Identification of an Immuno-dominant Small Antigen in Salmonella typhimurium

Jong Earn Yu,Young Hee Kim,Ho Young Kang 한국생명과학회 2005 한국생명과학회 심포지움 Vol.44 No.-

Contribution of antigenicity by Lpp, a Salmonella surface antigen in the immune response

Jong Earn Yu,Young Hee Kim,Seong Kug Eo,John Hwa Lee,Ho Young Kang 한국생명과학회 2005 한국생명과학회 심포지움 Vol.45 No.-

Roles of Glyceraldehyde-3-Phosphate Dehydrogenase in Edwardsiella tarda Pathogenesis

Jong Earn Yu(유종언),Young Eun Oh(오영은),Tae Ho Lee(이태호),Ho Young Kang(강호영) 한국생명과학회 2010 생명과학회지 Vol.20 No.12

Edwardsiella tarda는 그람 음성의 장내세균과의 주요 어병세균으로 어류에 edwardsiellosis를 유발하는 전신감염성 병원체이다. 최근 병원성 세균의 외막 단백질들은 세균성 감염에 있어서 숙주와 반응하여 면역반응을 유도하는 것으로 여겨져 연구가 되고 있다. 일본의 연구팀은 어류에서 에드워드병의 원인체인 E. tarda의 37 kDa 단백질이 넙치에서 높은 항원성을 제시하는 것을 보고하였다. 또한 그 연구자들은 37 kDa 단백질의 N-말단 아미노산 서열이 GAPDH와 대응하는 것을 밝혔다. 본 연구에서는 다른 세균에서 알려진 N-말단 서열을 기반으로 primer를 제작하여 이에 상응하는 E. tarda DNA를 증폭하고 클로닝하였다. 이 DNA단편의 염기서열은 예상한 바와 같이 세균의 GAPDH유전자인 gapA와 높은 상동성이 있고, E. tarda GAPDH (etGAPDH)의 아미노산 서열은 다른 장내세균의 GAPDH와 70% 이상의 상동성을 보이는 것을 확인하였다. E. tarda의 외막단백질에 특이적으로 반응하는 항체를 이용하여 E. tarda의 GAPDH가 외막에 존재한다는 것을 증명하였고, gapA의 염기서열을 바탕으로하여 재조합 GAPDH를 과발현 시켰다. 과발현된 재조합단백질 GAPDH는 GAPDH 특이적인 항체를 제조하는데 사용되었고, 또한 넙치에 면역시켜 단일 단백질 백신으로서의 활용도를 모색하였다. 비록 재조합 GAPDH가 면역된 넙치에서 GAPDH에 특이적인 항체가 증가하였음에도 불구하고, E. tarda로 공격실험을 하였을 때 면역된 넙치의 생존율이 12.5%로 측정되어 면역된 그룹과 면역되지 않은 그룹간에 큰 차이가 없는 것이 확인되었다. A research group demonstrated that the 37 kDA protein of Edwardsiella tarda, a causing causative agent of edwardsiellosis in fish, exhibited high antigenicity in Japanese flounder. The research group also showed that the N-terminus amino acid sequences of the 37 kDa protein were mapped to the N-terminus of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Using degenerated primer sets based on the known N-terminus sequence, the corresponding E. tarda DNA was amplified and cloned. The nucleotide sequences of the cloned gene revealed high homology with a bacterial gene for GAPDH, as we was expected. The amino acid sequence of E. tarda GAPDH (etGAPDH) revealed a <70% similarity with GAPDH proteins in other Enterobacteriaceae. With the application of artificial protein overexpression system in Escherichia coli, the recombinant etGAPDH (rGAPDH) was produced and purified. In this study, Using the purified rGAPDH, the etGAPDH specific polyclonal antibody has been was generated using the purified rGAPDHin this study. The immunoblotting analyses demonstrated that the location of the GAPDH protein is located with the association of is associated with the envelops of E. tarda. The rGAPDH was administrated into Japanese flounder via IP route for evaluation of the protective ability. Although the specific antibody titer against etGAPDH was increased about 3-fold after 4 weeks post-vaccination, the survival rates of vaccinated Japanese flounder and the control group with wild type E. tarda was were 12.5% and 0%, respectively. Our results indicated that rGAPDH is immunoreactive antigen but that it will not generate protective immunity in Japanese flounder.

