title: Immunomodulation by SanPharma fungal metabolic products.
abstract: OBJECTIVE
  In the present study, a series of fungal metabolite products from SanPharma (Dohren, Germany) were tested for effects on human peripheral blood leukocytes in vitro using standard immunologic methods.
  
  
  BACKGROUND
  Therapeutic strategies used in German biological medicine often include treatment (oral, nasal, rectal, topical, or injection) with fungal or bacterial products, also known as "isopathic remedies," of which some are limited to metabolic products, whereas others include microbial cell lysates and cell wall fragments as well. The SanPharma products are based on metabolites, and do not contain microbial cell wall compounds.
  
  
  METHODS
  Activation of natural killer (NK) cells was evaluated by cell surface immunostaining using CD3, CD56, CD69, and CD25 monoclonal antibodies. Production of interferon-gamma was evaluated by enzyme-linked immunoabsorbent assay (ELISA) on supernatants collected after 5 days' culture of peripheral blood mononuclear cells (PBMC). Direct mitogenic effect was assessed using the lipophilic membrane dye PHK26 (Sigma-Aldrich, St. Louis, MO) in a fluorescence-based proliferation assay, in which fluorescence intensity is reduced upon cell divisions. Cell viability upon exposure to fungal metabolites was assessed using propidium iodide staining and flow cytometry.
  
  
  RESULTS
  All fungal metabolite products specifically induced the expression of the CD69 marker on human CD3-negative, CD56-positive NK cells, but not CD3-positive T cells, in vitro, as shown by the induction of the CD69 marker on up to 50% of NK cells after 18 hours' culture with metabolites. Only one of the five metabolite products, Roqueforti, induced cyclooxygenase-2 (COX-2), indicating that nuclear factor-kappaB (NFkappaB)-mediated signaling may not have been involved in the NK activation by the other four products. The Notatum product reduced baseline levels of COX-2, indicating an anti-inflammatory effect. No evidence of toxic or mitogenic effects was found.
  
  
  CONCLUSIONS
  The fungal metabolite products from SanPharma specifically activate human NK cells in vitro.
authors.affiliation: Holger NIS, Inc., Port Dover, Ontario, Canada. Gitte@holgernis.com
authors.forename: Gitte S
authors.initials: GS
authors.lastname: Jensen
authors.forename: Aaron N
authors.initials: AN
authors.lastname: Hart
mesh_terms: D015703:Antigens, CD / Q000096:biosynthesis*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis; D017252:CD3 Complex / Q000096:biosynthesis; D019002:CD56 Antigen / Q000096:biosynthesis; D002457:Cell Extracts / Q000494:pharmacology*; D000529:Complementary Therapies / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005658:Fungi / Q000378:metabolism; D007155:Immunologic Factors / Q000494:pharmacology*; D016219:Immunotherapy, Adoptive; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000096:biosynthesis; D007694:Killer Cells, Natural / Q000276:immunology*; D037181:Lectins, C-Type; D008213:Lymphocyte Activation / Q000276:immunology*; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D019002:CD56 Antigen; D002457:Cell Extracts; D007155:Immunologic Factors; D007371:Interferon-gamma; D037181:Lectins, C-Type; D015375:Receptors, Interleukin-2; D000099192:CD69 Antigens