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Evaluation of Protective Immune Response Induced by a DNA Vaccine Encoding GRA8 against Acute Toxoplasmosis in a Murine Model

  • Chu, Jia-Qi;Huang, Shuai;Ye, Wei;Fan, Xuan-Yan;Huang, Rui;Ye, Shi-Cai;Yu, Cai-Yuan;Wu, Wei-Yun;Zhou, Yu;Zhou, Wei;Lee, Young-Ha;Quan, Juan-Hua
    • Parasites, Hosts and Diseases
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    • v.56 no.4
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    • pp.325-334
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    • 2018
  • Toxoplasma gondii is an apicomplexan zoonotic protozoan parasite that infects most species of warm-blooded animals, including humans. The heavy incidence and severe or lethal damage caused by T. gondii infection clearly indicate a need for the development of an effective vaccine. T. gondii GRA8 is a member of the dense granules protein family and is used as a marker of acute infection. In the present study, we evaluated the protective immunity induced by DNA vaccination based on a recombinant eukaryotic plasmid, pDsRed2-GRA8, against acute toxoplasmosis in mice. BALB/c mice were intramuscularly immunized with the pDsRed2-GRA8 plasmid and then challenged by infection with the highly virulent GFP-RH strain of T. gondii. The specific immune responses and protective efficacy against T. gondii of this vaccine were analyzed by measuring cytokine and serum antibody titers, splenocyte proliferation assays, and the survival times of mice after challenge. Our results showed that mice immunized with pDsRed2-GRA8 demonstrated specific humoral and cellular responses, induced higher IgG antibody titers with predominant IgG2a production; increased levels of IL-10, IL-12 (p70), $IFN-{\gamma}$, $TNF-{\alpha}$, and splenocyte proliferation; and prolonged survival times compared to those of control mice. The present study showed that DNA immunization with pDsRed2-GRA8 induced humoral and cellular immune responses, and all immunized mice showed greater Th1-type immune responses and longer survival times than those of control mice. These results indicated that T. gondii GRA8 DNA immunization induces a partial protective effect against acute toxoplasmosis.

The Influence of Estrogen on the Immunologic Function of T Lymphocytes (에스트로겐이 T 림프구 면역기능에 미치는 영향)

  • Chun, Sung-Wook;Chun, Kyung-Chun;Ku, Seung-Yup
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.2
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    • pp.89-100
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    • 2009
  • SLE 등의 자가면역질환 병인론에 있어서 $CD4^+$ T 조력 림프구 (T helper cell)의 기능과 항상성 조절의 이상이 중요하다는 것은 알려진 사실이지만 $CD4^+$ T 조력 림프구 (T helper cell)의 발달과 기능에 대한 성호르몬의 정확한 역할에 대하여는 아직 확립되어 있지 않은 실정이다. 에스트로겐의 $CD4^+$ T 림프구에 대한 여러 방면의 작용에 대한 결과는 에스트로겐 매개 신호가 전달되는 정황에 매우 의존적이라고 할 수 있으며, 또한 에스트로겐이 B 림프구와 항원발현세포들 같이 T 림프구 기능을 변화시킬 수 있는 다른 분획 세포들의 기능에 대해서도 영향을 미칠 수 있다는 사실을 고려한다면, 에스트로겐이 $CD4^+$ T 림프구매개반응에 미치는 작용은 매우 복잡하다고 할 수 있다. 현재로선 이런 과정들과 관련된 에스트로겐의 정확한 기전에 대한 연구 성과는 현재로선 아직걸음마 단계에 지나지 않는다고 볼 수 있다. 일부 분자학적 표적들의 발견에도 불구하고 ER-$\alpha$와 ER-$\beta$, 또는 이런 표적들의 조절에 대한 유전자적 또는 비유전자적 경로의 상대적 역할에 대해서는 현재로선 알려진 바가 많지 않은 것 또한 사실이며, T 림프구 자체의 미세 환경과 ER 매개 전사반응에 영향을 미칠 수 있는 방법 역시 아직까진 알려진 바가 없다. 따라서 에스트로겐의 $CD4^+$ T 림프구에 대한 다각적 영향을 규명하는 데 있어 이런 경로들에 대한 보다 체계적이고 상세한 연구가 반드시 필요하다. 최근 골다공증의 병인에 있어 에스트로겐에 의한 TNF 및 RANK-RANKL계 억제에 대해 연구가 심도 있게 진행됨으로써 결과적으로 다른 시토카인과 면역세포들에 대한 영향이 간접적으로 규명되고 있는 점은 매우 고무적인 현상이라고 여겨진다. 이러한 연구들은 추후 에스트로겐의 면역 및 염증체계에 대한 특징적 작용을 규명하는데 있어서 밑거름이 될 수 있을 것이다. 나아가 호르몬 자체, 또는 SERM 같은 준호르몬제제들이 면역계에 미치는 영향을 분자학적으로 규명함으로써 향후 이러한 지식들이 또한 류마티스성 자가면역질환이나 만성염증성질환, 또는 전신성 감염질환에 있어 면역글로불린이나 기타 다른 기존 치료 약제들을 대체 보완할 수 있는 호르몬 치료로 이어질 수 있는 결정적인 단서를 제공할 수 있을 것이라 기대할 수 있을 것이다.

