JY-Pol 접합백신으로 유도된 항페렴구균 항체의 보호효과

이주희,한용문,Lee, Jue-Hee,Han, Yong-Moon 대한약학회 2004 약학회지 Vol.48 No.6

We previously reported that Streptococcus pneumoniae capsule attached to the surface protein (JY-Pol) was protective to systemic pneumococcal infection. The JY -Pol antigen induced IgM, IgG, and IgA in mice and provoked cell-mediated immunity. In this current study, we investigated the effect of anti JY-Pol antiserun and monoclonal antibody C2 (Mab C2) specific for the JY-Pol antigen against the pneumococcal disease. Mice that were given the antiserum survived longer than mice that received antiserum pre-absorbed with S.pneumoniae cells or DPBS as a negative control. Heat-treated anti JY-Pol antiserum resulted in survival rates similar to intact fresh JY-Pol antiserum. Mab C2 isolated from JY-Pol-immunized mice also enhanced resistance of naive mice against the pneumococcal diseaser. This protection by Mab C2 appeared to be mediated by opsonization as determined in a RAW 264.7 monocyte/macrophage cell line. Epitope analysis showed that Mab C2 epitope consisted of glucuronic acid and glucose that blocked the interaction of JY-Pol to the C2. Taken together, these data indicate that the antiserum induced by the JY-Pol, a naturally pneumococcal conjugate formula, mediated the protection by passive transfer, which was confirmed by protective effect of Mab C2.

황련 유래 Antimicrobial Peptide의 Candida albicans 감염 억제효과

이주희(Jue-Hee Lee) 대한약학회 2011 약학회지 Vol.55 No.3

We previously reported the protein isolated from Coptidis Rhizoma (CRP), which has antifungal activity against a fungal pathogen, Candida albicans. In the current study, we investigated what portion in the CRP is responsible for the antifungal activity. For the investigation, the CRP was fractionated on a Shepadex G-50 column. Data resulting from the fractionation, seven fractions were obtained. Fractions (Fr.) I, II, and III eluted initially from the column showed no inhibitory effect on the growth of C. albicans, whereas Fr. IV, V, and VI eluted later revealed inhibition of the growth, and Fr. IV and VI showed potent antifungal activity by broth susceptibility analysis. However, Fr. VI was contained in the CRP more than Fr. IV, which led us to select the VI for the following experiments. In a murine model of a subcutaneous candidiasis caused by C. albicans, the Fr. VI displayed a therapeutic effect on nude mice pretreated with anti-neutrophil monoclonal antibody (RB68C5) and then infected subcutaneously with live C. albicans. At day 16, these mice were healed almost up to 78% of the infected area when compared to infected area of control nude mice that received diluent (Dulbecco's Phosphate-Buffered Saline; DPBS), instead of the Fr. VI (P<0.01). The Fr. VI blocked hyphal formation from blastoconidial form of C. albicans (P<0.01), which might prevent penetration of hyphae to the deeper site of skin and thus helps the healing. In the ionic strength test, the effect of Fr. was influenced by Ca2+ ion just like other known antimicrobial peptides, but the influence was affected at an extremely high concentration such as 500 mM. Thus, such ion-concentration is considered to be meaningless in the clinical situation. Considering all data together, Coptidis Rhizoma is appeared to produce an antimicrobial peptide that has therapeutic effect on subcutaneous infection caused by C. albicans.

Glycycoumarin 감초성분의 항진균효과

이주희(Jue-Hee Lee),이영미(Young Mi Lee),한용문(Yongmoon Han) 대한약학회 2011 약학회지 Vol.55 No.3

Glycycoumarin, a 3-arylcoumarine isolated from Glycyrrhizae radix (a family of Leguminosae), is reported to have anti-bacterial activity. However, its antifungal activity is still unknown. In this present study, the antifungal activity of glycycoumarin (GLM) against Candida albicans, a polymorphic fungus was investigated. Possible mechanism such as blocking of the hyphal induction was also analyzed. By the in-vitro susceptibility analysis, GLM showed anticandidal activity, resulting in an almost complete inhibition of the fungal growth at a concentration of 320 μg/ml, which was equivalent to the efficacy of fluconazole at the same dose. In the murine model of disseminated candidiasis GLM enhanced resistance of mice against the disseminated disease (P<0.05), resulting in 60% protection of GLM-treated mice group during a period of 21-day observation. As for its mechanism of the antifungal activity, GLM blocked hyphal production, one of the important of virulence factors by the fungus, from the yeast form of C. albicans (P<0.01). These data indicate that GLM may contribute to the perspectives that focus on the development of a novel agent with antifungal activity specific for C. albicans infection.

