간흡충 성충의 조직항원 부위와 충체 추출물의 항원성

이옥란,이민구 순천향의학연구소 1997 Journal of Soonchunhyang Medical Science Vol.3 No.2

간흡충(Clonorchis sinensis)의 수용성추출물의 항원성과 충체조직의 부위별 조직항원의 황원성을 비교함으로써 수용성항원의 항원성을 주도하는 체조직항원부위를 알아보고자 이 실험을 시도하였다. 간흡충의 수용성항원은 충체 추출물(조항원)을 DEAE-anion exchange chromatography로 부분 정제한 분리하원(D1, D2, D3, D4 및 D5)을 사용하였고, 이들 수용성항원은 SDS-PAGE로 각 항원의 분자량별 단백분획상과 EITB로 단백분획의 항원성을 비교하였다. 조직항원의 항원성은 면역황금 표지법으로 부위별 조직항원을 검사하였다. 미세 조직항원의 면역반응 관찰에는 간흡충 감염 토끼 혈청, D3면역 토끼혈청 및 D5면역 토끼혈청을 사용하였다. 간흡충의 수용성항원과 분리항원들은 공통분획들이 관찰되었고 일부 분획들은 염색상 농도 차이를 보였다. 이들 수용성항원의 단백분획들 대부분이 항체와 결합한 것으로 보아 충체 단백물질 대부분이 항원성이 있음을 알 수 있었다. 모든 항원에서 항체와 강하게 결합된 공통 분획은 27 kDa, 16 kDa및 14 kDa분획이었다. 수용성항원과 감염 흰주혈청을 사용한 ELISA에서 D5항원은 감염초기(감염2주)를 제외한 전 감염기간 중 강력한 반응을 보였고, D1항원은 감염초기(2주)에 강한 항원성을 보였다. 면역황금표지법 상 간흡충의 표피, 장상피, 배설낭 및 난황선과 항혈청들(감염혈청, D3면역혈청및 D5면역혈청)의 반응은 강도의 차이는 있으나 항원이 있는 조직들이었다. 전 조직은 D5면역혈청반응에서 강한 항원성을 보였고 D2및 D3면역혈청에서 약한 반응을 보였다. D5면역혈청을 반응시켰을 때 난황선은 많은 황금입자가 표지되어 항원단백질이 풍부한 조직부위로 보이나 감염혈청과 조직의 면역반응에서는 난황선의 항원성은 강하지 않았다. 면역황금표지법 상 간흡충의 표피, 장상피, 배설낭 및 난황선과 항혈청들(감염혈청, D3면역혈청및 D5면역혈청)의 반응은 강도의 차이는 있으나 항원이 있는 조직들이었다. 전 조직은 D5면역혈청반응에서 강한 항원성을 보였고 D2및 D3면역혈청에서 약한 반응을 보였다. D5면역혈청을 반응시켰을 때 난황선은 많은 황금입자가 표지되어 하원단백질이 풍부한 조직부위로 보이나 감염혈청과 조직의 면역반응에서는 난황선의 항원성은 강하지 않았다. 결론적으로 간흡충의 충체추출 수용성항원은 간흡충충체 항원단백질의 혼합체로 구성되었으나 감염혈청과 면역혈청에서 강한 항체결합을 한 장상피 및 배설낭이 주 항원성 단백질함유조직으로 보이며, 이 중 가장 강한 항원성을 보인 장조직은 수용성항원의 항원성을 주도하는 조직항원부위로 생각한다. The major antigenic site in the tissue of Clonorchis sinensis were observed and compared with antigenicity of the worm extracted soluble antigens. The worm extracted crude antigen(CSWA) was separated into five protein fractions(seperate antigens: D1, D2, D3, D4 and D5) by DEAE anion exchange chromatography. These antigens were analized with SDS-PAGE(SDS-polyacrylamide gel electrophoresis) and the EITB(enzyme linked immunotransfer blot). Antigenicity of the crude and separated five antigens were evaluated by ELISA(enzyme-linked immunosorbent assay) reacted with sera from rat infected with Clonorchis sinensis. The antigenicity in tissue sites of Clonorchis sinensis was observed with electronmicroscopy by immunogold labeling method using three anti rabbit sera (the infected serum with Clonorchis sinensis, the D3 and the D5 immune rabbit sera). The results were as follows: By EITB, the most protein fractions on SDS-PAGE were binded to infected rat sera, and the strongly binding fractions against in all antigens were molecular weight of 27 kDa, 16 kDa and 14 kDa. In antigenicity estimated by the OD value of ELISA, the D5 was revealed stronger than those of the others. Statistically, the antigenicity of the D5 was stronger(p<0.001) than that of other separated antigens(p<0.05 ~ p<0.001) from 4 weeks to 20 weeks after infection. In the ultrastructural tissues, the tegumental syncytium, the intestinal epithelium, the excretory bladder and the vitellaria of Clonorchis sinensis were labeled with different density of the immunogold particles against all three antisera. The immunogold particles of a large numbers were labeled on the intestinal epithelium and the excretory bladder, but in the tegument and the vitellaria were labeled scarecely. Much more numbers of immunogold particles were labeled in D5 immune serum than other antisera, and especially, a large number of immunogold particles were labeled in intestinal epithelium. Summarizing the above results, although the antigenicity of Clonorchis sinensis was shown in all the tissues(tegument, intestine, excretory bladder and vitellaria), the major antigenic site was to be the intestinal region.