Rare Helicobacter pylori Infection May Explain Low Stomach Cancer Incidence: Ecological Observations in Bali, Indonesia

  • Tanaka, Tsutomu;Mulyadi, I Ketut;Moestikaningsih, Moestikaningsih;Oka, Tjok Gede;Soeripto, Soeripto;Triningsih, FX Ediati;Triyono, Teguh;Heriyanto, Didik Setyo;Hosono, Akihiro;Suzuki, Sadao;Tokudome, Shinkan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.979-984
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    • 2016
  • The incidence rate of stomach cancer in Bali, Indonesia, is estimated to be strikingly lower than that in Japan. We conducted an on-site ecological study to investigate the association between the stomach cancer incidence and Helicobacter pylori (H. pylori) infection. Recruiting 291 healthy persons (136 men and 155 women) from the general population in Bali, Indonesia, we conducted a urea breath test (UBT) to examine H. pylori infection, along with a pepsinogen test to detect chronic atrophic gastritis and urine analysis to estimate sodium and potassium excretion. UBT positivities were 9% (2-15, 95% confidence interval) for men and 7% (1-12) for women, and positive cases for H. pylori IgG antibodies were 1% (0-3) for men and 3% (0-5) for women, significantly lower than the respective values in Japan. Positive pepsinogen tests in Bali were 0% (0-0) for men and 1% (0-4) for women, also significantly lower than the Japanese figures. Computed values for daily salt excretion were $13.3{\pm}4.1g$ (mean${\pm}$SD) for men and $11.1{\pm}3.1g$ for women, as high as corresponding Japanese consumption values. Moreover, the estimated potassium excretion was $3.2{\pm}0.7g$ for men and $2.8{\pm}0.6g$ for women in Bali, significantly higher than the figures in Japan. There were no associations across genetic polymorphisms of IL-beta, TNF-alpha, and PTPN11 with UBT positivity. The low incidence of stomach cancer in Bali may thus mainly be due to the rare H. pylori infection. Namely, the bacterium infection seems to be a critical factor for gastric cancer rather than host or other environmental factors.

The Effects of 12 Weeks Combined Exercise on Brain Nerve Growth Factor, Inflammation-Related Factor in Obese High School Girls (12주 복합운동이 비만 여고생의 뇌신경세포 생성인자 및 염증인자에 미치는 영향)

