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녹동균 세포외막 단백질 백신 CFC-1-101의 안정성 및 면역원성 검토 : 임상 제 Ⅰ/Ⅱa상 시험
장인진,김익상,유경상,임동석,김형기,신상구,장우현,박완제,이나경,정상보,안동호,조양제,안보영,이윤하,김영지,남성우,김현수 대한감염학회 1998 감염 Vol.30 No.3
목적 : 제일제당에서는 녹농균의 세포외막 단밸질을 유효성분으로 하는 백신인 CFC-101을 개발하였으며, 동물시험에서 이 백신의 안전성과 유효성을 입증하였다. 본 연구에서는 이 녹농균 백신의 인체에 대한 안전성과 면역원성을 평가하는 동시에 인체 접종시의 최적 투여 용량을 결정하기 위하여 제 I/Ⅱa상 임상시험을 수행하였다. 방법 : 건강한 성인 남자를 피험자로 선별하여 각 용량군에 백신투여자 6명, 위약투여자 2명을 배정하였다. 백신 투여군은 0.25mg, 0.5mg 또는 1.0mg 용량의 녹농균 백신을 7일 간격으로 3회에 걸쳐 근육주사 하였으며, 위약 투여군에게는 세포외막 단백질을 제외한 동일한 성분을 투여하였다. 백신접종 후 국소적 또는 전신적인 반응의 발생여부를 관찰하고, 혈액시료를 체취하여 백신의 역가와 유효성을 검정하였다. 결과 : 녹농균 백신 CFC-101은 모든 접종자에서 양호한 내약성을 보였다. 또한 0.5mg 과 1.0mg 백신 투여군에서는 100%의 항체양전율을 나타내었다. 생성된 항체는 녹농균 세포외막단백질에 특이성을 보였고, 녹농균 감염에 대해 방어효능이 있었다. 결론 : 이와같은 결과로부터 이 녹농균 백신은 인체에 안전하게 투여할 수 있으며, 높은 항체 생성능으로 감염방어 효능을 보이고 0.5mg과 1.0mg이 최적용량인 것으로 판단되었다. Background : We developed a Pseudomonas aeruginosa outer membrane protein(OMP) vaccine CFC-101, and the prophylactic efficacy of which has been demonstrated in animal models. In order to evaluate the safety and immunogenicity of the P. aeruginosa vaccine, we carried out a phase I/Ⅱa clinical trial in healthy male volunteers. Methods : Groups of eight volunteers, including two placebo subjects, were vaccinated intramuscularly with three doses of 0.25, 0.5 or 1.0 mg of the vaccine at one week intervals. Sings of systemic and local reactions observed after vaccination were recorded for each vaccinee for 5 days. Physical examinations were performed on days 0, 1, 7, 8, 14, 15, 21, and 42, and clinical laboratory tests were done on days 0, 3, and 21. Blood samples for assay of serum antibody levels were obtained up to 42 days after the first vaccination. Results : The vaccine was generally well tolerated by all vaccinees, showing no significant side effects. In the three dosage groups, all vaccinees, except one receiving the 0.25 mg dose, showed significant elevation in serum IgG antibody titers against the vaccine proteins, indicating 100% seroconversion in 0.5 and 1.0 mg groups. The human antibodies induced by the vaccine were specific for P. aeruginosa OMPs, as confirmed by western blot analysis and immunoprecipitation assays. The capacity of the human antisera to enhance opsonophagocytic killing activity by polymorphonuclear leukocytes and to confer protection against P. aeruginosa infections indicates that the antibodies elicited by the vaccine have protective efficacy. Conclusion : We conclude that the P. aeruginosa OMP vaccine is safe and effective for human use and its optimal dose to be 0.5 or 1.0 mg.
LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 양파(Allium cepa L.) 껍질 에탄올 추출물의 항염증 효과
안나경(Na-Kyung Ahn),강보경(Bo-Kyeong Kang),김꽃봉우리(Koth-Bong-Woo-Ri Kim),김민지(Min-Ji Kim),배난영(Nan-Young Bae),박지혜(Ji-Hye Park),박선희(Sun-Hee Park),안동현(Dong-Hyun Ahn) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.11
양파 껍질 에탄올 추출물(OPEE)의 in vitro 및 in vivo에서 항염증 효과를 알아보기 위해 lipopolysaccharide(LPS)로 활성화된 RAW 264.7 세포로부터 분비되는 염증 관련 매개인자들의 발현량과 ICR 마우스모델을 이용한 귀 부종 및 조직 분석을 진행하였다. NO 및 염증성 cytokine(IL-6, IL-1β 및 TNF-α)의 분비량이 OPEE에 의해 농도 의존적으로 감소함을 확인하였다. iNOS 및 COX-2의 발현량은 LPS에 의해 증가하였으나, OPEE 처리에 의해 농도 의존적으로 발현량이 감소하였다. 전사인자인 NF-κB의 활성과 인산화효소인 MAPKs(p-38, JNK 및 ERK)에서도 LPS에 의해 증가된 발현량이 OPEE에 의해 억제됨을 확인하였다. 따라서 NO 및 염증성 cytokine의 분비량 감소는 NF-κB와 MAPKs의 활성 저해에 의한 것임을 확인하였다. In vivo 실험에서는 croton oil로 귀 부종을 유발한 마우스에 OPEE를 처리하였을 때 prednisolone 처리구와 유사한 수준으로 귀 부종이 억제되었으며, 귀 조직 관찰에서도 경피 및 진피의 두께와 진피로 침윤된 mast cell의 수도 억제됨을 확인하였다. 이상의 결과 OPEE는 항염증 소재로서 염증을 예방하거나 치료하는 데 이용 가능성이 높음을 제시한다. Inflammation is a complex process involving a variety of immune cells, which defend the body from harmful stimuli. However, pro-inflammatory cytokines and inflammatory mediators can also exacerbate diseases such as cancer. Onion peel contains several phenolic compounds, including quercetin at an amount 20 times greater in peel than edible flesh. Therefore, in this study, the anti-inflammatory effects of onion peel ethanol extract (OPEE) were investigated lipopolysaccharide-induced inflammatory response. In our results, NO production decreased in a dose-dependent manner. Secretion of IL-6, TNF-α, and IL-1β was suppressed by 44%, 53%, and 60% respectively, at 100 μg/mL. Moreover, OPEE also suppressed expression of COX-2, iNOS, NF-κB, and MAPKs in a dose-dependent manner. Formation of mice ear edema was reduced at the highest dose tested compared to the control, and reduction of ear thickness was observed in the histological analysis as well. In the acute toxicity test, no morality was observed in mice administered 5,000 mg/kg body weight of OPEE over a 2-week observation period. These results suggest that OPEE may have significant effects on inflammatory factors and be a potential anti-inflammatory material.
