• Title/Summary/Keyword: splenocytes

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Immunogenicity of a DNA and Recombinant Protein Vaccine Combining LipL32 and Loa22 for Leptospirosis Using Chitosan as a Delivery System

  • Umthong, Supawadee;Buaklin, Arun;Jacquet, Alain;Sangjun, Noppadol;Kerdkaew, Ruthairat;Patarakul, Kanitha;Palaga, Tanapat
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.526-536
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    • 2015
  • Leptospirosis is a worldwide zoonotic disease caused by pathogenic Leptospira, a genus of which more than 250 serovars have been identified. Commercial bacterin vaccines are limited in that they lack both cross-protection against heterologous serovars and long-term protection. This study investigated in mice the immunogenicity of an anti-leptospirosis vaccine, using the outer membrane proteins LipL32 and Loa22 as antigens. The immunogenicity of this vaccine formulation was compared with those induced by vaccines based on LipL32 or Loa22 alone. A DNA-encapsulated chitosan nanoparticle was used for in vivo DNA delivery. Using a unique DNA plasmid expressing both lipL32 and loa22 for vaccination, higher antibody responses were induced than when combining plasmids harboring each gene separately. Therefore, this formulation was used to test the immunogenicity when administered by a heterologous prime (DNA)-boost (protein) immunization regimen. The specific antibody responses against LipL32 (total IgG and IgG1) and Loa22 (IgG1) were higher in mice receiving two antigens in combination than in those vaccinated with a single antigen alone. Although no significant difference in splenic CD4+ T cell proliferation was observed among all groups of vaccinated mice, splenocytes from mice vaccinated with two antigens exhibited higher interferon-γ and IL-2 production than when using single antigens alone upon in vitro restimulation. Taken together, the immunogenicity induced by LipL32 and Loa22 antigens in a heterologous primeboost immunization regimen using chitosan as a DNA delivery system induces higher immune response, and may be useful for developing a better vaccine for leptospirosis.

Immune-enhancing effect of Acanthopanax Koreanum and its component, Eleutheroside E on the protein-energy malnourished C57bl/6 mice

  • Kim, Na-Hyung;Kim, Kyu-Yeob;Kim, Jeong-Ah;Kim, Young-Ho;Kang, In-Cheol;Kim, Hyung-Min;Jeong, Hyun-Ja
    • Advances in Traditional Medicine
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    • v.10 no.3
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    • pp.191-199
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    • 2010
  • Acanthopanax Koreanum stem (AK) has been used in Korea as a tonic and sedative as well as a drug with ginseng like activities. The purpose of our present study was to investigate the effects of AK extract (AKE) and Eleutheroside E, major component of AKE on an exacerbated immune function through utilization of protein-energy malnutrition (PEM) diet by using forced swimming test (FST). The immobility time were significantly decreased in the AKE or Eleutheroside E-administrated group compared with the control group on the FST (P < 0.05). The level of blood parameters were not changed significantly. PEM-induced weight loss of mice was reduced by oral administration of 500 mg/kg AKE. AKE oral administration improved the nutritional status such as the food efficiency ratio and the adrenal gland weight. AKE treatment significantly increased the production of interferon (IFN)-$\gamma$ compared with unstimulated splenocytes but not interleukin (IL)-4. Eleutheroside E also significantly increased the IFN-$\gamma$ production but not IL-2 and IL-4 in T cell line, MOLT-4 cells. These results suggest that AKE and Eleutheroside E may influence to immune-enhancing through increasing the physical endurance capacity and immune cell activation.

