• IMMUNE NETWORK
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• 제13권1호
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• pp.30-33
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• 2013

Previously we showed that biodegradable nanoparticles containing poly-IC or CpG oligodeoxynucleotide (ODN) together with ovalbumin (OVA) were efficient at inducing MHC-restricted presentation of OVA peptides in dendritic cells. The CTL-inducing activities of the nanoparticles were examined in the present study. Nanoparticles containing poly-IC or CpG ODN together with OVA were prepared using biodegradable polymer poly(D,L-lactic acid-co-glycolic acid), and then were opsonized with mouse IgG. The nanoparticles were injected into the tail vein of mice, and 7 days later the OVA-specific CTL activities were measured using an in vivo CTL assay. Immunization of mice with the nanoparticles containing poly-IC or CpG ODN together with OVA elicited potent OVA-specific CTL activity compared to those containing OVA only. In accordance with these results, nanoparticles containing poly-IC or CpG ODN together with OVA exerted potent antitumor activity in mice that were subcutaneously implanted with EG7.OVA tumor cells. These results show that encapsulation of poly-IC or CpG ODN together with antigen in biodegradable nanoparticles is an effective approach for the induction of potent antigen-specific CTL responses in vivo.

• IMMUNE NETWORK
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• 제16권4호
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• pp.233-241
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• 2016

DCs, like the sensory neurons, express vanilloid receptor 1 (VR1). Here we demonstrate that the VR1 agonists, capsaicin (CP) and resiniferatoxin (RTX), enhance antiviral CTL responses by increasing MHC class I-restricted viral antigen presentation in dendritic cells (DCs). Bone marrow-derived DCs (BM-DCs) were infected with a recombinant vaccinia virus (VV) expressing OVA (VV-OVA), and then treated with CP or RTX. Both CP and RTX increased MHC class I-restricted presentation of virus-encoded endogenous OVA in BM-DCs. Oral administration of CP or RTX significantly increased MHC class I-restricted OVA presentation by splenic and lymph node DCs in VV-OVA-infected mice, as assessed by directly measuring OVA peptide SIINFEKL-K^b complexes on the cell surface and by performing functional assays using OVA-specific CD8 T cells. Accordingly, oral administration of CP or RTX elicited potent OVA-specific CTL activity in VV-OVA-infected mice. The results from this study demonstrate that VR1 agonists enhance anti-viral CTL responses, as well as a neuro-immune connection in anti-viral immune responses.

• 대한한의학회지
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• 제32권3호
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• pp.10-17
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• 2011

Objective: To evaluate the immune-stimulatory potential of extracts of Broussonetia kazinoki Siebold (BK) on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Material and Methods: C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. BK (100, 300 or 1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma. Further, the effects of BK on expression of cytokine mRNA in OVA-immunized mice splenocytes were evaluated by RT-PCR analysis. Results: BK significantly enhanced OVA-, LPS-, and Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). BK also significantly enhanced total IgM and OVA-specific IgG1 levels in plasma compared with the OVA control group. Moreover, BK up-regulated significantly the expression of mRNA level of IL-2 and IFN-${\gamma}$ in splenocytes. Conclusions: BK has immune-stimulating activity in an OVA-immunized mouse model system, enhancing the Th1 immune response. BK showed no cytotoxicity in this system, suggesting that BK may be a safe and effective adjuvant in humans.

• Clinical and Experimental Pediatrics
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• 제53권4호
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• pp.481-487
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• 2010

목 적: 최근 여러 연구결과 천식을 포함한 알레르기질환의 발병에 소아 성장시기에 알레르겐이나 내독소(LPS)등과 같은 물질에 노출이 중요한 역할을 하는 것으로 제시되었다. 이에 연구자들은 출생 초기에 기도 점막을 통한 알레르겐과 내독소에의 노출이 성장한 후에 알레르기 기도염증과 과반응성의 발생에 미치는 효과를 생쥐 모델을 통하여 규명하고자 본 연구를 시행하였다. 방 법: 실험동물은 무균 상태의 생후 1주이내의 암컷 BALB/c 생쥐를 사용하였다. 실험동물들에게는 각각 신생생쥐시기에 생리식염수, 1% ovalbumin (OVA), $1.0{\mu}g$ LPS, $1.0{\mu}g$ LPS in 1% OVA를 각 비공을 통하여 투여하였다. 실험동물은 연구 시작 제 35일부터 10일간 5% OVA로 감작시켰으며 최종 기도 감작 10일 후부터 3일간 1% OVA로 기도 항원유발을 시행하였다. 최종 기도유발 48시간 후 체적검사(plethysmography)를 이용하여 비특이적 기도과반응성 검사(Methcholine challenge test)를 시행하였으며 검사 후 실험동물을 희생시켜 검체를 취하여 BAL액내 세포분획, 사이토카인, 혈청 면역글로불린을 측정하였다. 결 과: 1) 메타콜린에 의한 기도과반응성은 생후 초기에 OVA, LPS, OVA/LPS를 투여한 군에서 생리식염수를 투여한 군에 비해 통계적으로 유의하게 억제되었다. 2) OVA, LPS, OVA/LPS를 투여한 군에서 BAL 액내의 호산구수와 IL-4, IL-5가 유의하게 낮았다. 3) 혈청내 OVA 특이 IgE는 OVA, LPS, OVA/LPS 투여군에서 유의하게 감소되었다. 결 론: 본 연구 결과 신생생쥐 시기의 점막을 통한 항원, 내독소의 노출이 성숙한 후 항원에 의한 기도염증 및 과반응성의 발생을 억제하였으며 향후 이러한 효과의 작용기전에 대한 연구가 필요할 것으로 생각한다.