Effects of Salt Concentration on Motility and Expression of Flagellin Genes in the Fish Pathogen Edwardsiella tarda

유종언(Jong Earn Yu),박준모(Junmo Park),강호영(Ho Young Kang) 한국생명과학회 2011 생명과학회지 Vol.21 No.10

염농도에 따른 E. tarda CK41의 운동성을 알아보기 위하여 1.0%와 3.5%의 염농도를 가지는 운동성 측정 배지에서 집락의 변화를 관찰한 결과, 3.5% 염농도 조건에서 운동성이 감소하는 것을 확인할 수 있었다. 1.0%과 3.5% 염농도 조건에서의 생육도를 측정해본 결과 각 염농도 조건에 따른 생균수의 차이는 매우 적은 것으로 보아, 높은 염농도에서의 운동성의 감소는 생육정체가 아닌 실질적인 운동성의 차이에 의함을 알 수 있었다. 이러한 염농도에 의한 운동성의 차이가 편모에 의한 것인지를 알아보기 위하여 투과 전자 현미경으로 형태학적 관찰을 해본 결과, 3.5% 염농도에서는 편모의 형성이 되지 않음을 확인하였다. E. tarda는 PFAD와 FDP 두개의 편모 유전자를 가지며 이들간의 아미노산 상동률은 93%로 높은 편이다. 편모의 발현양의 확인을 위하여 PFAD 특이적인 다클론성 항체를 제작하기 위하여, PFAD를 과발현시키는 재조합 플라스미드 pBP793을 구축하여 대장균 발현시스템으로 발현시켜 정제한 후, 토끼에서 면역반응을 유도하여 특이항체를 제작하였다. PFAD 특이적인 다클론성항체를 이용한 immunoblot assay 결과, 3.5% 염농도 조건에서 배양한 E. tarda CK41의 경우 1.0% 염농도에서 보다 반응하는 면역 활성 단백질 밴드가 낮은 것으로 측정되었다. 이러한 결과를 종합하여 볼 때, 염농도가 높은 해수환경에서의 운동성의 감소는 E. tarda CK41의 편모 단백질이 제대로 발현되지 않아 기능적인 편모의 형성이 이루어지지 않는다는 것을 예증하고 있다. 향후 연구에서 어떠한 메카니즘에 의해 염농도가 flagellin의 발현을 조절하는지를 밝힐 필요가 있다. E. tarda, a fish pathogen, can survive in seawater under relatively high salt conditions as well as in fish under physiological salt conditions. Bacterial growth under different salt concentrations may influence the expression of genes involved in bacterial structure and physiology. The growth rate of E. tarda culture in high salt (3.5% NaCl) was similar to that in low salt (1.0% NaCl, physiological salt concentration). Interestingly, the strain moved much faster in low salt conditions than in high salt conditions. Electron microscopic observation demonstrated that the bacterial cells grown in high salt had less or no flagellation. Obvious flagellation was observed in the parental strain E. tarda CK41 grown in low-salt condition. Two putative genes coding flagellin were identified in the E. tarda genome sequences. The amino acid sequence comparison of each gene revealed 93% identities. A flagellin gene was PCR amplified and cloned into a cloning vector. Using an E. coli protein expression system, a part of flagellin protein was overexpressed. Using the purified protein, an anti-flagellin antibody was raised in the rabbit. Immunoblot analyses with flagellin specific antibody demonstrated that E. tarda CK41 expressed falgellin in low salt conditions, which is consistent with the results seen in motility assay and microscopic observation. This is the first report of salt regulated flagella expression in E. tarda.

A Good Outcome for a Case of Chronic Pneumonitis of Infancy

Kim, Jong Min,Kwon, Seung Yeon,Kim, Eun Soo,Sohn, Myung Hyun,Kim, Kyu Earn,Kim, Myung Joon,Cho, Sang-Ho Yonsei University, College of Medicine 2007 Yonsei medical journal Vol.48 No.5

<P>Chronic pneumonitis of infancy (CPI) is a very rare interstitial lung disease. Its pathological features differ from other types of interstitial pneumonia that occur in adults and children. The mortality rate of CPI is high, even with treatment. We report a case of a 3 month old girl diagnosed with CPI after an open lung biopsy who improved after proper treatment.</P>

