-48 (Honso USA Co.), (2) Coriolus versicolor raw water extract, purified polysaccharide-K (PSK) or purified polysaccharide-peptide (PSP) (Institute of Chinese Medicine (ICM)), (3) Maitake extract (Yukiguni Maitake Co. Ltd. and Tradeworks Group), (4) Echinacea lipophilic, neutral and acidic extracts (Gaia Herbs), (5) Astragalus water, 50% or 95% ethanol extracts (ICM), (6) Turmeric supercritical (SC) or hydro-ethanolic (HE) extracts (New Chapter) or 60% ethanol extract (ICM) and (7) yeast beta-glucan (Biotec Pharmacon). Purified saponin extract QS-21 (Antigenics) and semisynthetic saponin GPI-0100 (Advanced BioTherapies) were used as positive control adjuvants. Sera were analyzed by ELISA against synthetic globo H ceramide or GD3 and KLH.
  
  
  RESULTS
  Consistent significant adjuvant activity was observed after s.c. vaccination with the Coriolus extracts (especially PSK), a 95% ethanol extract of Astragalus and yeast beta-glucan, and (to a lesser extent) Maitake. Antibodies against KLH in all cases and against globo H in most cases were induced by these botanicals. Little or no adjuvant activity was demonstrated with H-48 or Echinacea extracts or the Astragalus water extract. Experiments with GD3-KLH as immunogen confirmed the adjuvant activity of the Coriolus, yeast beta-glucan and Astragalus extracts. While extraction with ethanol concentrated the active ingredients in Astragalus, it had no impact on Coriolus where the 90% ethanol precipitate and solute were equally active.
  
  
  CONCLUSIONS
  Some, but not all, botanicals purported to be immune stimulants had adjuvant activity in our model. PSK and Astragalus were surprisingly active and are being further fractionated to identify the most active adjuvant components.
authors.affiliation: Laboratory of Tumor Vaccinology, Melanoma and Sarcoma Service, Department of Medicine, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, United States. ragupatg@mskcc.org
authors.forename: Govind
authors.initials: G
authors.lastname: Ragupathi
authors.forename: K Simon
authors.initials: KS
authors.lastname: Yeung
authors.forename: Ping-Chung
authors.initials: PC
authors.lastname: Leung
authors.forename: Mavis
authors.initials: M
authors.lastname: Lee
authors.forename: Clara Bik San
authors.initials: CB
authors.lastname: Lau
authors.forename: Andrew
authors.initials: A
authors.lastname: Vickers
authors.forename: Chandra
authors.initials: C
authors.lastname: Hood
authors.forename: Gary
authors.initials: G
authors.lastname: Deng
authors.forename: Nai-Kong
authors.initials: NK
authors.lastname: Cheung
authors.forename: Barrie
authors.initials: B
authors.lastname: Cassileth
authors.forename: Philip
authors.initials: P
authors.lastname: Livingston
mesh_terms: D000276:Adjuvants, Immunologic / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D000912:Antibodies, Neoplasm / Q000097:blood*; D019496:Cancer Vaccines / Q000008:administration & dosage / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005658:Fungi / Q000737:chemistry*; D005732:Gangliosides / Q000276:immunology; D007117:Immunization, Secondary; D007279:Injections, Subcutaneous; D051379:Mice; D008810:Mice, Inbred C57BL; D010936:Plant Extracts / Q000494:pharmacology*; D010946:Plants, Medicinal / Q000737:chemistry*; D018074:Vaccines, Conjugate / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D000912:Antibodies, Neoplasm; D019496:Cancer Vaccines; C405428:GD3-lactone-KLH; D005732:Gangliosides; D010936:Plant Extracts; D018074:Vaccines, Conjugate; C119370:globo H-keyhole limpet hemocyanin QS-21 vaccine