Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts / 한국영양학회지

PURPOSE: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models.METHODS: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit.RESULTS: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4⁺ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group.CONCLUSION: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

Immune stimulatory activity of BRP-4, an acidic polysaccharide from an edible plant, Basella rubra L

OBJECTIVE@#To evaluated the immunomodulatory effect of BRP-4, an acidic polysaccharide from Basella rubra (B. rubra) L on the macrophage activity.@*METHODS@#Phagocytic activity was determined by the ingestion of Latex Beads-Rabbit IgG-FITC using the fluorescent microscopy and flow cytometry analysis and nitric oxide production was measured using Griess reaction assay.@*RESULTS@#An enhanced production of NO was observed at 10 and 100 μg/mL of BRP-4. The phagocytic activity of macrophage was enhanced in BRP-4 treated RAW264.7 cells. BRP-4 combined with concanavalin A (Con A) provided obvious promotion and strengthening of the proliferation of the splenocytes.@*CONCLUSIONS@#BRP-4, polysaccharide isolated from B. rubra, is suggested to activate macrophage function and stimulate splenocyte proliferation. The strong immunomodulatory activity of BRP-4 confirmed its good potential as an immunotherapeutic adjuvant.

Dynamic observation on splenocyte in mice immunized with recombinant Bi f idobacterium bi f idum (pGEX-S j26GST) of Schistosoma j aponicum / 中国人兽共患病学报

To observe the dynamic changes of splenocyte proliferation ,subsets and apoptosis in mice immunized with re-combinantBifidobacteriumbifidum(pGEX-Sj26GST)ofSchistosomajaponicum,themiceweresubcutaneously(SCgroup) and intranasally (IN group) immunized ,respectively .Four mice from each group were sacrificed in every 2 wk during 0-20 wk after immunization .Splenocyte proliferation was investigated by MTT colorimetric assay ,subsets of CD+4 and CD+8 T cells and apoptosis of splenocytes by FACsort flow cytometry .In SC group ,unstimulated and stimulated with S jAWA ,the level of splenocyte proliferation significantly increased at 4-20 wk after vaccination and increased markedly at 4-18 wk stimulated with ConA ,both of which peaked at 8 wk;in IN group ,the proliferation level of splenocyte cultured with SjAWA and ConA signif-icantly increased during the 4-18 wk ,2-10 wk and 14-18 wk ,2-8 wk and 12-18 wk ,respectively ,and all reached the maximum at the 4 wk after immunization (P＜0 .01 or P＜0 .05) .CD+4 subsets increased obviously during 2-14 wk ,2 wk and 6-16 wk re-spectively ,and reached the peak at 8 wk (P＜0 .01 or P＜0 .05) in both group ,while CD8+ subsets rose lightly during 2-20 wk in both group ,and reached the maximum at 8 wk (SC group) and 6 wk respectively (P>0 .05) .Whether unstimulated or stimulated with ConA ,the level of splenocyte apoptosis of which remarkably increased at 2-4 wk and 2-6 wk separately in SC group ,and both peaked at 2 wk (P＜0 .01 or P＜0 .05 );in IN group ,the level of splenocyte apoptosis all increased at 4 wk and reached the maximum at the same time .In summary ,by inducing the proliferation of splenocytes ,increasing CD4 + T cells and decreasing splenocyte apoptosis ,the rBb (pGEX-Sj26GST ) vaccine plays a critical role in the protective immune response .

Immune stimulatory activity of BRP-4, an acidic polysaccharide from an edible plant, Basella rubra L

Objective: To evaluated the immunomodulatory effect of BRP-4, an acidic polysaccharide from Basella rubra (. B. rubra) L on the macrophage activity. Methods: Phagocytic activity was determined by the ingestion of Latex Beads-Rabbit IgG-FITC using the fluorescent microscopy and flow cytometry analysis and nitric oxide production was measured using Griess reaction assay. Results: An enhanced production of NO was observed at 10 and 100 μg/mL of BRP-4. The phagocytic activity of macrophage was enhanced in BRP-4 treated RAW264.7 cells. BRP-4 combined with concanavalin A (Con A) provided obvious promotion and strengthening of the proliferation of the splenocytes. Conclusions: BRP-4, polysaccharide isolated from B. rubra, is suggested to activate macrophage function and stimulate splenocyte proliferation. The strong immunomodulatory activity of BRP-4 confirmed its good potential as an immunotherapeutic adjuvant.

