Protein-bound Polysaccharide Research Articles

Immunochemotherapy benefits in gastric cancer patients stratified by programmed death-1 ligand-1

BackgroundEffectiveness of protein-bound polysaccharide K (PSK) during adjuvant chemotherapy in gastric cancer patients expressing programmed death-1 ligand 1 (PD-L1) has not been investigated. Investigating this might help in triaging candidates eligible to immunochemotherapy. Materials and methodsIn total, 918 patients with stages II and III gastric cancer, undergoing curative gastrectomy, and receiving adjuvant chemotherapy were enrolled in a prospective database, and the patients were retrospectively reviewed. We classified those patients into four cohorts stratified by PD-L1 expression and PSK administration, namely PD-L1, PSK (−,+); PD-L1, PSK (−,−); PD-L1, PSK (+,+); and PD-L1, PSK (+,−). In addition, another independent cohort of 20 patients undergoing radical gastrectomy was prospectively recruited to check their immunological cells of sera before and 2 mo after PSK administration. ResultsPSK treatment was an independent prognostic factor for patient's overall survival (P = 0.020), whereas PD-L1 expression per se was not. Administration of PSK prolonged patient survival in stages IIIA and IIIB (P = 0.031) but not in stage II or stage IIIC. Patients with negative expression of PD-L1, treated with PSK had longer survival than those not treated with PSK (P = 0.033). PSK did not affect the survival of patients with positive expression of PD-L1, (P = 0.421). The percentages of natural killer and natural killer T (NKT) cells, but not Th1, Th17, Treg, or IFN-γ+/CD8+ T cells, were significantly increased in PD-L1 (−) patients treated with PSK. However, these findings were not evident in PD-L1 (+) patients. ConclusionsPSK treatment preferentially confers a survival gain for patients with stage IIIA/IIIB gastric cancer, especially in the PD-L1 (−) subpopulation.

Purification, characterization and bioactivity determination of a novel polysaccharide from pumpkin (Cucurbita moschata) seeds

The crude polysaccharides from pumpkin seeds were obtained with hot water extraction and ethanol precipitation. After purification by Q-Sepharose FF column and Sephadex G-100 gelfiltration, a protein-bound polysaccharide (PSP-I) was obtained and characterized. PSP-I was composed of mannose, glucose and galactose in a molar ratio of 1.0:12.9:5.3 with molecular weight of 21,100 g/mol. 1H NMR and 13C NMR spectra revealed that the backbone of PSP-I consisted of (1 → 4)-linked β-d-glucopyranosyl, (1 → 2)-linked α-d-galactose and (1 → 6)-linked α-d-mannopyranosyl. PSP-I contained 17 general amino acids and was rich in glutamic acid, alanine and glycine. The alkali sensitive O-glycoprotein was identified by β-elimination reaction. The antioxidative assays in vitro indicated that PSP-I showed the moderate scavenging activities on DPPH radical and hydroxyl radical. Moreover, PSP-I displayed the obvious inhibitory activity against α-amylase and strong antibacterial activity.

Fever-range hyperthermia inhibits cells immune response to protein-bound polysaccharides derived from Coriolus versicolor extract.

The aim of the study was to explore whether fever-range hyperthermia (FRH) might enhance the anticancer and immunoregulatory activities of protein-bound polysaccharides (PBP), a class of fungus derived immunomodifiers used in the cancer adjuvant therapy. Blood lymphocytes and breast cancer cells (MCF-7) were cultured at 39.5°C in humidified atmosphere containing 5% CO2 for 2h. After rested at 37°C for 6h, the cells were treated with PBP extract at 100- and 300μg/ml concentration. After indicated time, the proliferative response was analyzed and cytokine mRNA expression assessment was performed by qRT-PCR. In animal model, the FRH was induced by placing rats in the Homeothermic Controller with heating blanket. Animals were heated until Tb reached 39.5°C (±0.2°C) and were maintained at this temperature for 30min. The protein-bound polysaccharides solution was injected i.p. at a dose of 100 mg/kg 6h post FRH. Twenty four hours after treatment, the blood was collected and cytokines expression analysis were performed. The results have shown that fever-range hyperthermia has an inhibitory effect on PBP extract-induced proliferative response of blood lymphocytes, as well as IL-1β and IL-6 mRNA expression. Moreover, the temperature of 39.5°C blocks PBP-induced cytotoxicity against MCF-7 cells, which correlates with significant reduction in TNF-α level. Combined treatment of rats (FRH+PBP) results in decrease of IL-1β, IL-6 and TNF-α mRNA expression in peripheral blood mononuclear cells compared to cells derived from rats treated with protein-bound polysaccharides extract alone. This study demonstrates that fever-range temperature inhibits immunostimulatory as well as anticancer effects mediated by protein-bound polysaccharides.

