• Title/Summary/Keyword: splenocytes

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A Novel Recombinant BCG Vaccine Encoding Eimeria tenella Rhomboid and Chicken IL-2 Induces Protective Immunity Against Coccidiosis

  • Wang, Qiuyue;Chen, Lifeng;Li, Jianhua;Zheng, Jun;Cai, Ning;Gong, Pengtao;Li, Shuhong;Li, He;Zhang, Xichen
    • Parasites, Hosts and Diseases
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    • v.52 no.3
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    • pp.251-256
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    • 2014
  • A novel recombinant Bacille Calmette-Guerin (rBCG) vaccine co-expressed Eimeria tenella rhomboid and cytokine chicken IL-2 (chIL-2) was constructed, and its efficacy against E. tenella challenge was observed. The rhomboid gene of E. tenella and chIL-2 gene were subcloned into integrative expression vector pMV361, producing vaccines rBCG pMV361-rho and pMV361-rho-IL2. Animal experiment via intranasal and subcutaneous route in chickens was carried out to evaluate the immune efficacy of the vaccines. The results indicated that these rBCG vaccines could obviously alleviate cacal lesions and oocyst output. Intranasal immunization with pMV361-rho and pMV361-rho-IL2 elicited better protective immunity against E. tenella than subcutaneous immunization. Splenocytes from chickens immunized with either rBCG pMV361-rho and pMV361-rho-IL2 had increased $CD4^+$ and $CD8^+$ cell production. Our data indicate recombinant BCG is able to impart partial protection against E. tenella challenge and co-expression of cytokine with antigen was an effective strategy to improve vaccine immunity.

Inhibitory Effect of Electroacupuncture on Murine Collagen Arthritis and its Possible Mechanisms

  • Fang, Jian-Qiao;Aoki, Eri;Yu, Ying;Sohma, Toshimitsu;Kasahara, Takako;Hisamitsu, Tadashi
    • Journal of Pharmacopuncture
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    • v.4 no.1
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    • pp.5-21
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    • 2001
  • The influence of electroacupuncture (EA), a traditional Chinese medical treatment, on type Ⅱ collagen-induced arthritis (CIA) was examined in DBA/1J mice in vivo. Mice were immunized intradermally twice at the 3-week interval with bovine type Ⅱ collagen(C Ⅱ). EA stimulation, begun on the 21 simultaneously with the second immunization, was applied at the acupoint equivalent to GV4 three times a week for 3 weeks. The results showed that EA delayed the onset, attenuated the severity of arthritis, and reduced the anti-collagen antibody level. Furthermore, we investigated the impact of EA on the productions of endogenous $interleukin-1{\Beta}$ (IL-1 beta) and prostaglandin E2 (PGE2), and the levels of IL-1 beta mRNA in splenocytes and synovial tissues from C Ⅱ immunized mice on the 45 and cyclooxygenase-2 (COX-2) mRNA in lipopolysaccharide (LPS)-stimulated macrophages of normal mice by using reverse transcriptase-polymerase chain reaction (RT-PCR). EA stimulation significant inhibited the concentrations of splenic endogenous IL-1 beta and serum PGE2. The expression of IL-1 beta mRNA in spleen cells was obviously down-regulated and that in synovial tissues was modestly affected by EA. COX-2 mRNA was highly expressed in cultured peritoneal macrophages when stimulated with LPS. Previous treatment with EA also reduced LPS-stimulated induction of COX-2 mRNA. These data suggest that EA has an inhibitory effect on murine CIA, and the partial mechanism of its therapeutic result may be attributed to inhibiting the productions of IL-1 beta and PGE2 by suppression the IL-1 beta and COX-2 gene activations.

Immunochemical Studies of Starfish Gangliosides: Production of Monoclonal Antibody against AG-2, the Major Ganglioside of Starfish Acanthaster planci, and Detecting Its Distribution in Tissues by TLC Immunostaining

  • Miyamoto, Tomofumi;Yamamoto, Atsushi;Sakai, Maki;Tanaka, Hiroyuki;Shoyama, Yukihiro;Higuchi, Ryuichi
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.298-304
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    • 2006
  • In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.

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Effects of Schizandra chinensis fructus on the Immunoregulatory Action and Apoptosis of L1210 cells (오미자 면역조절작용 및 L1210 세포의 apoptosis 에 미치는 효과)

  • Kwon, Jin;Lee, Se-Jin;So, June-No;Oh, Chan-Ho
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.384-388
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    • 2001
  • The effects of MeOH extracts of Schizandra chinensis fructus (SZX) on the immunoregulatory effect (lymphocyte proliferation, subpopulation, nitric oxide production, phagocytic activity) and apoptosis $(sub-G_1\;peak)$ of L1210 cells were examined. The proliferation of splenocytes and thymocytes were enhanced by the addition of $10\;{\mu}g/mL$ of SZX. SZX were administered p.o. once a day for 7 days in adult male BALB/c mice. SZX resulted in altering subpopulation of splenic B and/or T and thymic T lymphocytes, especially the number of $T_H$ cells were markedly increased by the treatment of SZX in vivo and in vitro. SZX treatment induced the apoptotic cell death in L1210 mouse leukemia cells. In addition, SZX accelerated the production of nitric oxide and phagocytic activity in peritoneal macrophages. These results suggest that SZX have an immunoregulatory property and anti-cancer action.