  • Seo, Jeongpyo;Heo, Junhoe;Kim, Hyunjun;Park, Jangjun
    • Journal of The Korean Society of Integrative Medicine
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    • v.8 no.1
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    • pp.159-168
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    • 2020
  • Purpose : To provide data on exercise prescription for obesity management and prevention of cardiovascular disease in girl's high school and to prepare basic data for more effective exercise program for lifestyle improvement and prevention of lifestyle-related diseases. This study examines the effects on brain nerve growth factor and inflammatory factors, and the relationship between obesity factor and brain neuron cell production factor and inflammatory factor changes by complex exercise. Methods : The subjects of the study were obese students with a body fat percentage of 30 % or higher after obtaining body fat percentage of high school girls in C-city. Among them, 20 students who wanted to participate in the program of this study and did not participate in special exercise and diet therapy within the last 6 months were radio-sampled into groups of exercise group and control group, but attendance rate was low and The final exercise group (9) and control group (9) were measured, except for one student who did not respond. Results : Analysis of the range of variation in body composition, BMI, lean body mass, and the interaction between the groups showed significant differences (p<.05). TC, TG, HDL-C, and LDL-C as variables of blood lipids, TC and TG were not significantly different and TG was significantly different (p<.05) in interactions. HDL-C showed a significant difference (p<.01) in interactions, an increase in exercise group, and a significant decrease in control group (p<.05). There was a significant difference (p<.05) in BDNF interaction, an increase in the exercise group and a decrease in the control group, but no significant difference. NGF tended to increase in both exercise and control groups. IL-6 had a significant difference in timing (p<.05) and significantly decreased (p<.01) in the exercise group, and TNF-α interacted with timing (p<.05), and a significant increase in the control group. Conclusion : This study confirmed 12-week compound exercise program was effective in increasing the expression of basal fitness or CNS factor, but not enough to actually improve brain function. Fat mass and obesity are also affecting vascular inflammatory factors.

Metformin or α-Lipoic Acid Attenuate Inflammatory Response and NLRP3 Inflammasome in BV-2 Microglial Cells (BV-2 미세아교세포에서 메트포르민 또는 알파-리포산의 염증반응과 NLRP3 인플라마솜 약화에 관한 연구)

  • Choi, Hye-Rim;Ha, Ji Sun;Kim, In Sik;Yang, Seung-Ju
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.253-260
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    • 2020
  • Alzheimer's disease (AD) is a chronic and progressive neurodegenerative disease that can be described by the occurrence of dementia due to a decline in cognitive function. The disease is characterized by the formation of extracellular and intracellular amyloid plaques. Amyloid beta (Aβ) is a hallmark of AD, and microglia can be activated in the presence of Aβ. Activated microglia secrete pro-inflammatory cytokines. Furthermore, S100A9 is an important innate immunity pro-inflammatory contributor in inflammation and a potential contributor to AD. This study examined the effects of metformin and α-LA on the inflammatory response and NLRP3 inflammasome activation in Aβ- and S100A9-induced BV-2 microglial cells. Metformin and α-LA attenuated inflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). In addition, metformin and α-LA inhibited the phosphorylation of JNK, ERK, and p38. They activated the nuclear factor kappa B (NF-κB) pathway and the NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome. Moreover, metformin and α-LA reduced the marker levels of the M1 phenotype, ICAM1, whereas the M2 phenotype, ARG1, was increased. These findings suggest that metformin and α-LA are therapeutic agents against the Aβ- and S100A9-induced neuroinflammatory responses.

