• 대한약침학회지
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• 제19권1호
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• pp.21-27
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• 2016

Objectives: The present study was undertaken to evaluate the immunomodulatory (IM) activity of IM-133N, a herbal combination in various immunotherapeutic experimental models. Methods: The IM activity of IM-133N was evaluated against three experimental models namely, effect of IM-133N against Escherichia coli (E. coli)-induced abdominal sepsis in mice, and carbon clearance test was performed in Wistar albino rats to evaluated the phagocytic potential of IM-133N, in addition IM-133N was evaluated for its immunoglobulin enhancing potential in rats, where the immunoglobulin levels were measured by zinc sulphate turbity (ZST) test. Further, IM-133N was subjected for detailed liquid chromatography-mass spectrometry (LC-MS)/MS analysis to identify the probable active constituents present in it. Results: The findings of the present study has demonstrated very promising IM property of IM-133N in all the experimental models. Briefly, pretreatment with IM-133N at 125, 250, 500 and 1,000 mg/kg, p.o. doses had protected the mice against E. coli-induced abdominal sepsis and mortality, further the effect of IM-133N was found to be significant and dose-dependent. In support of this, in another study administration of IM-133N showed a significant and dose-dependent increase in serum immunoglobulin levels, estimated by ZST test. In line with the above findings, in the carbon clearance test the low doses (125 and 250 mg/kg, p.o.) of IM-133N increased the rate of carbon clearance, whereas the higher doses (500 and 1,000 mg/kg, p.o.) did not sustain the response, and saturation effect was considered as one of the possible reason for futility of higher doses for IM-133N. In addition, A detailed LC-MS/MS analysis of IM-133N showed 17 bioactive phytochemical constituents: namely, apigenin, chaulmoogric acid, mesquitol, quercetin, symphoxanthone, salireposide, ${\beta}$-sitosterol, nonaeicosanol, ${\beta}$-amyrin, betulic acid, oleanolic acid, symplososide, symponoside, symploveroside, symplocomoside, symconoside A and locoracemoside B. Conclusion: These findings suggest that IM-133N possesses significant IM activity and, hence, could be useful for eradicating opportunistic disease-triggering pathogens via immunotherapeutic mechanisms. The findings also suggest IM-133N may also useful in other immunity disorders.

• 대한소아치과학회지
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• 제30권1호
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• pp.61-68
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• 2003

치아우식증을 예방하기 위한 백신연구를 위하여 주원인균인 Sterptococcus mutans 균체항원과 이 균에서 분리한 glucosyltransferase (GTF)를 항원으로 하고 이에 면역조절기능이 있는 retinoic acid (RA)를 첨가하여 투여경로와 백신조성이 이들 항원에 대한 면역반응에 미치는 영향을 실험하였다. 균체항원(Ingbritt strain)을 마우스의 피하에 Complete Freund's Adjuvant와 함께 투여하여 생산되는 혈청내 응집항체가는 serotype e (LM-7)와는 강한 교차반응을 보였으나 serotype f (OMZ-175)와는 거의 교차반응을 일으키지 않았다. 면역혈청내 항-GTF 및 항-Ag I/II 항체중 항-GTF IgA는 피하로 투여시 전혀 검출되지 않았으나 이에 RA를 첨가하면 다량의 항체 생산을 관찰하였고 그 정도는 경구투여시의 생산량을 능가하였다. GTF를 alum과 함께 투여하여 생산되는 혈청내 항-GTF 항체중 IgM은 피하로 투여시 상당량이 검출되었고 RA를 첨가하면 그 생산이 증가되었으며 경구로 투여시 대조군에 비하여 약간증가를 보였으나 피하투여시의 그것에는 미치지 못하였다. GTF-특이 IgG는 경구투여시는 전혀 검출되지 아니하였고, 피하투여시에만 현저한 증가를 보였으며, RA첨가는 이에 영향을 미치지 못하였다. 항-GTF IgA는 피하로 투여시 전혀 검출되지 아니하였으나 이에 RA를 첨가하면 증가된 항체생산을 관찰하였고 그 정도는 경구투여시의 생산량을 능가하였다. 이상의 실험성적은 GTF에 대한 항체생산은 투여경로와 항원의 종류에 따라 다양한 반응을 나타내며 RA는 이를 백신에 첨가하면 피하경로를 이용하여 면역하더라도 경구투여와 유사한 IgA-매개 면역반응으로 조절시킬 수 있는 가능성을 나타내었다.