Effects of Seokgokamibokhapbang(SGBHB) on anti-CD40-and recombinant IL-4-induced cytokine production and immunoglobulin E and histanine release in highly purified mouse B cells (석고가미복합방이 Anti-CD40과 rIL-4로 유도된 생쥐의 B 세포에서 싸이토카인 생성 및 면역글로블린 E에 미치는 효과)

  • Choi, Moon-Suk;Kim, Su-Myung;Namgung, Uk;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.13 no.2
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    • pp.131-146
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    • 2004
  • In the present study, we exarnined anti-allergic effect of SGBHB in cultured B cells. B cells were prepared from isolated murine splenocytes and activated by co-treatment of anti-CD40 monoclonal antibody and recombinant IL-4 allergens. Anti-allergic effects of SGBHB in activated B cells were determined by measuring B cell surface activated molecules (CD23+ and CD11a+), and expression levels of IL-$1{\beta}$, IL-6, IL-10, TNF-$\alpha$, IgE, and HRF. The major findings are summarized as follows. 1. SGBHB treatment did not produce significant cytotoxic effects on mouse lung fibroblast cells. 2. SGBHB produced significant inhibitory effect on the expression of B cell surface activated molecules (CD23+ and CD11a) in activated B cells. 3. SGBHB treatment significantly inhibited expression levels of IL-$1{\beta}$, IL-6, and TNF-$\alpha$ mRNAs in activated B cells.IL-6 protein levels were significantly decreased by $100{\mu}g/m{\ell}$ of SGBHB treatrrient, and TNF-$\alpha$ protein levels were decreased compared to the control group, but statistically insignificant. 4. SGBHB treatment significantly increased IL-10 at both mRNA and protein levels in activated B cells. 5. SGBHB treatment significantly inhibited levels of IgE production. Thus, the present data suggest that SGBHB has an anti-allergic effect on activated B cells by controlling irnmune responses, and further implicates the possibility on clinical application as a therapeutic agent.

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Dehydroepiandrosterone Sulfate Inhibited Immune Dysfunction Induced by LP-BM5 Leukemia Retrovirus Infection through Regulating Th1/Th2 Type Cytokine mRNA Expression and Oxidative Stress in Murine AIDS Model (LP-BM5 Leukemia Retrovirus 감염으로 유발된 Murine AIDS에서 Dehydroepiandrosterone Sulfate의 Th1/Th2 Cytokines의 발현 조절 및 산화적 스트레스 억제 효과 연구)

  • Lee, Jeong-Min
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1329-1335
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    • 2006
  • LP-BM5 murine leukemia retrovirus induces the excessive oxidative stress and immune dysfunction leading to B cell leukemia and murine AIDS with cytokine dysfunction. In the present study, the immune restoratory effect of antioxidant hormone dedydroepiandrosterone sulfate (DHEAS) was investigated in the primary splenocytes from LP-BM5 retrovirus-infected C57BL/6 mice. DHEAS significantly increased T and B cell response to mitogen and normalized the unbalanced production of Th1/Th2 type cytokines. In particular, both protein and mRNA expression of IL-4, IL-6, and $TNF-\alpha$ were down-regulated by DHEAS treatment whereas IL-2 and $IFN-\gamma$ level were increased. This result suggests that DHEAS directly or indirectly regulates the gene expression of Th1/Th2 type cytokines in transcription level. In addition, DHEAS treatment decreased the hepatic lipid peroxidation and preserved vitamin E level in liver cells. These results suggested that DHEAS could effectively prevent immune dysfunction by regulating cytokine secretion and preventing the oxidative stress in murine AIDS.

Effect of Pine needle Ethanol Extracts on the Inhibitory Activity of Atopic Dermatitis (송엽 에탄올 추출물의 아토피 저해 활성)

  • Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Jung, Seul-A;Kim, Hyun-Jee;Kang, Bo-Kyeong;Bark, Si-Woo;Kim, Tae-Wan;Ahn, Dong-Hyun
    • KSBB Journal
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    • v.28 no.2
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    • pp.123-130
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    • 2013
  • The aim of this study was to examine inhibitory effects of pine needle ethanol extracts (PNEE) on atopic dermatitis (AD). To determine inflammatory activity PNEE was added to LPS-induced murine peritoneal macrophages for an in-vitro test. In addition, anti-AD test was carried out by spreading PNEE on the dorsal skin of 2,4-dinitrochlorobenzene (DNCB)- induced BALB/c mice. It was confirmed that the nitric oxide (NO) secretion was suppressed when $1{\sim}50{\mu}g/mL$ of PNEE were added to LPS-induced murine peritoneal macrophages. Moreover, levels of TNF-${\alpha}$, IL-6, and IL-$1{\beta}$, were decreased. For the anti-AD test, PNEE alleviated symptoms of the erythema in DNCB-induced mice. Furthermore, the IFN-${\gamma}$ secretion of the group treated with PNEE was increased in splenocytes from DNCB-induced mice compared to the positive control, while IL-4 secretion diminished. Through these results, we can conclude that PNEE can inhibit AD by modulating the IFN-${\gamma}$, IL-4 cytokines production and inhibiting inflammation.