Epidemiological studies of Clonorchis sinensis in the coastal areas of Kyungpook, Korea

Ahn,Sung Hoon,Lee,Soo Hyeong,Joo,Chong Yoon 啓明大學校 醫科大學 1996 계명의대학술지 Vol.15 No.4

In order to determine the epidemiological patterns of the liver fluke, the infection rates for Clonorchis larvae in the snails and fish hosts, and the prevalence of C. sinensis among the residents in the coastal areas of Kyungpook Province were studied from April, 1992 to September, 1993. Four snail habitats were found in the river Hyungsan and come streams. One area had snails infected with Clonorchis cercaria, but the proportion of infected snails was very low, the average rates varying from zero to 0.081 percent. Of the 19 species of fish, 3 species were infected with the encysted larvae of C. sinensis. Infection rates varied greatly by the fish species, and ranged from 6.7 percent in P. herzi to 88.9 percent in G. artomaculatus. In the intensity of infection with C. sinensis, G. atromaculatus was the most heavily infected species, the average number of cysts per gram of the fish flesh being 72.2 followed by P. herzi with 53.0 cysts. The infection rate for C. sinensis among the residents was relatively high, being found to be 5.9 percent with a prevalence of 7.0 percent in males and 5.1percent in females. the difference in the rate of infection between males and females is found to be statistically significant(t>2). For the quantitative analysis of epidemiology of C. sinensis, a special type of simple catalytic model was applied to the sex and age infection rates revealed by stool examination for Clonorchis eggs. The simple catalytic curve for males was y=0.21(1-e-0.021t), and females was y=0.19(1-e-0.013t). From these equations the force of infection of 2.1 and 1.3 effective infections giving positive stool examination per 1,000 residents annually was suggested. And the asymptote is at about 21.0 percent of males and 19.0 percent of females, the estimated values of the limit of possible positive cases in the residents. For the analysis of the age prevalence patterns of Clonorchis positives applied the two-stage catalytic model, and the equation is y=0.511(e-0.0093t-e-0.0275t). for the analysis of the intensity of infection, the frequency distribution by the Clonorchis egg counts was calculated as well as the cumulative percentage to the total number of positive cases, and the regression equation is y=1.07+1.50 log X. This study indicate that endemic foci of C. sinensis exist in the coastal areas of Kyungpook Province and the prevalence of the liver fluke among the residents is still high, and that the infection rates with Clonorchis larvae in the fresh-water fish and snails varied greatly by the species of fish and its habitats.

간흡충 감염가토의 담관에 대한 병리조직학적 변화 및 Lectin 반응에 관한 연구

양웅석(Ung Suk Yang),조몽(Mong Cho),백태현(Tae Hyun Paik),김상현(Sang Hyun Kim),주형준(Hyung Jun Joo),이동필(Dong Phil Lee),송근암(Geun Ahm Song),문한규(Han Gyu Moon) 대한소화기학회 1993 대한소화기학회지 Vol.25 No.6

N/A Histopahological and histochemical studies were done in the bile ducts of clonorchis sinensis (CS) infested rabbits. The duration of clonorchis sinensis infestation in the rabbits was between 12 and 17 months and the number of clonorchis sinensis metacercariae which were infested orally was 50-150 per rabbit. Hematoxylin-eosin and avidin-biotin peroxidase complex lectin staining methods were used for this study. Pathological changes of the bile ducts of CS-infested rabbits were papillarv and adenomatous hyperplasia, goblet cells and mucus cells rnetaplasia, ductal wall fibrosis and infitra tion of eosinophils in the ductal wall. These pathological changes were noted both in peripheral and central protion of bile duct. The pathological changes of hyperplasia and metaplasia of ductal epithelium were more marked in the deep gland than surface one, and dysplasia of the epithelium was more marked in the suface gland than deep one. In clonorchis sinensis infested rabbits, the lectins of Soybean agglutinin (SBA), Dolichos biflours agglutinin (DBA) and Ulex europaeus agglutinin (UEA I) were labelled predominant]y in the luminal suface, supranuclear protion of epithelium, goblet cells, hyperplastic cells and metaplastic cells of deep gland . SBA and DBA were labelled weakly to intermediate and UEA I was labelled weakl y to strong, but Peanut agglutnin (PNA) was ]abelled very weakly to those sites in CS-infested rabbits. In non CS infested rabbits, lectins of SBA, DBA, and UEA I were only weakly labelled in the luminal surface of epithelium. In this study, IJEA i which binds specifically glycoprotein of fucose was more pronounced in rabbits infested with CS and so it is postulated that increased activity of fucosyl transferase should be associated with CS infestation.