고해상도 흉부 단층 촬영 결과를 통한 감염 후 폐색성 세기관지염의 임상적 접근

김종덕 ( Jong Deok Kim ),한윤기 ( Yoon Ki Han ),김경원 ( Kyung Won Kim ),이미정 ( Mi Jung Lee ),손명현 ( Myung Hyun Sohn ),김규언 ( Kyu Earn Kim ) 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 2012 소아알레르기 및 호흡기학회지 Vol.22 No.4

목 적 : 폐색성 세기관지염은 만성 폐쇄성 폐 질환의 한 형태로 소아에서는 여러 원인에 의한 폐 감염 후 발생하는 경우가 가장 많은 것으로 보고되고 있다. 본 연구는 폐색성세기관지염 진단에 유용하게 사용되는 고해상도 흉부 단층 촬영 결과를 토대로 임상 양상을 조사하여 감염 후 폐색성세기관지염을 진단하고 이들의 임상적 특징과 원인, 방사선 소견 등을 조사하고 이를 대조군과 비교하여 향후 진단과 치료적 접근에 도움을 주고자 시행되었다. 방 법: 2007년 1월부터 2011년 3월까지 연세대학교 세브란스병원 소아청소년과에서 시행한 모든 고해상도 흉부 단층 촬영의 결과들을 확인하여 폐색성 세기관지염에 특징적인 소견을 동반한 결과를 보인 환아들의 영상 소견 및 임상 증상을 재확인하였고 이를 통해 감염 후 폐색성 세기관지염으로 진단된 52명을 대상으로 후향적 조사를 시행하였다. 또한 급성 호흡기 증상을 주소로 내원하여 고해상도 흉부 단층 촬영을 시행한 환아에서 단순 하기도 감염으로 진단된 65명을 대조군으로 설정하여 내원 당시의 임상적 특징들을 감염 후 폐색성 세기관지염군과 비교하였다. 결 과 : 감염 후 폐색성 세기관지염 진단 당시의 평균 연령은 41.3±5.9개월이었고, 초기 감염 시 평균 연령은 14.6±2.8개월이었다. 이에 비해 대조군의 진단 당시의 평균 연령은 57.1±5.1개월이었고, 초기 감염 시 평균 연령은 50.9±5.0개월로 통계적으로 유의한 차이가 있음을 확인하였다. 또한 초감염 후 진단 당시까지 급성 하기도 감염으로 입원 치료 받은 빈도수에서 폐색성 세기관지군이 대조군에 비해 상대적으로 빈번하게 노출되었음을 확인하였다. 결 론 : 본 연구에서는 지속되는 호흡기 증상을 주소로 내원한 환아에서 반복적인 호흡기 감염의 입원력이 있고, 2세미만의 어린 연령에서 하기도 감염의 과거력이 있는 경우, 고해상도 흉부 단층 촬영을 통한 폐색성 세기관지염의 진단적 접근을 시행할 필요가 있으며 진단 후 지속적인 차후 관리가 이루어져야 한다고 생각한다. Purpose : Bronchiolitis obliterans (BO) is a type of chronic obstructive lung disease. In children, the most common presentation is a post infectious origin. The aim of this study was to determine the clinical features, etiology, and radiologic findings of post infectious BO, which was diagnosed by high resolution computed tomography (HRCT). The results were then compared to those of the control group. Methods : We diagnosed 52 children with BO by a review of all performed HRCT findings at Severance Children`s Hospital between January 2007 and March 2011. A retrospective study was conducted to define the clinical features, etiology, symptoms, and radiologic findings of post infectious BO. Sixty-five children who were diagnosed with lower respiratory infection confirmed by HRCT were defined as the control group for comparison with post infectious BO patients. Results : The mean age of the patients at the time of diagnosis with BO was 41.3±5.9 months, and that of the control group was 57.1±5.1 months. The initial insult age of BO patients was 14.6±2.8 months, and that of the control group was 50.9±5.0 months. The frequency of previous admissions due to lower respiratory infection was relatively higher in BO patients than the control group. Conclusion : The results suggest that the development of post infectious BO should be early suspected and HRCT should be performed in children with a recurrent admission history of lower respiratory infection, especially in those younger than 2 years of age. Furthermore, continued management of prevent additional infection should be conducted. [Pediatr Allergy Respir Dis(Korea) 2012;22:397-403]

한국 소아에서 폐색성 세기관지염에 관한 다기관 연구

홍수종(Soo Jong Hong),김봉성(Bong Seong Kim),안강모(Kang Mo Ahn),이상일(Sang Il Lee),김규언(Kyu Earn Kim),이기영(Ki Young Lee),임대현(Dae Hyun Lim),손병관(Byong Kwan Son) 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 2002 소아알레르기 및 호흡기학회지 Vol.12 No.2