Effects of Plant Water Extract Codonopsis Lanceolatae on Mouse Immune Cell Activation Ex Vivo

Codonopsis lanceolatae has been used as one of the traditional remedies as well as food source. However, few studies on their immunomodulating effects have been reported. We previously reported that ex vivo supplementation of Codonopsis lanceolatae water extracts enhanced splenocyte proliferation compared to the control group. In order to elucidate its ex vivo effect, six to seven week old balb/c mice were fed ad libitum on a chow diet and water extracts of Codonopsis lanceolatae were orally administrated every other day for four weeks at two different concentrations (50 and 500 mg/kg B.W.). After preparing the single cell suspension, the proliferation of splenocytes was determined by MTT (3-[4,5-dimethylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. The production of cytokine (IL-1beta, IL-6, TNF-alpha), secreted by macrophages stimulated with LPS or not, was detected by ELISA assay using a cytokine kit. After 48 hrs of incubation with the mitogen (ConA or LPS) stimulation, the mice splenocyte proliferation in experimental group was statistically increased at two different concentrations than that in control group. The cytokines production was more significantly enhanced at the lower supplementation (500 mg/kg B.W.) group rather than higher concentration (500 mg/kg B.W.) compared to the control group. The results of this study may suggest that the supplementation of water extract of plant mixture could regulate the immune function by increasing the splenocyte proliferation and enhance the immune function through regulating cytokine production capacity by activated macrophages in mice.

Effects of Plant Water Extract Mixture Ixeris sonchifolia Hance, Oenanthevjavanica, Fagopyrum Esculentum Moench, Hizikia Fusiforme, Zingiber Officinale Roscoe on Mouse Immune Cell Activation Ex vivo

Ixeris sonchifolia Hance (Godulbaegi), Oenanthe javanica (Dolminari), Fagopyrum esculentum Moench (Buckwheat), Hizikia fusiforme (Seaweed Fusiforme) and Zingiber officinale Roscoe (Ginger) have been used respectively as one of folk remedies as well as food materials. However, reportedly few studies on their immunomodulating effects have been made, although it has been known from other preceding studies that the ex vivo supplementation of each Ish, Oj, Fem, Hf, Zor water extracts tends to enhance the proliferation of splenocyte in comparison to the control group. This study on the combined immunomodulative effect of water extract mixture of these five food materials (Ish + Oj + Fem + Hf + Zor) lasted covering seven or eight weeks. The old mice (balb/c) was fed ad libitum on chow diet, and the water extract of plant mixture was orally administrated every other day for four weeks at two different concentrations (50 and 500 mg/kg B.W). After preparing the single cell suspension, the proliferation of splenocyte was determined by MTT (3-[4,5-dimethylthiazol-2-y]-2,5- diphenyl terazolium bromide) assay. The production of cytokine (IL-1beta, IL-6, and TNF-alpha) which was secreted by macrophages stimulated with LPS or not was detected by ELISA assay using the cytokine kit. After the 48 hours of incubation with the mitogen (ConA or LPS) stimulation, the proliferation of the mice splenocyt in the experimental group statisticaly increased at both of two different concentrations in comparison to the control group. The cytokines production was more significantly enhanced at the lower supplementation (50 mg/kg B.W.) group than at the higher concentration (500 mg/kg B.W.). The result of this study may suggest that the supplementation of water extract of plant mixture can regulate and enhance the immune function by increasing the splenocyte proliferation and regulating the cytokine production capacity by the activated macrophages in mice.

Enhancing Effect of Zingiber Officinale Roscoe Extracts on Mouse Spleen and Macrophage Cells Activation

Recently many investigators have initiated searches for immunomodulating substances from natural food sources. Ginger (Zingiber officinale Roscoe) has been used as a raw material in many traditional preparations since the ancient time. This study was performed to investigate the immunomodulative effects of Zingiber officinale Roscoe in mice, using ex vivo experiments. In order to elucidate the immunomodulative effects of Ginger, water extracts of the plant were orally administrated into mice, and isolated splenocytes and macrophages were used as experimental model. In order to identify its ex vivo effect six to seven week old Balb/c mice were fed ad libitum on a chow diet, and water extracts of ginger were orally administrated every other day for two weeks at two different concentrations (50 and 500 mg/kg b.w.). After preparing the single cell suspension, the proliferation of splenocytes was determined by MTT assay. The result of ex vivo study showed that the highest proliferation of splenocytes and macrophage activatation was seen in the mice orally administrated at the concentration of 500 mg/kg b. w. of ginger water extracts. In conclusion, this study suggests that ginger extracts may enhance the immune function by regulating the splenocyte proliferation and cytokine prodution capacity by activated macrophages in mice.

