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Objective:To investigate the antigen-sparing effects of crude polysaccharides from Cistanche deserticola Y. C.Ma (CPCD) for influenza virus vaccine (IVV). Methods:ICR mice were immunized subcutaneously with CPCD combined with different doses of IVV (0.01 μg and 0.1 μg). Hemagglutinin inhibition (HI) assay was used to detect HI titers in serum samples. Indirect ELISA was performed to detect the levels of specific IgG antibodies and their subtypes in serum samples. The proliferation of splenic lymphocytes was detected by MTT assay. The percentages of CD4 + , CD8 + and CD44 + T cells and the levels of IFN-γ in splenic cells isolated from the vaccinated mice were analyzed by flow cytometry. Results:CPCD significantly increased HI titers (234.67±47.70 vs 149.33±47.70, P<0.05), promoted the production of IgG ( A450 value: 1.16±0.63 vs 0.30±0.21, P<0.05) and IgG1 ( A450 value: 1.09±0.60 vs 0.26±0.21, P<0.05) and enhanced splenic lymphocyte proliferation ( P<0.05). CPCD also significantly up-regulated the expression of CD4 + [(41.97±4.58)% vs (25.43±1.48)%, P<0.05], CD8 + [(12.67±0.33)% vs (9.02±1.07)%, P<0.05], CD4 + CD44 + [(11.77±0.69)% vs (8.64±0.71)%, P<0.05] and CD8 + CD44 + [(6.70±0.67)% vs (4.66±0.39)%, P<0.05] T cell subsets as well as the secretion of IFN-γ in CD4 + [(1.36±0.07)% vs (0.87±0.06)%, P<0.05] and CD8 + [(2.09±0.20)% vs (1.42±0.08)%, P<0.05] T cells. In addition, there was no significant difference between CPCD combined with low-dose IVV group and high-dose IVV alone group ( P>0.05), implying a 10-fold antigen sparing. Conclusions:CPCD, as an adjuvant for influenza virus vaccine, could enhance humoral and cellular immune responses and reduce antigen dose, which might be a potential adjuvant for seasonal or pandemic influenza vaccines.
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Objective:To investigate the optimal dose of Th1/Th2 immuno-enhancement effects of cultivated Artemisia rupestris L. crude polysaccharides (CARCP) on foot-and-mouth disease vaccine (FMDV) via the intramuscular route. Methods:ICR mice were intramuscularly immunized twice with different concentrations of CARCP mixed with FMDV at 2-week intervals. FMDV-specific antibodies, isotypes, and IgE in serum were detected by ELISA. Splenocyte proliferation was detected by MTT. T lymphocyte subsets and cytokines in the spleen were detected by flow cytometry. Clinical signs and local reactions at the injection site were monitored daily, and the body weight of mice was weighed after immunization.Results:The medium dose of CARCP could significantly improve FMDV-specific IgG, IgG 1, and IgG 2a antibody levels and the IgG 2a/IgG 1 ratio ( P<0.05) and lead to significant splenocyte proliferative responses ( P<0.01). The medium dose of CARCP could also significantly increase the level of CD3 +CD4 + and CD3 +CD8 + T cells as well as CD4 +/CD8 + ratio ( P<0.05), elicited the higher levels of IFN-γ in CD4 + T cells and CD8 + T cells ( P<0.05). No local adverse reactions at the injection site were observed after immunization. There was no significant difference in body weight or growth between each group( P>0.05). CARCP did not significantly induce an IgE response ( P>0.05). Conclusions:CARCP as an FMDV adjuvant promotes Th1/Th2 immune responses, especially in favor of the Th1 response, and has a certain safety profile. The best immune enhancement is achieved with CARCP at medium doses.
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BACKGROUND: Cigarette smoke contains a large number of types of oxygen free radicals and cytotoxic components. Passive smoking will impair respiratory and cardiovascular system functions, and result in oxidative damage of skeletal muscle and decreased exercise ability. OBJECTIVE; To investigate the effect of phellinus igniarius crude polysaccharides on the exercise capacity and free radical metabolism of skeletal muscle in mice suffering passive smoking, so as to provide ideas for the prevention and treatment of peroxidative damage of skeletal muscle and depression of exercise capacity in rats suffering passive smoking. METHODS: Twenty-one male Kunming mice were randomly assigned to three groups: Gavage with phellinus igniarius crude polysaccharides and suffering passive smoking (phellinus group), gavage with distilled water and suffering passive smoking (control group), and only gavage with distilled water (blank group). After 4 consecutive weeks, the mice were forced to take an exhausted swimming, and sacrificed subsequently. Exhausted swimming time was recorded. The bilateral gastrocnemius muscle tissues were obtained, in which the vitality of superoxide dismutase, catalase, glutathion reductase and Ca2+-Mg2+-ATP and Na+-K+-ATP activity, and the concentration of malonaldehyde were measured. RESULTS AND CONCLUSION: (1) The swimming time of mice in the control group was shorter than that in the blank group (P 0.6, P 0.6, P < 0.05), and negatively correlated with the concentration of malonaldehyde (r < -0.6, P < 0.05). (5) In summary, phellinus igniarius crude polysaccharides can improve the antioxidative enzyme activity of skeletal muscle, inhibit lipid peroxidation reaction, and thus increase exercise ability of mice suffering passive smoking. The study was approved by the Ethical Committee of Jiangxi Normal University, approval No. 201703.