Possibility of Involvement of Porphyromonas gingivalis in Coronary Heart Disease

  • Lee, Jin-Yong;Park, Byung-Lae;Yun, Hyun-Kyung;Park, Eun-Ah;Shin, Eun-Ah;Jue, Seong-Suk;Shin, Je-Won
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.203-214
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    • 2000
  • Porphyromonas gingivalis has been implicated in periodontal diseases. Accumulating evidence suggests that cardiovascular disease is the most prevalent medical problem in patients with periodontal diseases. In order to check the possibility that P. gingivalis is involved in coronary heart disease, the present study was performed to observe P. gingivalis adherence and invasion of human coronary artery endothelial cells (HCAEC) and production of cytokines and growth factors by HCAEC upon P. gingivalis infection. $^3H$-labeled P. gingivalis 381 was incubated with HCAEC for 90 min. The radioactivity of the washed HCAEC was a measure of the absorbed (adhering and invading) P. gingivalis. The absorption radioactivity of the HCAEC infected by P. gingivalis was determined to be 59.58% of the input bacterial cells. In contrast, the absorption radioactivity of the cells infected by S. gordonii Challis which was employed as a control was negligible (0.59%). DPG3, a P. gingivalis mutant defective of fimbriae, appeared to be impaired to some extent in capability of adherence/invasion as compared to that of the parental strain 381, showing 43.04% of the absorption radioactivity. The absorption radioactivity of the HCAEC infected by P. gingivalis 381 in the presence of excessive fimbriae at the concentrations of $50\;{\mu}g$ and $100\;{\mu}g/ml$ was 57.27 and 45.44%, respectively. Invasion of HCAEC by P. gingivalis 381 was observed by an antibiotic (metronidazole) protection assay and transmission electron microscopy (TEM). In the antibiotic protection assay, invasion by the bacterium was measured to be 0.73, 1.09, and 1.51% of the input bacterial cells after incubation for 30, 60, and 90 min, respectively. Invasion by DPG3 was shown to be 0.16% after 90-min incubation. In comparison of invasion efficiency at 90 min of the incubation, the invasion efficiency of DPG3 was 0.37% while that of its parental strain 381 was 2.54%. The immunoblot analysis revealed fimbriae of P. gingivalis did not interact with the surface of HCAEC. These results suggest that fimbriae are not the major contribution to the adherence of P. gingivalis to HCAEC but may be important in the invasion of HCAEC by the bacterium. The presence of cytochalasin D ($1\;{\mu}g/ml$) and staurosporine ($1\;{\mu}M$) reduced the invasion of HCAEC by P. gingivalis 381 by 78.86 and 53.76%, respectively, indicating that cytoskeletal rearrangement and protein kinase of HCAEC are essential for the invasion. Infection of P. gingivalis induced HCAEC to increase the production of TNF-${\alpha}$. by 60.6%. At 90 min of the incubation, the HCAEC infected with P. gingivalis cells was apparently atypical in the shape, showing loss of the nuclear membrane and subcellular organelles. The overall results suggest that P. gingivalis may cause coronary heart disease by adhering to and invading endothelial cells, and subsequently damaging the cells.

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Effects of the immobilization of heparin and rhPDGF-BB to titanium surfaces for the enhancement of osteoblastic functions and anti-inflammation

  • Huh, Jung-Bo;Lee, Jeong-Yo;Lee, Kyung-Lae;Kim, Sung-Eun;Yun, Mi-Jung;Shim, Ji-Suk;Shim, June-Sung;Shin, Sang-Wan
    • The Journal of Advanced Prosthodontics
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    • v.3 no.3
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    • pp.152-160
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    • 2011
  • PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

An Experimental Study on the Biological Specificity of Nitric Oxide and Nitric Oxide Synthetase in Periodontium-Related Cells (치주조직 유관세포에서의 Nitric Oxide 및 Nitric Oxide Synthetase의 생물학적 특성에 관한 연구)

  • Yoon, Hyung-Jin;Yoon, Dong-Whan;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.27 no.4
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    • pp.883-908
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    • 1997
  • Bone remodeling is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. The process is tightly regualted at the local level by an incompletely known netwotk of peptide and non-peptide fators. Nitric oxide(NO), synthesized by nitric oxide synthetase(NOS) from L-arginine, is becoming recognized as an important bioregualtory molecule in a variety of tissue, but little is known about its possible role in periodontal tissue. The purpose of this study is to investigate the expression of nitric oxide synthetase(NOS) in inflamed gingiva and the effects of cytokine on the expression of NOS protein. The expression of NOS in gingival tissue was evaluated by immunohistochemical staining for $NOS_1$, $NOS_2$, $NOS_3$. The effect of cytokine on the expression of NOS in human periodontal ligament cells and osteoblast-like HOS cells by western blot analysis. Further, we studied that NO functions in periodontal ligament cells as a regulatory molecule. PDL cells incubated with NOS inhibitor and donor. The protein expression, type I collagen & non-collagenous protein, nitrate production and cell proliferation were evaluated The results were as follows. 1. $NOS_1$, $NOS_2$, $NOS_3$ was rarely distributed in healthy gingiva, but stronger stained in gingival epithelium, endothelial cells, and mononuclear cells of inflammed gingiva. 2. The cytokine stimulated $NOS_1$, and $NOS_3$ protein were not inducing or inhibitory effect to compared with control in PDL and HOS cells. 3.Incubation of cells with combination of $TNF-{\alpha}$, $IFN-{\gamma}$, LPS result in a time dependant increase in $NOS_2$ expression, reaching a maximal level after 24 hours of stimulation. 4. The osteonectin protein inhibitory effect of NMA, inhibitor of NOS, was reversed by Larginine in dose dependant manner. 5. NMA decreased cell poliferation and nitrate production, but the inhibitory efffect of NMA was also prevented by the NO donor, sodium nitropruiside. These results suggest that exogenously synthesized NO was playing a stimulating effect on cell proliferation or on non-collagenous protein expression. Therefore NO have an important role in mediation of localized bone destruction associated inflammatory bone disease such as periodontitis.