폐렴구균 DNA 백신의 유효성 평가

이주희(Jue Hee Lee),한용문(Youngmoon Han) 대한약학회 2005 약학회지 Vol.49 No.6

Streptococcus pneumoniae is the leading cause of pneumonia and bacterial meningitis. The current polysaccharide vaccine has been reported ineffective in elderly adults and children less than 2 years of age. Thus, in recent many researchers have been focusted on a different approach, DNA vaccine. In our laboratory, we developed a Streptococcus pneumoniae DNA(SPDNA) vaccine. This SPDNA vaccine was formulated by inserting the resion encoding part of the capsule in the S.pneumoniae into the LAMP-1. In present work, with use of the SPDNA vaccine we attmpted to to establish a certain methodology useful for evaluation of effectiveness and immunoresponse of a DNA vaccine. Results showed that the subcutaneous route was the most effective for production of antisera specific for S. pneumoniae in mice. By isotyping analyses, IgM, IgG1, IgG2a, and IgG2b were determined. In addition, INF-γ and IL-4 were predominantly detected. Combination of those data resulted in a pattern of IgG1IL-4, which indicates the immunity towards the Th1 response predominantly. Furthermore, the SPDNA vaccination induced resistance of the CD4+T lymphocyte-depleted mice against disseminated pneumococcal infection. These data appear to be possibly due to activation of CD8+T cell-activation. Taken together, this methodology can be applied for evaluating efficacy and mode of action of a DNA vaccine as minimum criteria.

폐렴구균 DNA 백신의 면역병리학적 측면에서의 안전성 평가

이주희(Jue Hee Lee),한용문(Young Moon Han) 대한약학회 2006 약학회지 Vol.50 No.1

We have previously reported the minimum criteria that can be applied to evaluate efficacy and safety of a DNA vaccine with use of Streptococcus pneumoniae DNA vaccne (SPDNA). The SPDNA was formulated by inserting the DAN sequences that are codons specific for the carbohydrate epitope in the capsule of S. penumoniae by phage display peptide library. Administration of the SPDNA into mice induced both humoral and cell-mediated immunities. The induction was protective even in the absence of CD4+ T lymphocyte in mice. Profiles of cytokine and isotyping of antibody displayed tendency of the Th1. In continuation of these studies, we examined if the efficacy of the SPNDA was provoked by the peptide recognized by codons specific for the capsule. Results showed that the peptide vaccine formulae (SPP) induced protective antibody in mice as did the SPDNA. Involvement of the cell-mediated immunity was also determined. Possible side effects of autoimmune diseases such as myositis and C3a production and tumor-formation were undetectable in mice given 7 times of SPDAN vaccination during entire of 92 days. Even after the frequent immunization, immunogenicity of the SPDNA was observed as determined for antibody production, suggesting that there was no immunotolerance provoked. All together, these examining factors would be applied to measurement of DNA vaccine safety regarding the immunopathological aspect.

황련단백질의 황캔디다 작용기전 및 항피부캔디다증 효과

이주희(Jue Hee Lee),심진기(Jin Kie Shim),한용문(Young Moon Han) 대한약학회 2005 약학회지 Vol.49 No.5

Our previous data showed the protein isolated from Coptidis Rhizoma(CRP) had antifungal activity. In present study, we examined mode of action of the CRP and its activity against subcutaneous candidiasis 엳 새 C. albicans yeast cells. Results showed that the CRP blocked hyphal production from yeast form of C. albicans. The CRP also activated RAW 264.7 monocyte/macrophage cell line, which resulted in nitiric oxide (NO) production from the cells. This activation seemed to increase macrophage phagocytosis to destroy the invaders. Like other antimicrobial peptides, CRP was influenced by ionic strength, thus resulting in a decrease of antifungal activity. In murine model of a subcutaneous candidiasis, the sizes of infected areas of the nude mice given the CRP after subcutaneous injection of C. albicans yeast cells to the dorsal skin were 90% less than those of the nude mice groups that received DPBS instead of the CRP. All data indicate that the CRP, which appeared to act like an antimicrobial peptide and to inhibit the morphological transition from blastoconidia, was effective against the subcutaneous disease.