Study on Antitumor Activity and Radioprotective effects of Kamisasammaekmundongtang (가미사삼맥문동탕(加味沙蔘麥門冬湯)이 항암활성(抗癌活性)과 방사선부작용(放射線副作用) 억제효과(抑制效果)에 미치는 영향(影響))

  • Park, Yang-chun;Kim, Byeong-tak
    • Journal of Haehwa Medicine
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    • v.8 no.1
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    • pp.403-424
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    • 1999
  • To evaluate the antitumor activity, antimetastatic and radioprotective effects of Kamisasammaekmundongtang(KSMT), studies were done experimentally. The results were obtained as follows: 1. In cytotoxicity against P388, A549 and B16-F10, KSMT was not showed satisfiable cytotoxicity as compared with control. 2. In Inhibitory effect on activity of DNA topoisomerase I, KSMT has strong inhibitory effect. 3. The inhibitory effect on adhesion of A549 to complex extracellular matrix was significantly increased at 0.5mg/ml, 1mg/ml of KSMT. 4. The T/C% was 122 in KSMT treated group in S-180 bearing ICR mice. 5. In antiangiogenetic effect on CAM assay, inhibitory rate was 33% in KSMT treated group. 6. In pulmonary colonization assay, a number of colonies in the lungs were decreased significantly in KSMT treated group as compared with control group. 7. By FACS analysis of splenic leukocyte after exposure to radiation by linear accelerator, T-helper cell, B cell and macrophage in KSMT treated group were significantly increased while splenocytes were decreased in control group. 8. In histological changes of jejunum of $Bald{\setminus}C$ mice after exposure to radiation by linear accelerator, exclusion and fusion of villi were decreased as compared with control group. But in duodenum and ileum, exclusion and fusion of villi were not decreased as compared with control group. 9. WBC, PLT were increased in KSMT treated group as compared with control group after exposure to radiation by linear accelerator, but the increasing effect was not significant. Above results suggest that KSMT may be useful in prevention of cancer metastasis and protection from damage by radiotherapy. But the further study of KSMT would be demanded.

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Jaundice in Cultured Eel (Anguilla japonica) (양식 뱀장어 (Anguilla japonica)의 황달에 관하여)

  • KIM, Dae-Hyun;PARK, Sung-Woo
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.6
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    • pp.1573-1580
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    • 2016
  • The disease occurred in cultured eel (Anguilla japonica) in a recirculatory culture system without any separated filtration apparatus. As the pond had a high level of nitrite with $60mg/{\ell}$, 1% NaCl was added to reduce nitrite toxicity to eel. The first outbreak was observed a week after the NaCl treatment and continued for 10 days. Accumulated mortality was about 0.2-0.5%. Affected fish ranged from 150-350 g were usually anorexic and exhibited yellow colour in the skin of the abdominal region and at the base of pectoral fins, as well as in the eyes. In a few individuals with severe symptoms, the lateral skin was also yellowish. The spleen, kidney, muscle and gall bladder were yellowish and the liver was pale-yellow colour but green on the posterior part. The gall bladder was shrunken without bile. Some abnormal erythrocytes such as "tear drop" cells (dacrocyte) were observed in peripheral blood smears stained with May-Grunwald Giemsa. Hematocrit values and hemoglobin contents in the jaundiced eel were significantly lower compared with apparently heathy eel. Severe haemosiderosis accompanied by erythrophagocytosis was found in the kidney and spleen. Haemosiderin deposits were observed in macrophages of the haematopoietic tissue of the kidney and in the splenocytes. But no significant alterations were found in the hepatic cells. In this study we report the first outbreak of jaundice in cultured eel in Korea. Pathological and hematological investigations suggested that severe hemolysis may resulted in jaundice in eel although the cause of hemolytic jaundice was not identified in this study.