( Na Young Kim ),( Kyung-eui Park ),( Yong Jin Lee ),( Yeong Mun Kim ),( Sung Hyun Hong ),( Won Rak Son ),( Sungyoul Hong ),( Saehyung Lee ),( Hye Bin Ahn ),( Jaehyuk Yang ),( Jong-pil Seo ),( Yoon-Ky 한국미생물생명공학회 2019 Journal of microbiology and biotechnology Vol.29 No.7
Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are the most toxic substances known. However, the number of currently approved medical countermeasures for these toxins is very limited. Therefore, studies on therapeutic antitoxins are essential to prepare for toxin-related emergencies. Currently, more than 10,000 Halla horses, a crossbreed between the native Jeju and Thoroughbred horses, are being raised in Jeju Island of Korea. They can be used for equine antitoxin experiments and production of hyperimmune serum against BoNT/A1. Instead of the inactivated BoNT/A1 toxoid, Halla horse was immunized with the receptor-binding domain present in the C-terminus of heavy chain of BoNT/A1 (BoNT/A1-HCR) expressed in Escherichia coli. The anti-BoNT/A1-HCR antibody titer increased rapidly by week 4, and this level was maintained for several weeks after boosting immunization. Notably, 20 μl of the week-24 BoNT/A1-HCR(-immunized) equine serum showed an in vitro neutralizing activity of over 8 international units (IU) of a reference equine antitoxin. Furthermore, 20 μl of equine serum and 100 μg of purified equine F(ab’)<sub>2</sub> showed 100% neutralization of 10,000 LD<sub>50</sub> in vivo. The results of this study shall contribute towards optimizing antitoxin production for BoNT/A1, which is essential for emergency preparedness and response.
Na, Domin,Ryu, Jaeil,Hong, Suk-Joo,Hong, Sun Hwa,Yoon, Min A,Ahn, Kyung-Sik,Kang, Chang Ho,Kim, Baek Hyun Korean Society of Magnetic Resonance in Medicine 2016 Investigative Magnetic Resonance Imaging Vol.20 No.2
Purpose: To analyze the feasibility of three-dimensional (3D) diffusion-weighted (DW) PSIF (reversed FISP [fast imaging with steady-state free precession]) sequence in order to evaluate peripheral nerves in the elbow. Materials and Methods: Ten normal, asymptomatic volunteers were enrolled (6 men, 4 women, mean age 27.9 years). The following sequences of magnetic resonance images (MRI) of the elbow were obtained using a 3.0-T machine: 3D DW PSIF, 3D T2 SPACE (sampling perfection with application optimized contrasts using different flip angle evolution) with SPAIR (spectral adiabatic inversion recovery) and 2D T2 TSE (turbo spin echo) with modified Dixon (m-Dixon) sequence. Two observers used a 5-point grading system to analyze the image quality of the ulnar, median, and radial nerves. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of each nerve were measured. We compared 3D DW PSIF images with other sequences using the Wilcoxon-signed rank test and Friedman test. Inter-observer agreement was measured using intraclass correlation coefficient (ICC) analysis. Results: The mean 5-point scores of radial, median, and ulnar nerves in 3D DW PSIF (3.9/4.2/4.5, respectively) were higher than those in 3D T2 SPACE SPAIR (1.9/2.8/2.8) and 2D T2 TSE m-Dixon (1.7/2.8/2.9) sequences (P < 0.05). The mean SNR in 3D DW PSIF was lower than 3D T2 SPACE SPAIR, but there was no difference between 3D DW PSIF and 2D T2 TSE m-Dixon in all of the three nerves. The mean CNR in 3D DW PSIF was lower than 3D T2 SPACE SPAIR and 2D T2 TSE m-Dixon in the median and ulnar nerves, but no difference among the three sequences in the radial nerve. Conclusion: The three-dimensional DW PSIF sequence may be feasible to evaluate the peripheral nerves around the elbow in MR imaging. However, further optimization of the image quality (SNR, CNR) is required.