• 대한바이러스학회지
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• 제28권1호
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• pp.71-78
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• 1998

Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

• 생명과학회지
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• 제30권10호
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• pp.896-904
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• 2020

본 연구에서는 가축의 면역증강용 생균제 개발을 위하여 Pediococcus acidilactici SRCM102607의 프로바이오틱스 특성 및 면역활성을 조사하였다. 전통발효식품으로부터 유산균을 분리 하였고, 분리 유산균을 대상으로 가축유해 미생물 5종에 대한 항균활성을 측정하였다. 우수한 결과를 나타내는 5종의 유산균을 1차 선별하였고, 이를 대상으로 생균제 소재 활용 가능성을 확인하기 위해 용혈성, 담즙산염 분해효소, 항산화 활성 분석을 실시하여 최종적으로 SRCM102607을 선별하였으며, 16S rRNA 유전자 염기서열 분석을 통해 Pediococcus acidilactici SRCM102607로 명명하였다. SRCM102607의 pH 2 조건에서 내산성은 1.54×10⁵ CFU/ml의 생균수를 보였으며, 0.5% 이상의 oxgall이 포함된 조건에서도 10⁵ CFU/ml 이상의 높은 생균수를 나타내었다. 또한, 선발균주의 산업적으로 활용할 수 있는 가능성을 검토하기 위해 항생제 내성 및 분해 효소능을 측정하였고 다양한 항생제에 대한 내성과 유해 효소를 생성하지 않음을 확인하였고, 최종적으로 면역 증강제로서의 활용 여부를 확인하기 위해 TNF-α 생성능(171.86±4.00 ng/ml)을 확인하였다. 본 연구를 기반으로 SRCM102607은 가축 생균제 소재로 활용 가능성과 면역활성이 뛰어난 유산균으로 생균제 산업에서의 잠재적 적용 가능성을 확인하였다.

• 생약학회지
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• 제36권2호통권141호
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• pp.129-135
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• 2005

Lectins from Allomyrina dichotoma (ADL) and Bombyx mori (BML) were partially purified by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. An assay for cytokine expression was carried out by using reverse transcription polymerase chain reaction(RT-PCR). mRNA isolated from PBMC(human peripheral blood mononuclear cells) were stimulated with ADL(O.D.=0.2) and BML(O.D.=0.1) for various times(1,4,8,24,48 and 72 h) and various cytokine mRNA assessed by RT-PCR were shown as follows: The patterns of bands for IL-1 mRNA of BML were very similar with those from ADL and these bands were decreased along the increasing reaction times after showing a strong band at 1 h. However mRNA expressions for IL-2, IL-6, $IFN{\gamma}$ and $TNF{\alpha}$ showed different patterns between ADL and BML. With the effect of ADL, the expression of IL-2 and IL-6 mRNA were continuously detected until 72 h with the strongest band of IL-2 mRNA at 24 h. The strong bands of $IFN{\gamma}$ mRNA were observed from 4 to 8 h but the strongest one of $TNF{\alpha}$ was just observed at 1 h. Meanwhile with BML, the bands for IL-2 and $IFN{\gamma}$ were increased along the increasing reaction times until 72 h. The strongest bands were showed from 4 to 8 h with IL-6 and at 8 h with $TNF[\alpha}$. To verify quantitatively ELISA was used for assay of protein secretions of the cytokine gene with IL-2 and $IFN{\gamma}$ expressed markedly different in RT-PCR. The highest cytokine secretion for IL-2 was demonstrated at 48 h. The production of $IFN{\gamma}$ was markedly increased at 24 h and secreted highest at 72 h. These result suggest that ADL and BML, as inducers of cytokines, can elicit detectable cytokine mRNA from PBMC within the first few hours of stimulation and maintain the production of cytokines for a few days by the methods of RT-PCR and ELISA.