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Effect of Green Tea Probiotics on the Growth Performance, Meat Quality and Immune Response in Finishing Pigs

  • Ko, S.Y.;Yang, C.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.9
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    • pp.1339-1347
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    • 2008
  • The objective of this study was to determine the effects of green tea probiotics on growth performance, meat quality and immune response in finishing pigs, and to assess the possibility of substituting green tea probiotics for antibiotics in diets of finishing pigs. This green tea probiotics is made by mixing green tea powder and excipients (defatted rice bran and wheat bran) and fermenting the mixture with beneficial bacteria. A total of 90 crossbreed "Landrace$\times$Yorkshire" finishing pigs with an average body weight of $72.5{\pm}2.5kg$ were assigned to 5 dietary treatments in a completely randomized design. Each treatment had 3 replications with 6 pigs per replication. The five dietary treatments were control, antibiotic (0.003% chlortetracycline added) and 0.1, 0.5 and 1.0% of green tea probiotics. There were no significant differences in final body weight, daily weight gain, daily feed intake and feed conversion ratio in the green tea probiotics and antibiotic treatments (p>0.05). Crude protein content was significantly increased in the 0.1 and 1.0% green tea probiotics treatment groups (p<0.05) and there was no significant difference in crude fat content of the meat among the treatments. The TBA value of meat was significantly lowered with 0.5 and 1.0% green tea probiotics treatments compared to that of controls and statistically similar to the antibiotic treatment after 3 weeks of storage (p<0.05). The growth of spleen cells stimulated with Con A (0.1 and $1.0{\mu}g/ml$) was significantly increased with 1.0% green tea probiotics treatment compared to that of the control treatment (p<0.05). The growth of spleen cells stimulated with LPS (1.0, 3.0 and $10{\mu}g/ml$) was significantly increased in the 0.5% green tea probiotics group compared to the antibiotic group (p<0.05). In Con A ($1.0{\mu}g/ml$) medium, IL-6 production of spleen cells was significantly increased with 1.0% green tea probiotics treatment compared to that of the control (p<0.05). In LPS ($10.0{\mu}g/ml$) medium, TNF-${\alpha}$ production of spleen cells increased significantly in all green tea probiotics treatment groups compared to that of the control (p<0.05). Finally it can be summarized that addition of green tea probiotic has a positive effect similar to antibiotic and 0.5% is the suitable dietary supplementation dose for finishing pig production.

Immunomodulatory Activity of Crude Polysaccharide Separated from Cudrania tricuspidata Leaf (꾸지뽕(Cudrania tricuspidata) 잎으로부터 분리된 다당류 추출물의 면역 활성)

  • Byun, Eui-Baek;Jang, Beom-Su;Sung, Nak-Yun;Byun, Eui-Hong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.8
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    • pp.1099-1106
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    • 2016
  • The objective of this study was to evaluate the immunomodulatory activity of crude polysaccharide separated from Cudrania tricuspidata leaf. C. tricuspidata polysaccharide (CTP) was extracted by ethanol precipitation. Immunomodulation activity was tested in macrophage cells (RAW 264.7 and bone-marrow derived macrophage) and splenocytes. CTP treatment significantly increased cell proliferation up to $250{\mu}g/mL$ in both RAW 264.7 and bone-marrow derived macrophages. In this concentration range (below $250{\mu}g/mL$), nitric oxide and cytokine [tumor necrosis factor $(TNF)-{\alpha}$ and interleukin (IL)-6] production also significantly increased. Similarly, splenocyte proliferation dosedependently increased except for the $1,000{\mu}g/mL$ treated group. Regarding cytokine production activity in splenocytes, CTP treatment significantly increased production of Th 1 type cytokines [interferon $(IFN)-{\gamma}$] production but not Th 2 type cytokines (IL-4). Therefore, the results indicate that CTP may have a potential effect on immunomodulatory activity in various immune cells, and this is useful for development of immune enhancing adjuvant materials as a natural ingredient.