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Objective To investigate the enhancement effect of Xinjiang wild Artemisia rupestris L. crude polysaccharides (WARCP) as an adjuvant on ovalbumin (OVA) vaccine in mice immunized intramuscularly. Methods ICR mice were randomly divided into 6 groups (5 per group), including 9 g/L NaCl group (blank control), OVA group (10 μg OVA), low dose WARCP/OVA group (OVA+50 μg WARCP), medium dose WARCP/OVA group (OVA+200 μg WARCP), high dose WARCP/OVA group (OVA+400 μg WARCP), and aluminum adjuvant (Alum)/OVA group (positive control group, OVA +100 μg Alum). ICR mice were immunized intramuscularly and weighted. The OVA-specific antibodies and subtypes in serum were detected by enzyme linked immunosorbent assay (ELISA). T cells subsets from spleen and lymph nodes were detected by flow cytometry. Results The medium-dose WARCP/OVA group enhanced IgG and IgG1 levels and increased early antibody levels (all P<0.05). The medium-dose WARCP/OVA group and the high-dose WARCP/OVA group significantly enhanced IgG2a levels (all P<0.05), but the difference was not statistically significant comparing with Alum/OVA group (P>0.05). The low-dose WARCP/OVA group enhanced the percentage of CD4+ T cells in spleen and CD4 + T, CD8+ T, CD4 +CD44 + T cells in lymph nodes (all P<0.05). The medium dose WARCP/OVA group and the high dose WARCP/OVA group enhanced the CD4 + T, CD8 + T, CD4 +CD44 + T, CD8 +CD44+ T cells in spleen and CD8+CD44+ T cell in lymph nodes (all P<0.05). Conclusions Plant-derived WARCP as an OVA protein vaccine adjuvant can enhance cellular immunity and humoral immunity, and it is safe and reliable. The results in this study provide a theoretical basis for the popularization and application of WARCP.
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Objective@#To investigate the enhancement effect of Xinjiang wild Artemisia rupestris L. crude polysaccharides (WARCP) as an adjuvant on ovalbumin (OVA) vaccine in mice immunized intramuscularly.@*Methods@#ICR mice were randomly divided into 6 groups (5 per group), including 9 g/L NaCl group (blank control), OVA group (10 μg OVA), low dose WARCP/OVA group (OVA+50 μg WARCP), medium dose WARCP/OVA group (OVA+200 μg WARCP), high dose WARCP/OVA group (OVA+400 μg WARCP), and aluminum adjuvant (Alum)/OVA group (positive control group, OVA+100 μg Alum). ICR mice were immunized intramuscularly and weighted. The OVA-specific antibodies and subtypes in serum were detected by enzyme linked immunosorbent assay (ELISA). T cells subsets from spleen and lymph nodes were detected by flow cytometry.@*Results@#The medium-dose WARCP/OVA group enhanced IgG and IgG1 levels and increased early antibody levels (all P<0.05). The medium-dose WARCP/OVA group and the high-dose WARCP/OVA group significantly enhanced IgG2a levels (all P<0.05), but the difference was not statistically significant comparing with Alum/OVA group (P>0.05). The low-dose WARCP/OVA group enhanced the percentage of CD4+ T cells in spleen and CD4+ T, CD8+ T, CD4+CD44+ T cells in lymph nodes (all P<0.05). The medium dose WARCP/OVA group and the high dose WARCP/OVA group enhanced the CD4+ T, CD8+ T, CD4+CD44+ T, CD8+CD44+ T cells in spleen and CD8+CD44+ T cell in lymph nodes (all P<0.05).@*Conclusions@#Plant-derived WARCP as an OVA protein vaccine adjuvant can enhance cellular immunity and humoral immunity, and it is safe and reliable. The results in this study provide a theoretical basis for the popularization and application of WARCP.