Protein-bound polysaccharides from Coriolus versicolor attenuate LPS-induced synthesis of pro-inflammatory cytokines and stimulate PBMCs proliferation

Protein-bound polysaccharides (PBP) isolated from Coriolus versicolor (CV) are classified as biological response modifiers capable of exhibiting various biological activities, such as anti-tumour and immunopotentiating activity. Since we have found in vivo studies that the tested PBP induced prolongation of endotoxin fever in rats, the aim of the present study was to investigate the in vitro effect of the PBP on the production of pro-inflammatory cytokines by the lipolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs). The results showed that the PBP affect the immunomodulating properties of the LPS-treated PBMCs by the enhancement of mitogenic activity and attenuation of the LPS-induced production of interleukin (IL)-1β and IL-6. Moreover, the tested polysaccharides peptides themselves also exhibit immunomodulatory properties manifested in the increased cell proliferation and pro-inflammatory cytokine release from PBMCs. The effect of PBP on the both phenomena was time-dependent and occurred in the U-shaped dose response manner. These findings are significant when considering the use of commercially available PBP from CV extract by cancer patients suffering from immunodeficiency, who may experience microbial infections during therapy.

Metabolic mechanism of a polysaccharide from Schisandra chinensis to relieve chronic fatigue syndrome

Schisandra chinensis fruits are a famous traditional Chinese medicine to treat all kinds of fatigue. This study aimed to investigate the therapeutic effect and metabolic mechanism of a polysaccharide (SCP) from Schisandra chinensis fruits on chronic fatigue syndrome (CFS). SCP was isolated and the physicochemical properties were analyzed. A CFS model of rats was established and the urinary metabonomic studies were performed using gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) in combination with multivariate statistical analysis. The results showed that SCP is a protein-bound polysaccharide. The amino acid composition of SCP consisted of 12 amino acids. The growth and the behaviors of the rats in the CFS model group were worse than those in the control group and improved after SCP treatment. Analysis of the GC-TOF-MS revealed that twelve metabolites were significantly changed, and six metabolites were oppositely and significantly changed after the SCP treatment. The TCA cycle metabolic pathways and the alanine, aspartate and glutamate metabolism were identified as significant metabolic pathways involved with SCP. The therapeutic mechanism of SCP against CFS was partially due to the restoration of these disturbed pathways.

Polysaccharide peptides from Coriolus versicolor exert differential immunomodulatory effects on blood lymphocytes and breast cancer cell line MCF-7 in vitro

The protein-bound polysaccharides (PBP), isolated from Coriolus versicolor (CV) fungus, are considered as natural compounds with potential therapeutic applications. The immunopotentiating and antitumor activity of polysaccharopeptides has been previously examined, however similar findings could not be achieved. The source of PBP, variations in extraction process as well as environmental factors seems to affect the biological properties of these active CV components. Since further analysis are needed to draw more definite conclusion, the present study aimed to investigate the immunomodulatory properties of the PBP extract, isolated from commercially available capsules of C. versicolor. Our results revealed that the effect mediated by PBP extract depends on the target cells. We reported that the polysaccharopeptides induced a significant decrease in breast cancer MCF-7 cells growth, which was TNF-α-dependent phenomenon. Interestingly, the level of two others cytokines, IL-1β and IL-6 was not affected. On the other hand, in this study we noticed that protein-bound polysaccharides extracted from CV significantly augmented the proliferative response of blood lymphocytes in a time-dependent manner, which was associated with IL-6 and IL-1β mRNA upregulation. Moreover we found that the cells response to PBP stimuli might be inversely related to its concentration.