• 한국식품과학회지
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• 제45권3호
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• pp.385-390
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• 2013

본 연구에서는 참치 추출물 및 분획물들에 의한 NO 생성에 미치는 영향을 살펴보았고 참치 85% aq. MeOH 분획물을 중심으로 면역과정의 생물학적 작용과 대사적 변화를 유도하는 IL-6, IFN-${\gamma}$ 및 TNF-${\alpha}$ 같은 사이토카인의 생성을 측정하여 참치 추출물에 의한 면역 기능 조절에 대하여 검토하였다. 참치 추출물과 각 분획물들은 control보다 낮은 NO 생성량을 나타내었으며, 특히 85% aq. MeOH 및 n-hexane 분획물에 의한 저해효과가 높았다. IL-6와 TNF-${\alpha}$의 경우, 참치 85% aq. MeOH 분획물을 농도별(1, 3 및 $10{\mu}g$)로 처리했을 때 그 중 $10{\mu}g/mL$ 첨가농도에서 6시간 배양 후 가장 낮은 IL-6 및 TNF-${\alpha}$ 생성량을 확인할 수가 있었다(p<0.05). IFN-${\gamma}$의 생성은 참치 85% aq. MeOH 분획물(1, $3{\mu}g/mLg$ 첨가농도)을 LPS와 함께 처리 했을 때 24시간과 48시간 배양 시 생성량이 증가하는 경향이었으나 유의적 차이는 없었다. 따라서 참치 85% aq. MeOH 분획물은 NO 생성과 pro-inflammatory 사이토카인(IL-6와 TNF-${\alpha}$)을 감소시켜 염증반응을 예방할 것으로 기대된다.

• 한국미생물·생명공학회지
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• 제35권2호
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• pp.118-127
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• 2007

본 연구는 마우스를 대상으로 유기게르마늄 강화효모 경구투여에 의한 면역조절작용 효과를 확인하고자 하였다. 마우스를 대상으로 9일간 경구투여한 결과 대조군인 게르마늄 비강화 효모 투여군에 비해 복강대식세포, B세포, NK 세포의 활성이 현저히 증가한 것으로 확인되었으며, 최종 실험결과 대식세포는 게르마늄 강화효모 투여 후 식세포활성, 주화성, 부착성, rosette 형성능 현저히 증가하였다. Superoxide $anion(O_2^-)$ 생성능은 대조군에 비해 유기게르마늄 강화군 투여군에서 3배 활성이 증가하였으며, NO 생성능과 $TNF-{\alpha}$ 생성능도 농도의존적으로 증가하였다. B-세포 활성화에 의한 cytolytic activity 증가에 의한 PFC형성능도 게르마늄 비강화 효모에 비해 현저히 증가하였으며 상업화 유기게르마늄으로 알려지고 있는 Ge-132에 비해 2배 이상 높은 활성이 확인되었다. Cytotoxic acivity에 의한 항 종양활성에서는 양성대조군인 Doxorubicin 투여군에서와 유사한 저해활성을 나타내었으며 고용량 유기게르마늄 효모(2,400 mg/kg) 투여시 60%의 종양활성 억제효과가 확인되었다. 이러한 결과를 종합해 볼 때 유기게르마늄 강화효모가 실험동물 뿐만 아니라 인체의 유용한 면역조절제로서의 이용성이 기대된다.

• 생약학회지
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• 제34권3호통권134호
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• pp.210-217
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• 2003

Korean mistletoe (Viscum album) extract has been found to posses immunostimulatory activity. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract might activate mouse splenic macrophages to produce tumor necrosis $factor-{\alpha}\;(TNF-{\alpha}$) and might play a role in anticancer. To know the effect of M11C on the production of $TNF-{\alpha}$, the splenic macrophages were treated by the M11C, and then collected the supernatant (M11C stimulated splenic macrophage-conditioned media; MSCM). MSCM was analyzed for the $TNF-{\alpha}$ secretion by means of ELISA and immunoblotting, and mRNA expression was analyzed by RT-PCR. The S-180 murine sarcoma model was established to know the effect of M11C on the inhibition of tumor growth. M11C had the effect of $TNF-{\alpha}$ production from splenic macrophages performed by ELISA technique. This ELISA data was reconfirmed by immunoblotting assay. The effects of M11C on the expression of $TNF-{\alpha}$ mRNA from the macrophages was also shown. M11C also had the inhibitory effect of S-180 tumor growth. These data suggest that Korean mistletoe extract M11C may be used for an immunomodulator.