• 한국가축번식학회지
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• 제16권3호
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• pp.199-208
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• 1992

These studies were carried out to investigate optimal physological conditions for in vitro fertilization (IVF) of mouse ova. The unfertilized ova were obtained by superovulation from ICR mice of 4 to 6 weeks old. Tyrode's 280 solution was used as basal media, and pH and osmolality of basal media were adjusted with the supplementation of sodium bicarbonate and sodium chloride, respectively. The optimal pH, and osmolality of culture media and the optimum period of sperm preincubation were examined in fertilization in vitro of mouse ova and the subsequent culture in vitro of embryos. The pH range of media examined was designed from 6.5 to 7.5 with 0.2 interval and the range of osmolality from 250 to 370 mOsm with 20 interval, and the period of sperm preincubation examined was 30, 60, 120, and 180 minutes. The ova developed to 2-cell embryosafter 26hrs. of incubation with preincubated sperm were evaluated as in vitro fertilized ones. The results obtained were summarized as follows: 1. The percentage of in vitro fertilized ova was highest (64.7%) in media of pH 7.1 and lowest (38.0%) in pH 6.7. No significant difference in % fertilized ova was found from the media of pH 7.1 to 7.5. Compared with the result from pH 7.1 medium, the pollyspermy was increased signifciantly (p<0.05) in the media of pH over 7.5 and below 6.9;, and the % degenerated ova was significantly (p<0.05) increased in the media of pH below 6.9. 2. The percentage of in vitro fertilized ova was highest (69.4%) in media of osmolality 330 mOsm and lowest (47.9%) in osmolality 250 mOsm. No significant difference in % fertilized ova was found from the media of osmolality 310 to 350 mOsm. Compared with the result from osmolality 330 mOsm in medium, the polyspermy aws increased significantly(p<0.05) in the media of osmolality over 350 mosmol and blow 290 mOsm, and the % degenerated ova was significantly (P<0.05) increased in the media of osmolality below 290 mOsm. 3. The percentate of in vitro fertlilized ova was highest (62.7%) in media of period sperm preincubation 180 min. and lowest (40.4%) in sperm preincubation 30 minutes. No significant difference in % fertilized ova was found from the media of sperm preincubation 120 to 180 minutes. Compared with the result from sperm preincubation 180 minutes in medium, the polyspermy was low differ no significantly(P<0.05) in the media of period sperm preincubation, and the % degenerated ova was signifciantly(P<0.05) increased in the media of sperm presincubation below 60 minutes.

• Korean Journal of Acupuncture
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• 제22권3호
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• pp.119-135
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• 2005

Objectives : The aim of this study was to investigate the Asthma-suppressive and Immune-regulatory effect of AF-HA(Aristolochiae Fructus Herbal-acupuncture) at Joksamni(St36) in OVA(ovalbumin) induced asthma mouse model. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks(once a week). The mice in the OVA-AF-HA group were treated with AF-HA at St36 for the later 8weeks(3times/week). The mice in the OVA-Needle-prick group were treated with single prick with an injection needle at St36 for the later 8 weeks(3times/week). Results : 1. The lung weight and the total cells in lung of the mice treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. 2. Total leukocytes and eosinophils in BALF of the mice group treated with AF-HA at St36 decreased remarkably compared with those of the OVA-control group. 3. The collagen accumulation in lung of OVA-AF-HA group decreased significantly compared with that of the OVA-control group, 4. The concentrations of IL-4, IL-5, IgE in BALF, and IL-4, IL-5, IL-13 in serum of the mice group treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. 5. The numbers of $Gr-1^+/CD11b^+\;and\;CD11b^+$ cells in lung of the mice group treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. 6. The numbers of $CCR3^+,\;CD4^+,\;CD8^+\;and \;CD3e^+/CD69^+$ cells in lung of the mice group treated with AF-HA at 5136 decreased significantly compared with those of the OVA-control group. 7. The mRNA expressions of $TNF-{\alpha}$, IL-4, IL-5, IL13 in lung of the mice group treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. Conclusion : These results suggest that Aristolochiae Fructus Herbal-acupuncture at Joksamni(St36) may be an effictive therapeutic method to treat asthma.