ANTIOXIDANT ACTIVITY EVALUATION OF METHANOLIC EXTRACT AND CRUDE POLYSACCHARIDES FROM Plumeria alba L. leaves

Methanolic (MeOH) extract and crude polysaccharides of Plumeria alba leaves were evaluated for in vitro antioxidant activity. HPLC analysis for the MeOH extract revealed the presence of ten flavonoids and nine phenolic compounds with higher antioxidant capacity. Total phenolic compounds and total flavonoids were determined. The monosaccharide composition was performed on a Dionex system (Dx-120) ion chromatography. The detected polysaccharides are heteropolysaccharides, glucose and mannose were occupied the highest level followed by galactose and rhamnose. Additionally, Plumeria alba polysaccharides (PAPs) consisted of the protein-bound polysaccharide. Furthermore, 14 amino acids (Asp, Thr, Glu, Ser, Ala, Gly, Arg, Tyr, Cys, Ile, Lys, Val, Pro and Phe) were identified to be components of the protein-bound polysaccharide analyzed by automated amino acid analyzer. Antioxidant activity was evaluated at different concentrations ranged from 0.25 to 3 mg/ml using three different in vitro antioxidant activity assays. The obtained results clearly indicated that the MeOH extract and PAPs showed a strong antioxidant activity in terms of FRAP, DPPH• free radical scavenging and reducing power assay. Thus, Plumeria alba could be a new source of natural antioxidants for functional foods or medicine due to its higher antioxidant potential, flavonoids and phenolics contents.

Deproteinization and structural characterization of bioactive exopolysaccharides from Ganoderma sinense mycelium

Deproteinization was comparatively employed for the purification of crude exopolysaccharide (CEP) of Ganoderma sinense mycelium. Results showed that ammonium sulfate (70% saturation) salting out could remove 95% proteins of CEP, whereas 20% trichloroacetic acid precipitation was best with a polysaccharide loss less than 20%. Protein-bound polysaccharides could be removed by both methods, and polysaccharides with high molecular weight dominated in the lost polysaccharides. Through anion-exchange chromatography, a highly branched α-(1→6)-D-mannan and a protein-bound heteropolysaccharide were prepared from CEP. Both fractions could inhibit the in vitro proliferation of tumor cells BEL-7402, and induce TNF-α secretion of murine splenocytes.

Characterisation of bioactive protein-bound polysaccharides from Amanita ponderosa cultures

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A study on immunomodulatory mechanism of Polysaccharopeptide mediated by TLR4 signaling pathway.

BackgroundPolysaccharopeptide (PSP), isolated from Coriolus versicolor COV-1 strain, is a protein-bound polysaccharide widely used as immunoadjuvant for cancer immunotherapy. Although the immunomodulatory activity of PSP has been well established, the precise molecule mechanisms of its biological activity have yet to be fully elucidated.MethodsIn the present study, we first investigated the immunomodulatory activity of PSP in peritoneal macrophages from C57BL/10J (TLR4+/+) and C57BL/10ScCr (TLR4-/-) mice carrying a defective toll-like receptor-4 (TLR4) gene and then evaluated PSP for its effect on tumor inhibition rates and the immune organ index in above two different strains of mice. In addition, PSP were also evaluated for its activation of TLR4, TLR4-downstream molecules (TRAF6, NF-κB and AP-1) in spleens of tumor-bearing C57BL/10J (TLR4+/+) and C57BL/10ScCr (TLR4-/-) mice.ResultsThe results showed that PSP had adjuvant activities in stimulating expressions of cytokines as well as TLR4, TRAF6, phosphorylation of NF-κB p65 transcription factors and phosphorylation of c-Jun (a component of the transcription factor AP-1) in peritoneal macrophages from C57BL/10J (TLR4+/+) mice but not from C57BL/10ScCr (TLR4-/-) mice. In vivo PSP as well as Adriamycin (ADM) decreased the mean weights of tumors compared with normal saline and PSP increased thymus index and spleen index relative to ADM in tumor-bearing C57BL/10J (TLR4+/+) mice but not in C57BL/10ScCr (TLR4-/-) mice.ConclusionsWe demonstrated that PSP activates peritoneal macrophages in vitro via TLR4 signaling pathway and PSP functions its immunoregulatory effect in vivo also via TLR4 signaling pathway. These data strongly suggest TLR4 signaling pathway is involved in PSP-mediated immunomodulatory activities.

