• Title/Summary/Keyword: in vitro mouse splenocytes

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Immunomodulatory Potential of Weissella cibaria in Aged C57BL/6J Mice

  • Park, Ho-Eun;Kang, Kyung-Won;Kim, Bum-Seok;Lee, Sang-Myeong;Lee, Wan-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2094-2103
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    • 2017
  • Aging is associated with distinct changes in immune cells and a decline in immune function, leading to increased susceptibility to infection and reduced responses to vaccination. Certain strains of lactic acid bacteria exert beneficial effects on the immune system. Previously, we reported that Weissella cibaria JW15 isolated from kimchi possesses immune stimulatory activity in vitro. In the present study, we further investigated whether oral administration of JW15 improves immune function in aged mice. Eighteen-month-old female mice were administered JW15 daily at low (JW15-L; $1{\times}10^8CFU/mouse$) or high dosage (JW15-H; $1{\times}10^9CFU/mouse$), or with Lactobacillus rhamnosus GG (LGG) using oral gavage. Two-month-old female mice were included as healthy young mice. After 4 weeks, the mice were euthanized and immune profiles were analyzed using whole blood and spleen. In complete blood count analysis, the numbers of white and red blood cells were significantly increased in the JW15-L group compared with those in the old mouse (OM) control group. In addition, administration of either JW15 of LGG resulted in higher numbers of splenocytes in comparison with the OM group. Furthermore, proliferative potentials were higher in all probiotic groups than OM. Cytokines such as IFN-${\gamma}$ and IL-6 were secreted at higher levels in splenocytes isolated from JW15-fed mice than in OM control mice. Similarly, mRNA expression of various cytokines was altered in the JW15 groups. Collectively, these results suggest that JW15 supplementation induces immunomodulatory effects in aged mice and indicate JW15 as a potential probiotic strain to improve immune function in aged animals.

Immunoadjuvanticity of Novel CpG ODN (Oligodeoxynucleotide)

  • Park, Su-Jung;Cho, Hyeon-Cheol;Bae, Keum-Seok;Kim, Soo-Ki
    • Molecular & Cellular Toxicology
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    • v.3 no.1
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    • pp.46-52
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    • 2007
  • In the course of novel TLR (Toll like receptor) 9 ligand, we found novel CpG ODN (Oligodeoxynucleotide) was active in augmenting antibody in mice. However, immune mechanism of new CpG ODNs is unclear. To clarify this, we examined immunoadjuvanticity by employing in vitro and in vivo immune profiles. In brief, in vitro treatment of novel CpG ODN upregulated the expression of TNF-$\alpha$, IL-6, and IL-12 mRNA in macrophages as well as that of IFN-$gamma$ mPNA in mouse splenocytes. In parallel, in vivo injection of novel CpG ODN directly activates macrophages and splenocytess, consequently upregulating MHC class II and CD86. Finally, we demonstrated anti-HBs antibody augmentation of novel CpG ODN. Collectively, this data indicates that novel CpG ODN is immunoadjuvant armed with Th1 typed immune machinery.

The Effects of Taklysodoksan Extract on the Anti-cancer and Immunity (抗癌 및 免疫에 대한 托裡消毒散의 效果;萬病回春方을 중심으로)

  • Kim, Hong-Jin;Choi, Jung-hwa
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.13 no.1
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    • pp.100-115
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    • 2000
  • Taklysodoksan(TSS) was a drug used in the treatment of carbuncle and cellulitis in oriental medicine. The purpose of this Study is to investigate the anti-cancer effect of TSS, the proliferation of immunocytes and nitric oxide(NO) production from peritoneal macrophages of mice. This Study estimated the proliferation of Ll210 cell lines, mouse thymocytes and splenocytes and NO production from peritoneal macrophages in vitro and vivo. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). From the in vitro and vivo Study of TSS treatment, it did not effect the proliferation of L1210 cells. It also did not have any positive prodution of NO in peritoneal macrophages. This results suggest that TSS treatment in WanBingHuiChun(萬病回春) did not have significant anti-cancer effect and immuno-action comparing with TSS treatment of WaiKeZhengZhong(外科正宗).

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Effect of Gleditsiae Spina on Hep G2 cells cytotoxicity and Apoptosis and No (조각자(皂角刺)의 간암세포주(Hep G2)에 대한 세포독성, Apoptosis 및 NO에 대한 실험)

  • Kang, Sung-Youg;Cho, Kyoung-Wha;Han, Jong-Hyun;Cho, Nam-Geun
    • The Journal of Internal Korean Medicine
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    • v.18 no.1
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    • pp.48-61
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    • 1997
  • In this study, antineoplastic activity against human hepatocellular carcinoma cell line(Hep G2) was tested in Gleditsiae Spina. Gleditsiae Spina was extracted with water, and the cytotoxic activity was tested using a calorimetric tetrazolium assay(MTT assay), the apoptosis was tested using a DNA electrophoresis and flow cytometry. The nitric oxide production from mouse peritoneal macrophage was tested using a Griess method. Gleditsiae Spina extracts against the proliferation of Hep G2 cells not showed cytotoxicity at the concentration of less than $100{\mu}g/ml$, and Gleditsiae Spina extracts not showed the cytotoxicity of mitomycin C and the cytotoxicity of cisplatin on Hep G2 cells. Gleditsiae Spina extracts aginist the proliperation of BALB/c 3T3 cells not showed cytotoxicity, the proliperation of mouse thymocytes and splenocytes not showed cytotoxicity at the concentration of less than $100{\mu}g/ml$. Gleditsiae Spina extracts not showed nitric oxide production from mouse peritoneal macrophage in vitro. Gleditsiae Spina was administered orally for 7 days at 300mg/kg increased nitric oxide production from mouse peritoneal macrophage.