• 한국식품영양과학회지
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• 제30권5호
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• pp.848-853
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• 2001

이 연구는 난백단백질의 알러지성을 없애기 위해 감마선 조사된 OVA에 대한 가열처리가 항원성(알러지성) 및 구조 변화에 미치는 영향을 조사하기위해 수행되었다. OVA 용액 (2.0 mg/mL)을 1) 가열처리구; 2) 가열 후 조사구; 3) 조사 후 가열처리구로 준비하였다. 시료를 개별적으로 가열처리하였고, 감마선 조사는 10 kGy의 흡수선량을 얻도록 조사하였다. 계란알러지가 있는 환자의 혈청과 OVA에 대해 생산된 단클론항체(M-IgG)와 다클론항체(R-IgG)를 사용하여 확립된 경합간접효소면역분석법을 사용하여 시료용액에 있는 OVA의 항원-항체 반응성을 측정하였다. 열변성된 OVA에 대한 항체반응성은 R-IgG가 6$0^{\circ}C$, M-IgG가 7$0^{\circ}C$부터, 그리고 P-IgE의 경우 8$0^{\circ}C$부터 변화하기 시작하였다. P-IgE는 8$0^{\circ}C$이상의 온도에서 가열된 OVA를 잘 인식하지 못하였다. 반면에, M-IgG와 R-IgGdmlruddn 가열처리된 OVA에 대한 P-IgE의 항원-항체 반응성에 미치는 영향은 처리 순서와는 관계없이 모든 가열온도에서 가열처리에 의한 큰 변화를 나타내지 않았다. 가열처리와 감마선 조사에서 시료용액의 혼탁도가 모두 증가하였고, 가열처리보다는 감마선 조사에 의한 혼탁도의 증가가 매우 높았다. 이 결과들은 가열처리가 감마선 조사되어 감소되어 있는 OVA의 알러지성에 큰 영향을 미치지 않는 것을 나타내었다.

• 한국식품과학회지
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• 제43권1호
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• pp.72-76
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• 2011

The adjuvant effects of Korean mistletoe lectin-C (KML-C) were investigated following the oral administration of KML-C with ovalbumin (OVA) as an antigen. Mice were orally immunized with OVA alone or admixed with various doses of KML-C or cholera toxin (CT), and the titer of OVA-specific antibody in the serum and mucosal secretions were determined. OVA+KML-C-treated mice showed high titers of IgA specific to CT in mucosal secretions. The antibody titers in the serum of OVA+KML-C-treated mice were comparable to those in the serum of OVA+CT-treated mice. When mice were immunized with OVA+KML-C or with CT alone and subsequently injected with OVA on the footpads after the primary immunization, they showed a more significant increase in delayed-type hypersensitivity reactions than when they were administered CT alone. These results suggest that KML-C is a potent immunoadjuvant that enhances both humoral and cellular immunity by the mucosal immune system.

• 한국식품과학회지
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• 제44권6호
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• pp.759-762
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• 2012

이번 실험을 통하여 PEIC가 OVA에 의해 유도된 NF-${\kappa}B$ 활성과 iNOS, COX-2 발현에 어떠한 영향을 미치는지 알아 보았다. PEIC는 OVA에 의해 유도된 NF-${\kappa}B$ 활성을 억제시켰다. 또한 PEIC는 OVA에 의해 유도된 iNOS의 발현도 억제시켰다. 하지만 PEIC는 OVA에 의해 유도된 COX-2 발현은 억제시키지 못하였다. 이러한 결과는 iNOS와 COX-2가 서로 다른 메커니즘에 의해 조절된다는 것을 암시한다. 또한 PEIC는 알러지와 같은 만성적인 질병들을 조절할 수 있는 치료제 개발 및 백신 제조에 중요한 역할을 할 것으로 기대한다.

• 한국식품영양과학회지
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• 제33권7호
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• pp.1169-1174
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• 2004

본 연구에서는 감마선 및 전자선종이 ovalbumin(OVA)의 구조 변화에 미치는 영향을 비교 평가하였다. 표준항원 OVA을 3,5,7 및 10 kGy의 흡수선량으로 감마선 및 전자선 조사하였으며 그에 따른 구조적인 변화는 SDS-PAGE, GPC-HPLC 및 단클론 항체를 사용한 Ci-ELISA법으로 측정하였다. Native OVA분자는 감마선 및 전자선 조사에 의해 소편화 및 응집화 되었으며, 조사 선량이 증가할수록 OVA의 분자량이 감소하였다. 또한 OVA의 면역화학적 인 구조는 방사선 조사에 의해 항원-항체간의 결합능이 감소하는 것으로 나타났으나 감마선 및 전자선종에 따른 차이는 없었다. 이상의 결과들은 식품 알레르겐의 제거 및 면역원의 개발에 대한 전자선 조사의 기초자료로 이용될 수 있을 것으로 사료된다.