Clinical efficacy of protein-bound polysaccharide K in patients with gastric cancer undergoing chemotherapy with an oral fluoropyrimidine (S-1)

BackgroundThe aim of the present study was to evaluate the clinical significance of protein-bound polysaccharide K (PSK) in patients with primary gastric cancer who were being treated with an oral fluoropyrimidine (S-1). MethodsClinical reports of 190 gastric cancer patients treated with S-1 chemotherapy, with or without PSK, at Kochi Medical School between 2007 and 2012 were investigated retrospectively to analyze survival. The neutrophil:lymphocyte ratio (NLR) was also evaluated as indicator of the immunoenhancing effect of PSK. ResultsOverall survival was significantly longer in patients treated with S-1 + PSK than in those given S-1 alone (hazard ratio for death, 0.608; 95% confidence interval 0.375–0.985; P = 0.041). Furthermore, there was a tendency for changes in the NLR during chemotherapy to be lower in the S-1 + PSK group than in the S-1 group, but the difference did not reach statistical significance (P = 0.054). When patients were divided into groups based on preoperative NLR (i.e. <2.5 and ≥2.5), the mean (±SEM) NLR 1 month after the beginning of chemotherapy in the NLR ≥2.5 subgroup was significantly lower in patients treated with S-1 + PSK rather than S-1 alone (1.7 ± 0.7 vs. 3.3 ± 4.1, respectively; P = 0.043). ConclusionsImmunochemotherapy using PSK improves the survival of patients with advanced gastric cancer. The NLR may be a useful biomarker for evaluating prognosis in these patients.

Rapeseed polysaccharides as prebiotics on growth and acidifying activity of probiotics in vitro

In vitro digestibility, prebiotic activity and chemical composition of polysaccharides from rapeseed were deliberately studied in this paper. After preliminary treatments, two fractions of polysaccharides (RP1 and RP2) were obtained after purification by DEAE-cellulose and Sephadex G-100. Their primary structural feature and molecule weights were characterized. Furthermore, their digestibility was also evaluated by artificial gastric juice and α-amylase. Finally, their proliferative effect on bifidobacteria and lactobacilli and acid production of the resulting probiotics in vitro were investigated. The results showed that RP1 and RP2 were homogeneously protein-bound polysaccharides with molecular weights of 28.51 and 6.55kDa, respectively. They were resistant to hydrolysis by artificial gastric juice and α-amylase. Moreover, they could also significantly stimulate the tested probiotics to proliferate and produce organic acids. These findings clearly suggest the polysaccharides from rapeseed are potential to be exploited as novel prebiotics.

Multicenter trial for assessing cytokine promoter gene polymorphism as a predictive parameter of PSK responder for curatively resected stage II or III colorectal cancer.

619 Background: Polysaccharide-K (PSK), a protein-bound polysaccharide extracted from the mycelia of Coriolus versicolor, is an immunomodulator widely used in colorectal cancer in Japan. PSK has immunological actions including enhancement or inhibition of cytokine production. It has been reported that levels of cytokine production are influenced by polymorphisms in the promoters of cytokine genes. We hypothesized that cytokine promoter gene polymorphisms may be responsible for genetic susceptibilities to immunological effect of PSK. Methods: This is a multicenter prospective trial. One hundred and ten patients with stage II or III colorectal cancer were enrolled. All patients received adjuvant immnochemotherapy after curative resection using UFT (stage II) or UFT/LV (stage III) combined with PSK (3.0 g/day, p.o.) for 1 year. Post-operative survey of recurrence was followed with CT scan at 6-month intervals during the first 2 years after surgery and at 1-year intervals thereafter until 5 year after surgery. DNA was extracted from peripheral blood cells in all patients. The following polymorphisms of the patients were genotyped: TNF-α-1031T/C, IL-1ß-511C/T, IL-6-634C/G, IL-10-819T/C. Results: Twenty (6 in stage II, 14 in stage III) out of 110 patients showed recurrence after more than 5 years survey. Fifteen out of 74 patients with TNF-α-1031 TT genotype and 5 out of 36 with TNF-α-1031 CC or CT showed disease recurrence. Five out of 39 patients with IL-1ß-511 CC and 15 out of 71 with IL-1ß-511 CT or TT showed disease recurrence. Eleven out of 61 patients with IL-6-634 CC and 9 out of 49 with IL-6-634 CG or GG showed disease recurrence. No association between genotype frequency and disease recurrence was observed for TNF-α, IL-1ß, IL-6. Fifteen out of 61 patients with IL-10-819 CC or CT genotype showed disease recurrence, whereas only 5 out of 49 with IL-10-819 TT showed tumor recurrence (p=0.052). Especially, significant association with disease recurrence was found in stage III patients (12/30 vs 2/17, p=0.042). Conclusions: It is suggested that colorectal cancer patients with IL-10-819 TT genotype could be PSK responder after curative resection.