Ginkgo Terpene의 Candidate albicans로 인한 관절염에 대한 치료효과

이순현,이주희,한용문,Lee, Soon-Hyun,Lee, Jue-Hee,Han, Yong-Moon 대한약학회 2005 약학회지 Vol.49 No.2

Candida albicans, a polymorphic fungus, causes systemic and local infections. Recent reports show that the fungus is a main etiological agent for the arthritis. For trea tment, antifungal drugs and/or rheumatoid drugs are used, but resistance and side effects limit application of the drugs. In search of new sources for treatment of the fungal arthritis, we choose Egb 761 (extract of Ginkgo leaves 761), one of the most popular over-the-counter herbal medicines. The Egb 761 contains two major ingredients such as terpene and flavonoid. In the present study, we examined if the terpene portion of Egb 761 had anti-inflammatory activity against C.albicans-caused arthritis. The terpene was extracted with combination of methanol and water from the Egb 761, followed by gel-permeation chromatography. Presence of terpene was determined by the Salkowski colorimetric method and HPLC analysis. For an animal model of inflammation induction, mice were given an emulsion form of C.albicans cell wall mixed with Complete Freund's Adjuvant (CFA) by footpad-injection. Results showed that intraperitoneal administration of the water-soluble portion that contained terpene and flavonoid reduced the inflammation. Whereas the terpene had anti-inflammatory activity, flavonoid portion had no such activity, For determination of possible mechanism of the activity, the terpene seemed to be suppression of nitric oxide (NO) production from LPS-treated macrophages. Taken together the Ginkgo terpene may have anti-inflammatory effect against C.albicans-caused arthritis, possibly by blocking NO production.

페렴구균 전신감염에 대한 협막. 표면단백질 접합백신의 효과

한용문,이주희,Han , Yong-Moon,Lee , Jue-Hee 대한약학회 2004 약학회지 Vol.48 No.6

ln the present work to determine effect of a Streptococcus pneumoniae conjugate vaccine, S.pneumoniae capsule attached to the surface protein (JY-Pol) was ex amined. This JY-Pol contained approximately 92% and 6% carbohydrate and protein, respectively. Gel electrophoresis revealed the presence of the surface protein in the JY-Pol. By the double immunodiffusion and isotyping ELISA analyses, administration of JY-Pol that was adsorbed to alum adjuvant (JY-Pol/Alum) into mice induced IgM, IgG, and IgA specific for the S.pneumoniae capsule. The ATCC capsular polysaccharide adsorbed to alum (ATCC-Pol/Alum) provoked only IgM in mice. In survival tests, mice that were immunized with the JY-Pol/Alum before intravenous challenge with live S.pneumoniae survived entire period of 46 day-observation, whereas all mice that received ATCC-Pol/Alum or only diluent instead of the vaccination died within 5 and 12 days, respectively. Results from footpad-edema test showed that JY-Pol/Alum formula provoked the cellular immunity as determined by swelling of the mouse footpad. These data indicate that the naturally conjugated JY- Pol enhances resistance of mice against disseminated pneumococcal disease due to S.pneumoniae by both humoral and cellular immune responses.

황련에서 분리된 단백질성분의 항진균효과

김현경(Hyunkyung Kim),이주희(Jue Hee Lee),심진기(Jin Kie Shim),한용문(Young Moon Han) 대한약학회 2005 약학회지 Vol.49 No.4

Antimicrobial peptides are evolutionary ancient weapons for animal and plant species to depend themselves against infectious microbes. In the present study, we investigated if an antimicrobial peptide was produced from Coptidis Rhizoma. For the determination, protein substance from the medicinal plant was isolated by various preparations. Among the preparations, the protein portion dissolved in phosphate-buffered saline solution (CRP-DS) that contained the most amount of protein (90%) resulted in maximal inhibition of Candida albicans which causes local and systemic infections. Analyses by gel-electrophoresis and gel-permeation chromatography showed the CRP-DS formed a single band of approximately 11.8 KDa as molecular size. Antifungal activity of the CRP-DS was almost equivalent to antifungal activity by fluconazole, resulting in MIC (minimal inhibitory concentration) of approximately 50 ㎍/㎖. The antifungal activity was a dose-dependent. The antifungal activity appeared to be inactivated by heat-treatment and ionic strength, respectively. In a murine model, the CRP-DS enhanced resistance of mice against disseminated candidiasis. The HPLC analysis demonstrated maximum 4% of berberine as residual content in the CRP-DS preparation resulted in no influence on the antifungal activity. In addition, protein portion isolated from Phellodendri Cortex producing the alkaloid component like Coptidis Rhizoma had no such anticandidal effect. These results indicate that the protein substance from Coptidis Rhizoma was responsible for the antifungal activity.