N/A Background: Bronchiolitis and bronchiolitis obliterans(BO) are an inflammatory lung diseases that primarily affects the small conducting airways. There are few data regarding the epidemiology, pathophysiology, long term sequelae, and the therapy of bronchiolitis obliterans. The aim of the study is to determine the clinical profile, etiology, high resolution computed tomography(HRCT) findings and response to treatment in Korean children diagnosed with BO. Methods: We performed a retrospective study to define the clinical course, HRCT findinding, etiologic agents, initial and follow-up symptom score(Denver) , and response to intravenous corticosteroid of 34 children with BO in four university hospitals between February 1995 and February 2000. Results: Median age of diagnosis of BO was 17.5 months(3 to 79 months) , that of initial infectious insult was 11 months(3 to 79 months), and median duration of BO diagnosed after acute illness was 1 month(1 to 32 months). Clinical manifestations are prolonged/recurrent cough(97%), sputum(85%), respiratory difficulty(65%), wheeze(59%), fever(44%), and exertional dyspnea(18%). HRCT findings included mosaic perfusion(65%), bronchial wall thickening(62%), atelectasis(47%), hyperinflation(44%), varying degrees of bronchiectasis(35%), and ground glass opacity(25%). Thirty two cases were associated with infections and most common infectious agents was adenovirus(n=15, 44% ). Other infections were influenza virus (n=3, 9%), mycoplasma(n=2, 6%) , respiratory syncytial virus(n=1, 3%), and parainfluenza virus(n=1, 3%). Initial symptom scores were higher in intravenous corticosteroid treated group than untreated group, and follow-up scores were decreased in both groups. Conclusion: Most cases associated with infection and Adenovirus was the most common cause of BO. Corticosteroid treatment during the acute and chronic phases may improve the functional status of BO patients.

한국 소아알레르기 질환의 유병률

홍수종 ( Soo Jong Hong ),안강모 ( Kang Mo Ahn ),이수영 ( Soo Young Lee ),김규언 ( Kyu Earn Kim ) 대한소아알레르기 및 호흡기학회 2008 소아알레르기 및 호흡기학회지 Vol.18 No.1

Attitudes and preferences of consumers toward food allergy labeling practices by diagnosis of food allergies

Se-young Ju,Jong-Hwan Park,Tong-Kyoung Kwak,Kyu-earn Kim 한국영양학회 2015 Nutrition Research and Practice Vol.9 No.5

BACKGROUND/OBJECTIVES: The objective of this study was to investigate food allergens and prevalence rates of food allergies, followed by comparison of consumer attitudes and preferences regarding food allergy labeling by diagnosis of food allergies. SUBJECTS/METHODS: A total of 543 individuals living in Seoul and Gyeonggi area participated in the survey from October 15 to 22 in 2013. RESULTS: The results show that the prevalence of doctor-diagnosed food allergies was 17.5%, whereas 6.4% of respondents self-reported food allergies. The most common allergens of doctor-diagnosed and self-reported food allergy respondents were peaches (30.3%) and eggs (33.3%), respectively, followed by peanuts, cow’s milk, and crab. Regarding consumer attitudes toward food labeling, checking food allergens as an item was only significantly different between allergic and non-allergic respondents among all five items (P < 0.001). All respondents reported that all six items (bold font, font color, box frame, warning statement, front label, and addition of potential allergens) were necessary for an improved food allergen labeling system. PLSR analysis determined that the doctor-diagnosed group and checking of food allergens were positively correlated, whereas the non-allergy group was more concerned with checking product brands. CONCLUSIONS: An effective food labeling system is very important for health protection of allergic consumers. Additionally, government agencies must develop policies regarding prevalence of food allergies in Korea. Based on this information, the food industry and government agencies should provide clear and accurate food labeling practices for consumers.

Development of Biomarkers against Low Pathogenic Avian Influenza Virus (H9N2/MS96) Utilizing Variable Lymphocyte Receptors (VLRs) of Inshore Hagfish (Eptatretus burgeri)

Se Pyeong Im,Jong Earn Yu,Jung Seok Lee,Ho Bin Jang,Seong Bin Park,Si Won Kim,Tae Sung Jung 대한수의학회 2014 대한수의학회 학술대회발표집 Vol.2014 No.-