Protein-bound polysaccharide K suppresses tumor fibrosis in gastric cancer by inhibiting the TGF-β signaling pathway.

Peritoneal carcinomatosis (PC) is the most frequent metastatic pattern of gastric cancer and its prognosis is extremely poor. PC is characterized by rich fibrosis and the development of obstructive disorders such as ileus, jaundice and hydronephrosis. Epithelial-mesenchymal transition (EMT) is one of the major causes of tissue fibrosis and transforming growth factor β (TGF-β) has a pivotal function in the progression of EMT. Protein-bound polysaccharide K (PSK) is a biological response modifier that can modulate the TGF-β/Smad signaling pathway in vitro. In the present study, we established a fibrotic tumor model using human peritoneal mesothelial cells (HPMCs) and a human gastric cancer cell line to evaluate whether PSK attenuates tumor fibrosis. HPMCs exposed to PSK did not undergo the morphological change from a cobblestone-like pattern to a spindle-shape pattern normally induced by treatment with TGF-β. Immunofluorescence further demonstrated that PSK suppressed TGF-β-induced overexpression of α-SMA in the HPMCs. We further showed that HPMCs contributed to the proliferation of tumor fibrosis by using a mouse xenograft model. Additionally, PSK treatment of these mice significantly reduced the area of observable tumor fibrosis. These results suggest that seeded cancer cells transformed HPMCs into myofibroblast-like cells through their release of TGF-β in the microenvironment, facilitating the development of fibrous tumors in organs covered with HPMCs. Therefore, our study indicates that PSK has potential utility as an anti-fibrotic agent in the treatment of gastric cancer patients with PC.

팽이버섯 자실체, 팽이버섯 균사체 및 동충하초 균사체의 유용성분 분석

일반성분 중 조회분 함량은 버섯이 균사체 보다 높은 조회분 함량을 보였다. 조단백질은 모든 시험구에서 19% 내외로 나타났고, 조지방 함량은 동충하초 균사체에서 가장 높았으며, 조섬유는 팽이버섯에서 가장 높게 나타났다. 가용성물질소물은 팽이버섯균사체와 동충하초균사체에서 60% 이상의 함량을 나타내었다. 각 시료별 총단백질과 총당의 구성 비율은 팽이버섯과 팽이버섯 균사체의 단백다당류의 구성이 다름을 확인하였다. 시료별 핵산관련물질 함량은 팽이버섯에서 5'-GMP, 5'-XMP, 5'-IMP순으로 높게 나타났고, 팽이버섯 균사체는 5'-XMP, 5'-GMP, 5'-IMP, 동충하초 균사체는 5'-XMP, 5'-IMP, 5'-GMP순으로 높게 나타났다. 모든시료에서 각각 17종의 아미노산이 검출되었다. 각 시료의 총 아미노산 함량을 살펴보면 팽이버섯 18,980 mg%, 팽이버섯 균사체 17,604 mg%, 동충하초 균사체 18,429 mg%로 나타났고 모든 시료에서 glutamic acid 함량이 가장 높았다. 유리아미노산 함량도 구성아미노산과 유사하게 팽이버섯, 팽이버섯 균사체, 동충하초 균사체 순으로 높게 나타났다. 본 실험결과 팽이버섯, 팽이버섯 균사체 및 동충하초 균사체는 핵산관련 물질과 아미노산 함량이 높게 나타나, 아미노산 함유 식품 개발에 적합한 자원으로 활용도가 기대된다. Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium were analyzed for their proximate composition, protein-bound polysaccharide, nucleic acid and amino acids. The content of ash and crude fiber in F. velutipes fruit body were higher than F. mycelium and C. militaris mycelium. C. militaris mycelium showed the highest crude fat content while F. velutipes fruit body had lowest. Nitrogen free extract content of the samples varied from 56.8% in F. velutipes fruit body to 61.9% in F. velutipes mycelium. The compositions of total protein and total free sugars of protein-bound polysaccharide were found to be significant differences for all samples. Nucleic acid related compounds were identified the 5'-GMP, 5'-XMP, 5'-IMP in all samples. The content of total nucleic acids were high in the orders of F. velutipes myclial (286.71 mg%), F. velutipes fruit body(187.36 mg%) and C. militaris mycelial(76.85 mg%). The highest content of 5'-GMP was found in F. velutipes fruit body. The most nucleic acid of F. velutipes mycelial and C. militaris mycelial were the 5'-XMP. As for the analysis of total amino acids, seventeen amino acids were identified by HPLC and the major amino acid was glutamic acid in all samples. The content of total amino acids were high in the orders of F. velutipes fruit body(19,919 mg%), F. velutipes mycelium(19,018 mg%) abd C. militaris mycelium(18,965 mg%). We determined the developing new food product such as amino acid drink and amino acid containing food using extracts of Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium.

