title: Macrophages, rather than T and B cells are principal immunostimulatory target cells of Lycium barbarum L. polysaccharide LBPF4-OL.
abstract: OBJECTIVE
  Lycium barbarum L. is a renowned Yin strengthening agent in traditional Chinese medicine. Lycium barbarum L. polysaccharide-protein complex is well-known for its immunoregulatory and antitumor effects. LBPF4-OL is the glycan part of Lycium barbarum L. polysaccharide-protein complex fraction 4 (LBPF4). LBPF4-OL's active contribution in LBPF4 is still blank. In the study, we enrich the polysaccharide part of Lycium barbarum L. polysaccharide-protein complex, and investigate its immunostimulatory effects on mouse spleen cells, T cells, B cells and macrophages.
  
  
  METHODS
  Balb/C mice were used in vitro and in vivo studies. In in vitro study, lymphocyte proliferations were analyzed with (3)H-TdR incorporation method. Miltenyi MicroBeads were used in the purification of lymphocytes. Activation of T and B cells was analyzed by flow cytometry. In order to obtain the peritoneal macrophages, mice were injected i.p. with 1mL of sodium thioglycollate 3 days prior to killing. Spleen cells were stimulated with LBPF4-OL and cytokine concentrations in the supernatants were determined by multiplex bead analysis. In in vivo study, mice were injected i.p. with 1 mL of normal saline or 100 μg/mL LBPF4-OL daily for 6 days. Peritoneal macrophage functions were analyzed by enzyme-linked immunosorbent assay and flow cytometry assay.
  
  
  RESULTS
  Spleen cells and lymphocyte proliferation assay indicated that LBPF4-OL markedly induced the spleen cell proliferation, but could not induce proliferation of purified T and B lymphocytes. Further research revealed that B cell proliferation took place in the presence of activated macrophages or LPS. Multiplex bead analysis showed that LBPF4-OL can obviously induce IL-6, IL-8, IL-10 and TNF-α production of the spleen cells in a concentration-dependent manner. Flow cytometric analysis showed that LBPF4-OL (i.p.) prompts CD86 and MHC-II molecules expression on macrophages. ELISA assay showed that LBPF4-OL can greatly strengthen macrophage releasing of TNF-α and IL-1β.
  
  
  CONCLUSIONS
  These results suggested that glycan LBPF4-OL plays an important role in the immunopharmacological activity of Lycium barbarum L. polysaccharide-protein complex, and primary mouse macrophages, rather than T and B cells, are the principal target cells of it.
authors.affiliation: Beijing Institute of Pharmacology and Toxicology, Beijing, China.
authors.forename: Xiao Rui
authors.initials: XR
authors.lastname: Zhang
authors.forename: Wen Xia
authors.initials: WX
authors.lastname: Zhou
authors.forename: Yong Xiang
authors.initials: YX
authors.lastname: Zhang
authors.forename: Chun Hui
authors.initials: CH
authors.lastname: Qi
authors.forename: Huanga
authors.initials: H
authors.lastname: Yan
authors.forename: Zhong Fu
authors.initials: ZF
authors.lastname: Wang
authors.forename: Bo
authors.initials: B
authors.lastname: Wang
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects*; D051940:B7-2 Antigen / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005260:Female; D005638:Fruit; D008070:Lipopolysaccharides; D032305:Lycium / Q000737:chemistry*; D008213:Lymphocyte Activation / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D008285:Major Histocompatibility Complex / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D011134:Polysaccharides / Q000494:pharmacology*; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013601:T-Lymphocytes / Q000187:drug effects*
chemical_list: D000276:Adjuvants, Immunologic; D051940:B7-2 Antigen; D016207:Cytokines; D004365:Drugs, Chinese Herbal; D008070:Lipopolysaccharides; D011134:Polysaccharides; C500223:lycium barbarum polysaccharide