Effects of Scutellaria baicalensis fraction on inflammatory animal model and immune function / 中国药理学通报

Aim To study the anti-inflammatory effects and mechanisms of SBM isolated from Scutellariae radix by high hydrostatic pressure.Methods The effect of SBM on ConA-induced splenocyte proliferation and LPS-induced IL-1? production from peritoneal macrophage was studied in vitro,and the therapeutic effects of SBM on adjuvant-induced arthritis,formalin-induced paw oedema and acetic acid-induced vascular permeability were also investigated.Results SBM(31.25～500 mg?L-1)significantly inhibited ConA-induced splenocyte proliferation,IL-1? production from peritoneal macrophages in vitro.SBM 20 mg?kg-1 inhibited primary and secondary inflammatory reaction,decreased the elevated level of IL-1? released from peritoneal macrophages,inhibited splenocyte proliferation in adjuvant-induced arthritis in rats.Moreover,SBM inhibited formalin-induced paw oedema and acetic acid-induced vascular permeability.Conclusions SBM could inhibit inflammatory reaction.Its mechanisms might be related to the suppression of immune function and inhibition on proinflammatory cytokines production.

Age-dependent changes of B cell and T cell-mediated immune responses in naive C3H/HeJ mice under regular mouse chow feeding conditions / 천식및알레르기

BACKGROUND AND OBJECTIVE: Murine system for studying allergic diseases has been popular in the fields of food allergy and development of their therapeutic strategies. However, there has been no information about the age-dependent changes of natural immune responses of naive C3H/HeJ mice. The purpose of this study was to evaluate the age-dependent changes of B and T-cell mediated immunologic parameters in naive C3H/HeJ mice, which can provide information for experimental planning and analysis of research results. SUBJECTS AND METHODS: Eight naive, female, 5-week-old C3H/HeJ mice were grown under the regular mouse chow feeding conditions for 6 weeks. Sera were obtained at week (w) 5, w6, w8 and w10 for measuring total and chow-specific IgE, IgG1 and IgG2a antibodies. Splenocyte proliferation (at w8 and w10) and cytokine production (at w6, w8 and w10) were evaluated with or without Con A stimulation with pooled splenocytes from two mice of each age group. Serum antibodies and cytokines (IL-4, IL-5, IL-12, INF-gamma, TGF-beta1) were measured by ELISA. Using RT-PCR, IL-4 and INF-gamma mRNA expressions were measured in Peyer's patch and spleen tissue at w10. RESULTS: The levels of total IgE and IgG1 were increased by age while the level of IgG2a was decreased. Chow-specific IgE and IgG2a responses were neglectable through out the whole experimental period (20-30 ng/ml or less). Chow-specific IgG1 levels were measured in the significant concentrations (200-300 ng/ml) but there was no age-dependent change through out the experiment. Con A stimulated-splenocyte proliferation indexes were variable according to the culture-durations and ages of mice. The higher proliferation indexes were observed in the wells receiving thymidine pulse at 48-hour culture, especially in the mice at w10. Con A stimulated IL-4 production in the 72-hour splenocyte culture supernatant was significantly increased at w8, and w10 while INF-gamma production increased only at w10. The changes in the production of IL-5, IL-12 and TGF-beta did not provide significant information in the present study. The ratio of IL-4/IFN-gamma mRNA expression was higher in Peyer's patch than in the spleen. CONCLUSION: The changes of B-cell and T-cell mediated immunologic parameters were complex and variable according to the age in naive C3H/HeJ mice under regular chow feeding conditions. For that reason, the information from the present study needs to be considered in the course of planning or analysing research/data using murine systems.

Effect of vernoniaanthelmintica wild on the immune function of mice / 中国免疫学杂志

Objective:To explore the effect of the vernoniaanthelmintica willd injection on the immune function of mice. Methods: The action of vernoniaanthelmintica willd injection on spleen T, B cell proliferation and antibody production response by [3H]-TdR incorporation and antibody forming cell assays, respectively. Results: Vernoniaanthelmintica willd injection inhibited splenocyte proliferation of normal mouse in vitro and in vivo, the antibody forming cell in vivo, and delayed type of hypersensitivity (DTH) Conclusion: Vernoniaanthelmintica willd injection could inhibit the cellular immune function and humoral immune function.

Isolation and structure elucidation of novel glucan from Astragalus mongholicus / 中草药

Object To isolate the immunoactive polysaccharide from Astragalus mongholicus Bunge and elucidate its chemical structure Methods The polysaccharide was purified from water extracts of A mongholicus by ethanol precipitation, deproteination, selective precipitation with hexadecyltri methylammonium bromide, ion exchange and gel filtration chromatography Its homogeneity and molecular weight were estimated by gel filtration chromatography, the structure was deduced from sugar analysis, methylation analysis, Smith degradation, IR and 13 CNMR spectrophotometry Results A homogeneous polysaccharide A2Nb was obtained with a molecular mass of 360 000 , and composed of D glucose with a major linkage form of ? D (1→4) glucose Side chains were found at 6 O positions once in every 25 glucose residues Conclusion A high molecular weight glucan A2Nb was obtained from A mongholicus for the first time It showed the ability of promoting the proliferation of the splenocytes of mice