대퇴골두 골괴사를 가진 환자에서의 조립식 대퇴주대를 이용한 무시멘트형 고관절 전치환술 -금속 금속 관절면과 세라믹 세라믹 관절면의 비교-

강번중 ( Bun Jung Kang ),이주희 ( Jue Hee Lee ),정영복 ( Young Bok Jung ),김진홍 ( Jin Hong Kim ),한용문 ( Yong Moon Han ),장의찬 ( Eui Chan Jang ),김경운 ( Kyoung Woon Kim ) 대한고관절학회 2011 Hip and Pelvis Vol.23 No.4

목적: 진행된 대퇴골두 골괴사증 환자에서 무시멘트형 조립식 대퇴주대를 이용한 고관절 치환술의 임상적 및 방사선학적 결과를 평가하였으며, 금속-금속 관절면을 사용한 군과 세라믹-세라믹 관절면을 사용한 군 간, 결과에 차이가 있는지를 확인하고자 하였다. 대상 및 방법: 2001년 1월부터 2004년 12월 까지 대퇴골두 골괴사증으로 조립식 대퇴주대를 이용하여 무시멘트형 고관절 치환술을 시행 받았으며, 5년 이상 추시관찰이 가능하였던 66명(78예)을 대상으로 하였고, 술 후 평균 추시 기간은 평균 77개월(60~122개월) 이었다. 결과: 임상적 평가로 술 전 평균 Harris 고관절 점수는 53점에서 최종 추시 시 88.5점으로 향상되었다. 방사선학적 검사 결과에서 대퇴주대의 안정도는 68예에서 골성고정을 얻었으며, 9예에서 섬유성 안정고정, 1예에서 불안정고정을 나타내었다. 재치환술은 3예에서 시행하였으며, 122개월 간 대퇴 스템의 생존율은 95.7%인 것으로 조사되었다. 결론: 진행된 대퇴골두 골괴사증에서 시행한 조립식 대퇴주대를 이용한 무시멘트형 고관절 전치환술은 5년 이상 추시 결과 금속-금속 관절면을 사용한 군과 세라믹-세라믹 관절면을 사용한군, 모두 만족한 만한 임상적 및 방사선학적 결과를 얻었다. Purpose: The purpose of this study was to evaluate the clinical and radiological outcomes after total hip arthroplasty using the S-ROM modular system for osteonecrosis of the femoral head, and to compare the results between the groups using metal-on-metal articulation and ceramic-on-ceramic articulation. Materials and Methods: Sixty-six patients (78 hips) with osteonecrosis of the femoral head were evaluated after primary total hip arthroplasty between January 2001 and December 2004, using an S-ROM proximal modular femoral stem. The average follow-up was 77 months (range, 60 to 122 months) and all patients were followed for more than five years. Results: The average Harris hip score improved from 53 points to 88.5 points at the final follow-up. At the latest radiologic evaluation, sixty-seven stems had bony ingrowth stability, and 10 stems had stable fibrous ingrowth. However, one stem had diffuse extensive osteolysis and loosening, which was revised at 9 years. Postoperative complications included 4 cases of heterotrophic ossificiation, 1 case of linear fracture after insertion of the femoral stem, 1 case of dislocation, 2 cases of infection, and 1 case of extensive osteolysis and loosening. There were 3 cases of revision and Kaplan-Meier survivorship analysis with revision estimated at a 95.7% chance of survival for the femoral component during 122 months. Conclusion: Our study showed that total hip arthroplasty using the S-ROM modular system with metal-on-metal articulation or ceramic-on-ceramic articulation had favorable clinical and radiological mid- to long-term results.