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Objective To investigate the effects of Glycyrrhiza uralensis Fisch L. crude polysac-charides (GUCP) as an adjuvant on the immunity of mice and the immune responses induced by human pap-illoma virus ( HPV)-DNA vaccine. Methods ICR mice were injected with different concentrations of GUCP by different ways to detect the influences of GUCP on body weight, organ indexes and the numbers of immune cells in spleen. C57BL/ 6 mice were co-immunized with HPV-DNA vaccine and GUCP to detect the adjuvant efficacy on antigen-specific cellular and humoral immune responses. Results GUCP in all injected groups had no side effect on mouse body weight and liver, heart, lung and kidney indexes, but intraperitone-al injection of GUCP significantly increased spleen and thymus indexes and the numbers of B cells, CD4+ T cells, macrophages and dendritic cells in spleen. Subcutaneous injection of GUCP significantly increased the numbers of B cells and macrophages in spleen and intragastric administration significantly increased the num-bers of CD4+ and CD8+ T cells in spleen. Furthermore, GUCP as an adjuvant enhanced the antigen-specific CD4+ and CD8+ T cell responses and the levels of IgG, IgG1 and IgG2a induced by HPV-DNA vaccine at a certain degree. Conclusion GUCP enhanced the immunity of mice and the antigen-specific cellular and hu-moral immune responses induced by HPV-DNA vaccine. These results suggested that GUCP might be used as an adjuvant for DNA vaccine.
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An Affi-Prep Polymyxin column was combined with a Phenyl Sepharose column and a Sephacryl S-300 column, respectively, to remove the lipopolysaccharides(LPS) in the anti-complementary crude polysaccharides of Houttuynia Herba. The contents of LPS in the polysaccharides were determined by chromogenic tachypleus amebocyte lysate(TAL)method during the procedure of purifying. The anti-complementary activities of the polysaccharides were also compared before and after the removal of LPS. Less remanent LPS was detected after purified using Penyl Sepharose combined with polymyxin column, with the clearance rate of 42.85%. All the columns had no effect on the anti-complementary activity of the polysaccharides. Penyl Sepharose combined with polymyxin column would be sound for LPS removal of the anti-complementary polysaccharides without reducing their bioactivity.
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Objective To investigate the immunopotentiating effects of cultivated Cistanche deser-ticola (C.deserticola) crude polysaccharides (CCDCP) as an adjuvant on the model antigen ovalbumin (OVA). Methods Low,medium and high doses of CCDCP in combination with OVA were intramuscularly injected twice into ICR mice at an interval of two weeks,respectively. Aluminum adjuvant was used to set up positive control group. Levels of IgG,IgG1 and IgG2a antibodies were detected by ELISA. Splenocyte prolif-eration was detected by MTT assay. Growth conditions of the immunized mice were observed. Results IgG level was significantly increased in the high dose group 7 days after the first immunization(P<0.05),espe-cially on the 21st and 28th days (P<0.01) as compared with that of the aluminum adjuvant group. High dose of CCDCP in combination with OVA significantly up-regulated the levels of IgG1 and IgG2a in mice as compared with immunization with OVA alone (P<0.05). Moreover, IgG2a level in mice immunized with high dose of CCDCP and OVA was higher than that of the aluminum adjuvant group(P<0.05). Splenocyte proliferation was significantly enhanced in the medium and low dose groups in comparison with that of the OVA group (P<0.05) and the aluminum adjuvant group (P<0.01). No significant difference in mouse body weight was observed in different groups(P>0.05). Conclusion CCDCP as an adjuvant of OVA pro-tein vaccine can enhance Th1 and Th2 immune responses,especially the early antibody production and Th1 immune response. These results will provide some information for further studies of CCDCP as a vaccine ad-juvant.
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Objective To investigate the efficacy of using crude polysaccharides extracted from cultivated Artemisia rupestris L. ( CARCP) in Xinjiang as an immunologic adjuvant for ovalbumin ( OVA) . Methods The mice were subcutaneously immunized twice with OVA vaccine formulated with CARCP. ELISA assay was performed to measure the levels of IgG, IgG1 and IgG2a antibodies. Flow cytometry analy-sis was performed to detect the percentages of different T lymphocyte subsets ( CD3+CD4+ and CD3+CD8+T cells, CD4+CD44+ and CD8+CD44+T cells) in splenocytes as well as the expression of intracellular cyto-kines ( CD4+IFN-γ, CD8+IFN-γ) and CD4+CD25+Foxp3+Treg cells. Results The levels of IgG, IgG1 and IgG2a antibodies in serum, the percentages of CD3+CD4+, CD3+CD8+, CD4+CD44+ and CD8+CD44+T cells as well as the ratio of CD4+/CD8+T lymphocytes increased significantly in mice immunized with OVA in combination with CARCP (P<0. 05). Moreover, CARCP enhanced the expression of CD4+IFN-γand CD8+IFN-γ( P<0. 05 ) , but inhibited the expression of CD 4+CD 2 5+Foxp 3+Treg cells ( P<0. 05 ) . Conclusion As an adjuvant for OVA, CARCP enhanced the humoral and cellular immune responses, especially the T cell immune responses.