Hepatoprotective bioactivity of the glycoprotein, antrodan, isolated from Antrodia cinnamomea mycelia.

Antrodan, a protein-bound polysaccharide isolated from Antrodia cinnamomea mycelia, was demonstrated to exhibit significant anti-inflammatory bioactivity in vitro. However, its role in hepatic injury in vivo still remains unclear. We hypothesized that antrodan may have beneficial hepatoprotective effects. To verify this, a lipopolysaccharide (LPS)-Sprague-Dawley rat model was used. Antrodan protected against liver damage by suppressing LPS-stimulated serum glutamine-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), interleukin (IL)-6, hepatic thiobarbituric acid reactive substances (TBARS), nitric oxide (NO), inducible NO synthase (iNOS) and nuclear factor (NF)-κB, and by effectively alleviating the downregulated hepatic superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH-Px). Hematoxylin-eosin staining revealed that antrodan at a dosage of 40 mg/kg was able to alleviate LPS-induced liver damage to a normal status. In addition, we identified the partial main architectural backbone of antrodan to have a 1→3 linear β-glycosidic backbone of mannan linked by β-1→3 glucosidic branches. Conclusively, antrodan can potentially ameliorate liver damage in vivo by suppressing oxidative stress induced by LPS.

Immunomodulatory activities of polysaccharides isolated from Taxillus chinensis and Uncaria rhyncophylla

Taxillus chinensis and Uncaria rhyncophylla are the herbs used in traditional Chinese anticancer formulations. During the past decade, research on plant polysaccharides has gained importance due to their therapeutic value and minimum side effects. In this study, hot water extraction method was employed to isolate polysaccharides from the stems of T. chinensis and stems with hooks of U. rhyncophylla. Size-exclusion chromatography was then used for further fractionation. Separated fractions from T. chinensis were designated as TCP-1, TCP-2 and TCP-3 and those from U. rhyncophylla were termed UC-1 and UC-2. Their sugar compositions were estimated using gas chromatography that revealed the presence fructose, glucose, xylose, arbinose, and rhamnose. Amino acid analysis of these fractions has indicated that they are protein-bound polysaccharides. The antioxidant activities were investigated using DPPH and yeast assays. The ability of these polysaccharide fractions to stimulate mouse macrophages was measured using Griess reagent and ELISA test. The results revealed that some of the isolated fractions (TCP-2, TCP-3, UC-1 and UC-2) displayed significant antioxidant activities and were also found to be effective immunomodulators in a concentration-dependent manner. Outcomes of this research strongly indicate that U. rhyncophylla and T. chinensis have therapeutic potential to be used for the treatment of cancer.