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Anthraquinones with Immunostimulating Activity from Cassia tora L.

  • Chung, Ha-Sook
    • Preventive Nutrition and Food Science
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    • v.10 no.3
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    • pp.267-271
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    • 2005
  • Many of plants had been reported having immunostimulating activity. This study reports the immunostimulating activity of Cassia tora L. (Leguminosae) seed, by means of solvent extraction method. Ethanol extract and solvent fractions, n-hexane, chloroform, ethylacetate, n-butanol and aqueous layer of Cassia tora L. seed were tested for immunostimulating activity in vitro. The ethylacetate-soluble fraction caused significant inhibition on the production of nitric oxide by murine macrophages (RAW 264.7), and mouse splenocytes were also stimulated at the concentration of 10 pgfmL. Three anthraquinones, chrysophanol (1), isochrysophanol (2) and aloe-emodin (3) with immunostimulating activity were isolated from the ethylacetate-soluble fraction of Cassia tora L. seed through activity-monitored fractionation and isolation method. These results permit Cassia tora L. to be useful as one the of natural immunostimulating crops.

Ahnak-knockout mice show susceptibility to Bartonella henselae infection because of CD4+ T cell inactivation and decreased cytokine secretion

  • Choi, Eun Wha;Lee, Hee Woo;Lee, Jun Sik;Kim, Il Yong;Shin, Jae Hoon;Seong, Je Kyung
    • BMB Reports
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    • v.52 no.4
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    • pp.289-294
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    • 2019
  • The present study evaluated the role of AHNAK in Bartonella henselae infection. Mice were intraperitoneally inoculated with $2{\times}10^8$ colony-forming units of B. henselae Houston-1 on day 0 and subsequently on day 10. Blood and tissue samples of the mice were collected 8 days after the final B. henselae injection. B. henselae infection in the liver of Ahnak-knockout and wild-type mice was confirmed by performing polymerase chain reaction, with Bartonella adhesion A as a marker. The proportion of B. henselae-infected cells increased in the liver of the Ahnak-knockout mice. Granulomatous lesions, inflammatory cytokine levels, and liver enzyme levels were also higher in the liver of the Ahnak-knockout mice than in the liver of the wild-type mice, indicating that Ahnak deletion accelerated B. henselae infection. The proportion of CD4+interferon-${\gamma}$ ($IFN-{\gamma}^+$) and $CD4^+$ interleukin $(IL)-4^+$ cells was significantly lower in the B. henselae-infected Ahnak-knockout mice than in the B. henselae-infected wild-type mice. In vitro stimulation with B. henselae significantly increased $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected wild-type mice, but did not increase $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected Ahnak-KO mice. In contrast, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, IL-10, RANTES, and tumor necrosis $factor-{\alpha}$ secretion was significantly elevated in the splenocytes obtained from both B. henselae-infected wild-type and Ahnak-knockout mice. These results indicate that Ahnak deletion promotes B. henselae infection. Impaired $IFN-{\gamma}$ and IL-4 secretion in the Ahnak-knockout mice suggests the impairment of Th1 and Th2 immunity in these mice.

Water Extracts of Cultured Mountain Ginseng Stimulate Immune Cells and Inhibit Cancer Cell Proliferation

  • Oh, Chan-Ho;Kang, Pil-Sung;Kim, Jae-Whune;Kwon, Jin;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.15 no.3
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    • pp.369-373
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    • 2006
  • Water extracts obtained from cultured mountain ginseng (CMG) were evaluated for their ability to stimulate immune cells and inhibit cancer cell proliferation. The lymphocyte subpopulation in mouse splenocytes in vivo was significantly increased by the administration of the CMG extract (27.4 mg/mouse). Interleukin-2 and ${\gamma}$-interferon in the mice serum increased up to 30% in CMG extract-treated mice. At a concentration of 1.37 mg/mL, nitric oxide increased up to 400% in the macrophage cell line treated with CMG extract. The CMG extract significantly retarded the proliferation of human acute promyelocytic (HL60), human histiocytic (U937), and mouse lymphocytic (L1210) leukemia cell lines in vitro at concentrations over 2.74-13.7 mg/mL. In addition, CMG extract treatments (1.37 mg/mL and 2.74 mg/mL) lead to the increased expression of the p53 gene and protein in cultured U937 leukemia cell lines. These results indicate that water extracts of CMG are capable of both immune cell stimulation and cancer cell growth inhibition.