• 대한예방한의학회지
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• 제5권1호
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• pp.103-115
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• 2001

Background : Gamisamryungbaekchul-san(加味蔘?白朮散) is a herbal medicine which has been used for the traditional therapeutic agent of augmentation of the spleen and reinforcement of the Qi. Objective : This Study was performed to investigate the effect of Gamisamryungbaekchul-san on the tumor and immune response in the moose B16 melanoma tumor model. Materials and Methods : The tumor was induced by subcutaneous inoculation of B16BL6 melanoma cells in the shaved dorsal region of mice. Mice were orally administered with Gamisamryungbaekchul-san extract(26.3mg/mouse) for 14days after inoculation. For making examination of antitumor effect, the Increase of life span, Tumor growth inhibition rate, change of body weight were measured and evaluated. For the immune response increasing effect, the percentage of T lymphocyte and B Lymphocyte in the peripheral blood, the percentage of CD4+ T-cell, CD8+ T-cell and CD4+/CD8+ ratio in the peripheral blood and spleen, interleukin-2 productivity were measured and evaluated. Results : Gamisamryungbaekchul-san showed 16.59% increase of life span, 31.64% tumor growth inhibition rate and increase of body weight. Gamisamryungbaekchul-san increased the percentage of T lymphocyte in the peripheral blood, CD4+ T cell percentage of peripheral blood and spleen, and Interleukin-2 productivity as compared with the Control group. Whereas Gamisamryungbaekchul-san had no effect on the percentage of B lymphocyte in the peripheral blood, the percentage of CD8+ T cell, CD4+/CD8+ T-cell ratio in both of peripheral blood and spleen as compared with the Control group. Conclusion : This study shows that Gamisamryungbaekchul-san has anti-tumor effects and immunoregulatory effects on the B16 melanoma tumor model. It is suggested that Gamisarmyungbaekchul-san could be a useful immunomodulator and anti-tumor agent.

• 생약학회지
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• 제24권4호
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• pp.267-281
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• 1993

To find antitumor components from higher fungi, the mycelia of Collybia confluens (Pers. ex Fr.) Kummer were cultured in artificial media. For efficient production of the mycelia, the influences of various modifications of culture conditions were examined. A water-soluble protein-bound polysaccharide fraction, Fr. A, was obtained from the mycelia by hot water extraction. When Fr. A was purified and fractionated by DEAE-cellulose and Sepbadex G-200 gel filtration chromatographies into four fractions which were designated B, C, C-I and C-II. The tumor inhibition ratios of these fractions ranged from 46% to 75% against the solid forms of sarcoma 180 in ICR mice at doses of 20 and 50 mg/kg/day when given intraperitoneally. Especially, Fr. C which was named Collyban(CB) exhibited a marked life-prolonging effect of the mice against ascitic forms of sarcoma 180 at a dose of 50 mg via i.p. administration. To extend spectra of the antitumor activities and eliminate the effects of allograft rejection, the characterization of antitumor effects of CB was performed in syngeneic host-tumor systems. It did not show any antitumor activity against L1210 murine leukemia in $CD_2Fl$ mice but prolonged their life span against ascitic forms of $MM_{46}$ carcinoma in $C_3H/He$ mice. Also it exhibited antitumor activity against human cervical cancer HeLa cells that were xenografted into nude mice having BALB/c genetic backgrounds by the i.p. injection at a dose of 100 mg/kg/day. In order to characterize the antitumor components, CB was examined by chemical analysis. It was acidic protein-bound polysaccharides composed of 31% polysaccharide, 27% protein and 3% hexosamine. CB was fractionated into two fractions, Fr. C-I(M.W.: 500 Kd) and Fr. C-II(M.W.:30 and 8 Kd) by Sephadex G-200 gel filtration chromatography.