Structure characterization and immunocompetence of a glucan from the fruiting bodies of Cantharellus cibarius

A protein-bound polysaccharide fraction (JBP-1) was obtained from the fruiting bodies of Cantharellus cibarius. Its chemical composition was studied by the cooperative usage of multiple chemical and spectral methods and characterized to be a fraction with a molecular weight of 4.8 × 105 Da and only composed of glucose. Methylation analysis revealed that the sugar residues in JBP-1 are existing as t-, 1,6-, and 1,3,6-linked Glcp sugar residues. The immunocompetence of the fraction was evaluated with the proliferation assay of mouse splenocytes, and the result revealed that JBP-1 could significantly stimulate the proliferation of mouse splenocytes in a dose-dependent manner, with p < 0.001 at the concentration of 100 μg/ml and 30 μg/ml, p < 0.05 at 10 μg/ml. These results give us a primary scientific evidence to further explore the pharmaceutical function of this mushroom.

Augmentation of Antitumor Immunity by Fusions of Ethanol-Treated Tumor Cells and Dendritic Cells Stimulated via Dual TLRs through TGF-β1 Blockade and IL-12p70 Production

The therapeutic efficacy of fusion cell (FC)-based cancer vaccine generated with whole tumor cells and dendritic cells (DCs) requires the improved immunogenicity of both cells. Treatment of whole tumor cells with ethanol resulted in blockade of immune-suppressive soluble factors such as transforming growth factor (TGF)-β1, vascular endothelial growth factor, and IL-10 without decreased expression of major histocompatibility complex (MHC) class I and the MUC1 tumor-associated antigen. Moreover, the ethanol-treated tumor cells expressed “eat-me” signals such as calreticulin (CRT) on the cell surface and released immunostimulatory factors such as heat shock protein (HSP)90α and high-mobility group box 1 (HMGB1). A dual stimulation of protein-bound polysaccharides isolated from Coriolus versicolor (TLR2 agonist) and penicillin-inactivated Streptococcus pyogenes (TLR4 agonist) led human monocyte-derived DCs to produce HSP90α and multiple cytokines such as IL-12p70 and IL-10. Interestingly, incorporating ethanol-treated tumor cells and TLRs-stimulated DCs during the fusion process promoted fusion efficiency and up-regulated MHC class II molecules on a per fusion basis. Moreover, fusions of ethanol-treated tumor cells and dual TLRs-stimulated DCs (E-tumor/FCs) inhibited the production of multiple immune-suppressive soluble factors including TGF-β1 and up-regulated the production of IL-12p70 and HSP90α. Most importantly, E-tumor/FCs activated T cells capable of producing high levels of IFN-γ, resulting in augmented MUC1-specific CTL induction. Collectively, our results illustrate the synergy between ethanol-treated whole tumor cells and dual TLRs-stimulated DCs in inducing augmented CTL responses in vitro by FC preparations. The alternative system is simple and may provide a platform for adoptive immunotherapy.

Multicenter trial for assessing tolerability of combination therapy with cisplatin, irinotecan, and PSK in extensive-stage small-cell lung cancer: RNCLC-01 study.

3072 Background: Polysaccharide-K (PSK, Krestin) is a protein-bound polysaccharide, extracted from cultured mycelium of Coriplus versicolor, and is associated with immunostimulatory activity. Although combination use of PSK with chemotherapy has been shown to prolong the response period in patients with small-cell lung cancer (SCLC), the efficacy of PSK in combination chemotherapy including cisplatin has been less examined. We examined safety and efficacy of combination therapy with cisplatin, irinotecan, and PSK in extensive-stage (ED) SCLC. Methods: Eligible pts included: histologically confirmed ED-SCLC without prior chemotherapy, age under 75 years, ECOG performance status ≤ 1, with evaluable disease and adequate organ function. Treatment: irinotecan 60 mg/m2 on days 1, 8, and 15 and cisplatin 60 mg/m2 on day 1 every 4 weeks for 4 to 6 courses, and oral PSK 3,000 mg/body daily. The treatment with PSK was continued after the end of cisplatin and irinotecan administrations, and was combined as a part of second-line therapy after exacerbations. Results: Between January 2008 and July 2010, 17 patients were enrolled, and the efficacy and prognosis were evaluated in 15 of 17 patients. Response rate was 66.6%, one-year survival rate was 66.6%, and two-year survival rate was 33.2%. The major toxicities were diarrhea and myelosuppression, and the common toxicity of PSK-containing combination chemotherapy was not observed. Conclusions: The combination chemotherapy with cisplatin, irinotecan, and PSK was effective and well tolerated in ED-SCLC. Clinical trial information: NCT00546130.