Effcts of Dangkiyeumja(當歸飮子) Water Extract of anti-allergic responses and on the Functions of Murine Immunocytes (當歸飮子 水抽出液이 抗ALLERGY 反應과 MOUSE의 免疫細胞機能에 미치는 影響)

  • No, Seok-Seon;Lee, Gi-Nam
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.4 no.1
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    • pp.23-42
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    • 1991
  • This study were done to know the effects of Dangkiyeumja on the in vivo and in vitro immune responses of mice. The recipes of Dangkiyeumja used in this study enhanced such, cellular functions of immunocytes as phagocytic capacity of macrophages, rossett-eforming abilities of splenocytes and metabolic activities of lymphocytes, However, the same recipes decreased the formation of such reactive oxygen intermediates(ROI) as superoxide and hydrogenperoxide from the macrophages. The effects of the same recipes on the in vim immune responses was suppressive on the cellular immune response(CIR)measured by delayed-type hypersensitivity against dinitrofluorobenzene and mildly enhancing for the humoral immune response measured by antibody production against sheep red blood cells. The results of this study could be summarized as follow: 1. Administration of Dangkiyeumja enhanced the phagocytic activity of the murine macrophage. 2. Administration of Dangkiyeumja decreased the formation of ROI in the murine macrophage 3. Administration of Dangkiyeumja increased the number of the splenic rotte forming cells in the mouse. 4. Administration of DangKiyeumja did not effect the antibody production against sheep red blood cells. 5. Administration of Dangkiyeumja depressed the delayed-type hypersenitivity against dinitrofluoro benzene in the mouse. The result of this study suggest that Dangkiyeumja could ameliorate the hypersensitivity reactions by reducing the formation of ROI and decreasing the CIR without affecting the other functions of immunocytes.

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Effect of Kami-Cheongsimyeonjatang on cytokine expression with GATA3 regulation in atopic dermatitis-like skin lesions and IgE hyperproduction induced in NC/Nga mice (IgE 과대생산과 피부염이 유발된 NC/Nga생쥐의 비장세포에서 GATA3 조절에 의한 유전자 발현에 미치는 영향)

  • Park, Seul-Ki;Han, Jae-Kyung;Kim, Yun-Hee
    • Journal of Haehwa Medicine
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    • v.17 no.2
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    • pp.167-183
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    • 2008
  • KCSYJT medicines controlled $CD4^+/IFN-\gamma$, and $CD4^+/CD25^+/foxp3^+$ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates T cells of a NC/Nga mouse same time by anti-CD40/rmIL-4, and interleukin-$1{\beta}$, IL-6, TNF-$\alpha$, and TGF-$\beta$ mRNA outturn that bear in T and B cells decreased remarkably by KCSYJT medicines. Intracellular staining of splenocytes anti-CD40/rmIL-4 plus rmIL-4 stimulated as described in a, assessed after 24 h, KCSYJT exerts a mainly immunosuppressive effect that acts at least partially through suppression of the transcription factor GATA3 expression in $CD4^+$ T cells. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result that Th1 cell and Th2 cell observe to be shifted by cytokine expression with GATA3 regulation by KCSYJT medicines could know that KCSYJT medicines can use usefully in allergy autoimmnune diease.

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Yijung-tang, a Traditional Herbal Formula, Exerts Anti-allergic Effect by Regulating Production of Th2-Type Chemokines and Cytokines (Th2 사이토카인 및 케모카인 분비 조절을 통한 이중탕의 항알러지 효능 연구)

  • Jeong, Soo-Jin;Seo, Chang-Seob;Lee, Mee-Young;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.46 no.2
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    • pp.160-166
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    • 2015
  • Yijung-tang (YJT) is a traditional herbal formula comprising 4 medicinal herbs. In the present study, we performed the simultaneous analysis for three compounds of YJT and examined anti-allergic effects in vitro. The column for separation of three compounds was used Gemini C18 column and maintained at 40$^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems. To evaluate Th2 chemokines, YJT was treated into tumor necrosis factor (TNF)-${\alpha}$ and interferon (IFN)-${\gamma}$-stimulated HaCaT cells, and performed ELISA for thymus and activation regulated chemokine (TARC) and regulated on activation, normal T-cell expressed and secreted (RANTES). To measure Th2 cytokines, YJT was added into primary mouse splenocytes, and performed ELISA for interleukin (IL)-4, 5, 13. Calibration curves were acquired with r2 >0.9999. The contents of liquiritin, glycyrrhizin, and 6-gingerol in YJT were 4.50 mg/g, 11.10 mg/g, and 1.33 mg/g, respectively. YJT inhibited production of TARC and RANTES in TNF-${\alpha}$ and IFN-${\gamma}$-treated HaCaT cells. YJT also reduced production of IL-4, 5, and 13 in primary mouse splenocytes. In conclusion, our data will be a valuable information to improve quality control and anti-allergic effects of YJT.