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Effects of Lonicerae Caulis (LC) on Allergic Contact Dermatitis (ACD) induced by DNCB in mice (인동등(忍冬藤)이 DNCB로 유발된 알레르기성 접촉피부염에 미치는 영향)

  • Lee, Yu-Jin;Lee, Ho-Chan;Choi, Jung-Hwa;Kim, Jong-Han;Park, Soo-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.30 no.3
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    • pp.46-61
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    • 2017
  • Objectives : Allergic contact dermatitis(ACD) is a type IV delayed hypersensitivity reaction that results from cumulative exposures and subsequent sensitization to an environmental chemical. Lonicerae Caulis(LC) can clear away heat and relieve toxin, disperse wind and heat, dredge the channel. The present study was designed to investigate the effects of LC on allergic contact dermatitis(ACD) induced by 2,4-dinitrochlorobenzene(DNCB) in mice. Methods : In this experiment, the effects of LC on changes in body weights, ear and dorsum skin thicknesses, ear weights, clinical aspects on the dorsum skin, histopathological changes, spleen/body ratio, cytokines were investigated. In addition, the effects on proliferations of splenocytes were also investigated in vitro and vivo study. Results : LC spread(SPR) group and LC spread and administered(SPR+ADM) group showed diminished ear thicknesses. In SPR+ADM group, ear weights were lowered significantly compared to contact dermatitis control(CTL) group. LC treatment diminished erythema, desquamation and keratosis which were induced by repeated painting of DNCB. In histopathological observation, spongiosis and edema were diminished in SPR and SPR+ADM group. In cytokines, SPR+ADM group were increased in IL-10, and SPR and SPR+ADM group were decreased in TNF-${\alpha}$ compared with control group. Conclusions : These data suggest that LC can decrease symptoms of ACD, then LC is useful to treat patient with ACD.

Enhancement of Murine NK cell Activity in vitro by Red Ginseng Acidic Polysaccharide

  • Choi, Hye-Sook;Sohn, Eun-Wha;Rhee, Dong-Kwon;Pyo, Suh-Kneung
    • Journal of Ginseng Research
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    • v.33 no.4
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    • pp.278-282
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    • 2009
  • The in-vitro immunomodulatory function of the activity of murine natural-killer (NK) cells induced by redginseng acidic polysaccharide (RGAP) was examined. RGAP induced the significant enhancement of NK cell activity against the Yac-1 tumor cells. The treatment of splenocytes cultured with RGAP for 16 h resulted in a significant increase in NK activity at the E:T ratio of 100:1, and in a 239 and 250% increase at 10 and $100{\mu}g$/ml, respectively. We also demonstrate that RGAP treatment increased the production of interferon (IFN)-$\gamma$ (17-125%) and tumor necrosis factor (TNF)-${\alpha}$ (15-100%), suggesting that the increase in NK cell cytotoxicity could be due to the enhancement of the NK cell production of both cytokines. In addition, RGAP had a stimulating effect on lymphocyte proliferation in the presence of mitogens. Overall, these results suggest that RGAP has an immunopotentiating effect on NK cells, which can support the development of clinical studies on RGAP.

Diol-ginsenosides from Korean Red Ginseng delay the development of type 1 diabetes in diabetes-prone biobreeding rats

  • Ju, Chung;Jeon, Sang-Min;Jun, Hee-Sook;Moon, Chang-Kiu
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.619-626
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    • 2020
  • Background: The effects of diol-ginsenoside fraction (Diol-GF) and triol-ginsenoside fraction (Triol-GF) from Korean Red Ginseng on the development of type 1 diabetes (T1D) were examined in diabetes-prone biobreeding (DP-BB) rats that spontaneously develop T1D through an autoimmune process. Methods: DP-BB female rats were treated with Diol-GF or Triol-GF daily from the age of 3-4 weeks up to 11-12 weeks (1 mg/g body weight). Results: Diol-GF delayed the onset, and reduced the incidence, of T1D. Islets of Diol-GF-treated DP-BB rats showed significantly lower insulitis and preserved higher plasma and pancreatic insulin levels. Diol-GF failed to change the proportion of lymphocyte subsets such as T cells, natural killer cells, and macrophages in the spleen and blood. Diol-GF had no effect on the ability of DP-BB rat splenocytes to induce diabetes in recipients. Diol-GF and diol-ginsenoside Rb1 significantly decreased tumor necrosis factor α production, whereas diol-ginsenosides Rb1 and Rd decreased interleukin 1β production in RAW264.7 cells. Furthermore, mixed cytokine- and chemical-induced β-cell cytotoxicity was greatly inhibited by Diol-GF and diol-ginsenosides Rc and Rd in RIN5mF cells. However, nitric oxide production in RAW264.7 cells was unaffected by diol-ginsenosides. Conclusion: Diol-GF, but not Triol-GF, significantly delayed the development of insulitis and T1D in DP-BB rats. The antidiabetogenic action of Diol-GF may result from the decrease in cytokine production and increase in β-cell resistance to cytokine/free radical-induced cytotoxicity.

