1986;154:737. of rhamnogalacturonan core rich in neutral sugar chain, showed the potent mitogenic activity suggesting it to be an active site. Mitogenic activity of bupleuran 2IIc was reduced in the presence of antipolysaccharide antibody (antibupleuran 2IIc/PG-1-IgG), which recognizes the ramified region of bupleuran 2IIc as the antigenic epitope. Mitogenic activity of bupleuran 2IIc was also reduced by the addition of -d-Glcor -d-Glcand pharmacological activities in addition to the software for drug delivery.1 Therefore, it is possible to consider pectic polysaccharide, not only as one of the active ingredients of medicinal herbs, but also as medicines. The origins of L. (Japanese name = L. Bupleuran 2IIb and 2IIc consist of a galacturonan region, a ramified region composed of a rhamnogalacturonan core having neutral sugars side chains, and a rhamnogalacturonan II-like region;7 the ramified region has been considered to be an important part of the expression of these activities.2C4 Therefore, a polyclonal antibody (antibupleuran 2IIc/PG-1-IgG) against the ramified region (PG-1) of bupleuran 2IIc was prepared and applied for analysis of absorption and cells distribution of the polysaccharide after oral administration to mice.8 The effects suggested that a portion of bupleuran 2IIc was absorbed in the body YZ9 from your digestive organs, and then mobilized into liver.8 The polysaccharide was also detected in Peyers patches of the mucosa-associated lymphoid YZ9 cells (MALT) on intestine.8 Because most herbal medicines are taken orally, YZ9 it is assumed that some active ingredients in the medicines also transport to lymphoid cells to interact with lymphocytes. The major antigenic epitope against antibupleuran 2IIc/PG-1-IgG was characterized to be 6-linked galactosyl chains comprising terminal glucuronic acid (GlcA) or 4-L. on murine lymphocytes and (O127:B8) and HEPES were from Sigma (St. Louis, MO). Preparation of bupleuran 2IIc and subfractionsThe origins of L. were purchased from Uchida Wakanyaku Co. Ltd. (Tokyo, Japan). A voucher specimen was deposited in the herbarium of Oriental Medicine Research Center of the Kitasato Institute. Bupleurans 2IIb and 2IIc were purified from your pectic polysaccharide portion (BR-2) of the origins of by anion-exchange chromatography on diethylaminoethyl (DEAE)CSepharose CL-6B (Pharmacia Good Chemicals, Uppsala, Sweden) as explained previously.5Ramified region (PG-1; rhamnogalacturonan core possessing side chains rich in neutral sugars) and PG-3 (oligogalacturonide portion) were prepared from bupleuran 2IIc by endo-(14)–d-polygalacturonase digestion, as reported previously.5-d-Glcand -d-Glcfrom Acacia gum were prepared as reported by Tsumuraya was isolated from your partial acid hydrolysate of larch wood arabinogalactan.10 MediaRPMI-1640 medium was supplemented with penicillin (100 U/ml), streptomycin (10 g/ml) amphotericin B (025 g/ml) and 125 mm HEPES. Furthermore, this medium was regularly RBX1 supplemented with 5% heat-inactivated fetal calf serum (RPMI-1640CFCS). In all experiments, 5 10?5 m 2-mercaptoethanol (2-ME) was added to the medium. AntibodiesThe polyclonal antibody (antibupleuran 2IIc/PG-1-immunoglobulin G (IgG)) against the ramified region (PG-1) of bupleuran 2IIc was generated and purified as explained previously.8 Antibupleuran 2IIc/PG-1-IgG was purified by protein G-Sepharose and bupleuran 2IIc/PG-1 immobilized EAHCSepharose.8 Anti-bupleuran 2IIc/PG-1-F(ab)2 was prepared from antibupleuran 2IIc/PG-1-IgG by pepsin digestion. Dental administration YZ9 of sampleBR-2 (comprising bupleuran 2IIb and bupleuran 2IIc, 250 mg/kg) was dissolved in distilled water, and aliquots (02 ml) of the perfect solution is were given orally into mice. As the settings, the mice received distilled water only instead of the sample remedy. Cell preparation and cell cultureMice were killed by cervical amputation and their spleens were eliminated using aseptic techniques. The spleens were approved through a sterilized stainless sieve (150 mesh) to obtain a single-cell suspension. Erythrocytes in the cell suspension were destroyed by.