Ginsenoside Rp1 Exerts Anti-inflammatory Effects via Activation of Dendritic Cells and Regulatory T Cells

  • Bae, Jin-Gyu;Koo, Ji-Hye;Kim, Soo-Chan;Park, Tae-Yoon;Kim, Mi-Yeon
    • Journal of Ginseng Research
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    • v.36 no.4
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    • pp.375-382
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    • 2012
  • Ginsenoside Rp1 (G-Rp1) is a saponin derivate that provides anti-metastatic activities through inhibition of the NF-${\kappa}B$ pathway. In this study, we examined the effects of G-Rp1 on regulatory T cell (Treg) activation. After treatment of splenocytes with G-Rp1, Tregs exhibited upregulation of IL-10 expression, and along with dendritic cells (DCs), these Tregs showed increased cell number compared to other cell populations. The effect of G-Rp1 on Treg number was augmented in the presence of lipopolysaccharide (LPS), which mimics pathological changes that occur during inflammation. However, depletion of DCs prevented the increase in Treg number in the presence of G-Rp1 and/or LPS. In addition, G-Rp1 promoted the differentiation of the memory types of $CD4^+Foxp3^+CD62L^{low}$ Tregs rather than the generation of new Tregs. In vivo experiments also demonstrated that Tregs and DCs from mice that were fed G-Rp1 for 7 d and then injected with LPS exhibited increased activation compared with those from mice that were injected with LPS alone. Expression of TGF-${\beta}$ and CTLA4 in Tregs was increased, and upregulation of IL-2 and CD80/CD86 expression by DCs affected the suppressive function of Tregs through IL-2 receptors and CTLA4. These data demonstrate that G-Rp1 exerts anti-inflammatory effects by activating Tregs in vitro and in vivo.

Effect of Glycyrrhizae Radix on the Immune Responses(I) - Immuno-regulatory Action of 50% Methanol Extract - (감초가 면역반응에 미치는 영향 (I) - 50% 메탄올 엑스의 면역조절작용 -)

  • 한종현;오찬호;은재순
    • YAKHAK HOEJI
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    • v.35 no.3
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    • pp.154-164
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    • 1991
  • These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

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Effects of GP extract on oxidative stress and contact dermatitis in NC/Nga Mice induced by DNCB (가감평위산(加減平胃散)이 산화적 손상과 접촉성 피부염에서의 면역 조절작용에 미치는 영향)

  • Park, Eung-Ho;Yun, Mi-Young;Kim, Seon-Bin;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.16 no.2
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    • pp.131-145
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    • 2007
  • To evaluate the effects of GP on contact dermatitis, we examined the composition of immune cells from drain lymph node in DNCB-induced contact dermatitis murine model NC/Nga mice. And the amount of pathologic cytokines of spleen and antioxidant activity were investigated. The results were summarized as followers; 1. GP did not show cytotoxic effect on mLFC in vitro. 2. GP did not have hepatotoxicity in vivo in the level of ALT, AST. 3. GP decreased the production of DPPH and in a dose-dependent. 4. GP significantly decreased total cell number of DLN in DNCB-induced NC/Nga mice compared to the untreated control group. 5. GP significantly decreased the number of CD3+, CD19+, CD4+, CD8+, CD3+/CD69+ and CD4+/CD45+ in DLN of DNCB-induced NC/Nga mice compared to the untreated control group. 6. GP significantly reduced the level of IL-4 and IFN-$\gamma$ in splenocytes of DNCB-induced NC/Nga mice compared to the untreated control group. Taken together above results, GP have therapeutic effects on contact dermatitis by regulating T cell activation. This study warranted further investigations of molecular mechanisms of GP on contact dermatitis.

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