Studies on Immunoregulatory Effects of Bowon-tang in the Immune Cells (保元湯의 免疫調節 作用에 관한 硏究)

  • Hwang, Joo-Min;Jung, Myung;Cho, Jung-Hoon;Lim, Kyu-Sang;Yun, Young-Gab
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.4
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    • pp.92-110
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    • 2015
  • Objectives : The water extract of Bowon-tang composited with thePanax, AstragalusandGlycyrrhiza Radixhas been traditionally used for treatment of a sickly child and smallpox in oriental medicine. However, little is known about the regulatory effects of Bowon-tang on the production, expression and activity of immune mediators [nitric oxide, prostaglandin E2, inducible nitric oxide synthetase, cyclooxygenase-2], the macrophage activation factor production, the proliferation, subset expression, the killing activity, and the capping in immune cells.Methods : In this study, we investigated the effects of water extracts from Bowon-tang,Panax, AstragalusorGRin mouse immune cells or human Jurkat T cells. Each extract (25-200 ㎍/㎖)perse had no cytotoxic effect in unstimulated macrophages, but concentration-dependently regulated NO and PGE2production, iNOS expression, and COX-2 activity in mouse peritoneal macrophages with MAF stimulation. These regulatory effects were synergistically increased by their combination (Bowon-tang).Results : The extract of Bowon-tang concentration-dependently regulated T cell proliferation, CD4+and CD8+expression, and NK killing activity in mouse splenocytes and capping in Jurkat T cells.Conclusions : These results suggest that the water extract of Bowon-tang composited with thePanax, AstragalusandGRmay be useful for therapeutic drugs against a sickly constitution and immune diseases, probably by regulating the production of immune mediators.

Comparative evaluation of the murine immune responses to Salmonella enterica serovars Enteritidis, Gallinarum and Typhimurium infection (Salmonella enterica serovars Enteritidis, Gallinarum 및 Typhimurium의 마우스 면역반응의 비교평가)

  • Kim, Kiju;Kim, Dooree;Sun, Jisun;Park, Soyeon;Cho, Youngjae;Ko, Hyun-Jeong;Joo, Hong-Gu;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.53 no.2
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    • pp.95-101
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    • 2013
  • The study was carried out to evaluate and compare the immune responses in mice experimentally infected with either wild-type or isogenic mutants of Salmonella enterica serovars Enteritidis (SE), Salmonella Typhimurium (ST) and Gallinarum (SG). The mutant strains were constructed by allelic replacement of some virulence-associated genes in the wild-type strains. Seven-week-old female BALB/c mice were orally or intraperitoneally inoculated by injecting bacterial suspension. To evaluate the immune responses, enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot (ELISPOT) assay were conducted with serum and fecal samples. As a result, the mice group infected orally with the SE mutant strain showed the highest level of specific IgA-secreting splenocytes, compared to the other groups. The peritoneally injected groups showed the greater levels of IgG1 than the orally injected groups, which was in a good agreement with the previous studies. In addition, the mutant infected groups had the similar secretion levels of antibodies with the wild-type infected groups. These results demonstrated that the SE mutant strain elicited humoral immune response as much as wild-type, implying that it can be useful as a delivery vehicle as well as a candidate of a live attenuated vaccine.

Anti-allergic effect of Lactobacillus rhamnosus IDCC 3201 isolated from breast milk-fed Korean infant (한국인 모유영양아의 분변에서 분리한 Lactobacillus rhamnosus IDCC 3201의 항 알레르기 효과)

  • Lee, Seung-Hun;Kang, Jae-Hoon;Kang, Dae-Jung
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.18-24
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    • 2016
  • We investigated 23 lactic acid bacteria isolated from Korean breast milk-fed infant in order to select strains which show superior anti-allergic effect. The candidates were cultivated and then we obtained dried powders of tyndallized cells and supernatant concentrate separately. Screening was carried out with down-regulation of interleukin (IL) 4 and up-regulation of IFN-${\gamma}$ in mouse splenocytes. As a result of the screening, we selected Lactobacillus rhamnosus IDCC 3201 (RH3201) for oral feeding to ovalbumin-sensitized BALB/c mice. Oral administration of RH3201 as dead cell bodies and supernatant concentrate suppressed hyper-production of serum immunoglobulin (Ig) E levels compared to vehicle group. Such anti-allergic effects were achieved by improvement of the balance between cytokines produced from type-1 helper T (Th1) and type-2 helper T (Th2) lymphocytes. Therefore, RH3201 has potential to improve atopic symptoms by immunomodulatory effect.