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OBJECTIVE:To optimize the extraction technology of Compound epimedium chewable tablets. METHODS:Using the extraction rate of icariin,the content of crude polysaccharide and yield of extractum as index,the effects of water amount,ex-traction time and extraction times on extraction effect were investigated by orthogonal test. The extraction technology was optimized and validation test was conducted. RESULTS:The optimized extraction technology was as follows as 6-fold water,extracting 3 times,2 h each time. The validation test showed that the average extraction rate of icariin was(83.80±0.02)%,the average con-tent of crude polysaccharide was (29.28 ± 0.55) mg/g,and the average yield of extractum was (28.47 ± 0.29)%(RSD≤0.55%, n=3). CONCLUSIONS:The optimized extraction technology is stable,and can be used for the extraction of Compound epimedi-um chewable tablets.
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Objective To study the optimum decoloration technological conditions of crude polysaccharides from Cordyceps militaris Link. (PCM) by macroporous resin. Methods With the decolorization rate and retention rate of PCM as indexes, static adsorption factor experiment was used for the optimization of seven kinds of macroporous resin. And then the optimum conditions of decolorization of PCM were investigated by the static/dynamaic adsorption single-factor experiments. Results D941 macroporous resin is the best. The optimum decoloration technological conditions were as follows:initial concentration of the sample was 50 mg/mL, pH value was 5.5, operating temperature was 45℃, the flow rate of dynamaic adsorption was 2.5 BV/h, and the sample solution volume was 5 bed volume per hour. Under the optimal conditions, decolorization rate and retention rate of PCM were ( 93.9 ± 3.4) % and (91.7 ± 2.2) %, respectively. Conclusion The resin D941 is suitable for the decoloration of PCM. The optimized method is feasible and reliable, having the prospects for popularizing.
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Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. beta-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of 10~2,000 microg/mL. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of 50 microg/mL, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at 200 microg/mL. Maximum production of nitric oxide (14.3 microM) was recorded in the Fr. NaCl fraction at 200 microg/mL. Production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-alpha, IL-1beta, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.
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Animals , Mice , Agaricales , Alkaline Phosphatase , B-Lymphocytes , Cell Line , Fruit , Immunomodulation , Injections, Intraperitoneal , Interleukin-1beta , Interleukin-6 , Korea , Life Support Care , Methanol , Nitric Oxide , Pleurotus , Polysaccharides , Sarcoma , Sarcoma 180 , Spleen , Tumor Necrosis Factor-alpha , WaterABSTRACT
This study was initiated in order to investigate the anticancer and immunomodulating activities of crude polysaccharides extracted in methanol, neutral saline, and hot water (hereinafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of Panellus serotinus. Content of beta-glucan and protein in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. serotinus ranged from 22.92~28.52 g/100 g and 3.24~3.68 g/100 g, respectively. In vitro cytotoxicity tests, none of the various fractions of crude polysaccharides were cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at the tested concentration. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 23.53~44.71% in mice previously inoculated with sarcoma 180. Treatment with Fr. HW resulted in an increase in the numbers of spleen cells by 1.3 fold at the concentration of 50 microg/mL compared with control. Treatment with Fr. NaCl resulted in improvement of the immuno-potentiating activity of B lymphocytes by increasing the alkaline phosphatase activity by 1.4 fold, compared with control, at the concentration of 200 microg/mL. Among the three fractions, maximum nitric oxide (13.48 microM) was recorded at 500 microg/mL in Fr. HW. Production of tumor necrosis factor alpha, interleukin-1beta, and interleukin-6 was significantly higher, compared to the positive control, concanavalin A, at the tested concentration. Therefore, treatment with crude polysaccharides extracted from the fruiting body of P. serotinus could result in improvement of antitumor activity.
Subject(s)
Animals , Mice , Alkaline Phosphatase , B-Lymphocytes , Cell Line , Concanavalin A , Fruit , Injections, Intraperitoneal , Interleukin-1beta , Interleukin-6 , Life Support Care , Methanol , Nitric Oxide , Polysaccharides , Sarcoma 180 , Spleen , Tumor Necrosis Factor-alpha , WaterABSTRACT
Objective To investigate the glycosidic linkage and immunomodulating activities of crude polysaccharides from the seeds of Zea mays. Methods Extraction, isolation and purification of polysaccharides were carried out with boiling-water extraction plus resolving deposit repeatedly. The structure was elucidated on basis of physicochemical properties and spectral data, and the biological activities were evaluated by means of Immunopharmacological examination. Results The structure of polysaccharides from the seeds of Z. mays exhibited identical structure with rice bran polysaccharides, i.e., a kind of glucan withα-1,4 andα-1,6 glucosidic bonds as the main frame. Conclusion Polysaccharides was obtained from the seeds of Z. mays for the first time, and it showed significant immunomodulating activity in mice.