간흡충에 감염된 실험쥐 담관 섬유모세포의 미세구조적 변화

김수진,민병훈,Kim, Soo-Jin,Min, Byoung-Hoon 한국현미경학회 2004 Applied microscopy Vol.34 No.2

동물의 결합조직에 분포하고 있는 섬유모세포 (fibroblast)는 결합조직을 구성하는 세포의 한 종류로서 세포질 돌기들이 잘 발달된 형태적 특징이 있는 것으로 실험쥐 담관의 경우 간흡충 등의 기생충에 의하여 물리, 화학적 상해를 받았을 때 세포변이가 유발될 뿐만 아니라, 담관 암세포로 전이되기도 하는 것으로 알려져 있다. 따라서 저자 등은 실험쥐의 담관이 기생충에 의한 상해를 받았을 때 섬유모세포의 세포 표면과 세포질의 변화를 알아보고자 실험쥐 담관에서 섬유모세포를 분리하여 전자현미경으로 확인하고 다음과 같은 결과를 얻었다. 대조군 실험쥐 담관의 섬유모세포들은 일반적인 형태로 세포돌기, 세포표면 및 세포질을 구성하고 있었으나 간흡충 감염군 실험쥐 담관의 섬유모세포는 미세소관에 의한 세포질 돌기들이 다수 발달하고 다양한 종류의 포낭형 조면소포체 그리고 세포질에 전자밀도가 높은 다양한 액포, 높은 밀도의 리보좀을 포함하는 조면소포체, 다양한 형태의 과립 및 많은 수의 미세섬유가 관찰되는 형태적 변화가 관찰되었다. 간흡충에 감염된 담관의 섬유모세포는 간흡충에 의하여 상해 받은 세포가 물리화학적 자극에 의한 적응으로 단백질 합성이 증가하며 multi-vesicular 형태의 Golgi복합체가 생성되고, 세포질돌기 형성하는 것으로 확인되었다. 세포질에 광범위하게 분포하는 multi-vesicle은 당말단인 sialic acid를 포함하고 세포내에서 세포표면의 미세융모에 이르기까지 이동하는 것으로 확인되었다. 이상의 결과로 간흡충 감염 실험쥐로부터 분리된 섬유모세포는 actin단백으로 구성된 세포돌기가 잘 발달하고, 세포내 조면소포체에서 형성된 단백질이 Golgi복합체에서 당말단인 sialic acid로 전환되어 세포표면에 분포하게 된다. 이는 간흡충 감염으로 물리 화학적 자극 자극받은 섬유세포가 미세구조적 변화를 유발하는 것으로 확인되었다. In this study, ultrastructural change of the bile duct fibroblast at infected rat with Clonorchis sinensis, and the distribution of lectin receptors and actin protein in cultured bile duct infected with Clonorchis sinensis. It explored using colloidal gold label complex with lectin WGA purified from wheat germ (Triticum vulgaris) and anti actin antibody purified actin (43 kDa) isolated from chicken back muscle. The lectin WGA with protein A gold complex labeled sections of the cultured fibroblast revealed gold particles specifically distributed on the multi vesicular form Golgi complex and cell surface of the fibroblast. The actin antibody with protein A gold complex labeled sections of the cultured fibroblast revealed gold particles specifically distributed on the cytoplasm of the fibroblast. Labeling of cultured fibroblast in rat bile duct infected with Clonorchis sinensis was then quantified and compared to that of cultured Fibroblast in Rat Bile duct. These results indicate that lectin WGA receptors are located in the multi vesicular form Golgi complex in the cytoplasm to the cytoplasmic process of the Rat bile duct fibroblast infected with Clonorchis sinensis. Therefore, the GlcNAc and NeuNac regions on the cell surface and cytoplasmic process appear to be functionally associated with cell-recognition and protection from other cell of the tissue, and linked with secretion and exocytosis of the fibroblst cytoplasm. GlcNAc and NeuNAc product in the multi vesicular form Golgi complex then it is transported to cell surface. Actin protein is many appears that infected fibroblast rather than normal fibroblast. The fibroblast of infected with Clonorchis sinensis are against of the physical and chemical stimulation. Then development of cytoplasmic process is relative some stimulation.

Epidemiological Studies of Clonorchis sinensis in the Vicinity of Cheongdo River, Kyungpook, Korea

Shin, Dong Hak,Kwak, Ki Woo,Joo, Chong Yoon 啓明大學校 醫科大學 1999 계명의대학술지 Vol.18 No.2

In order to determine the epidemiological pattern of the liver fluke, the infection rates and intensity for the metacercarial larvae of C. sinensis in the fish hosts, and the prevalence of the liver fluke among the residents in the vicinity of the Cheongdo River, Kyungpook Province Korea were studied from June, 1993 to September, 1994. Fourteen species of fresh-water fish caught in the river were examined for the encysted larvae of C. sinensis. Of these, 6 species were infected with the metacercariae of C. sinensis. Infection rates varied greatly by fish species and ranged from 16.7 per cent in P. herzi to 80.0 per cent in P. parva. As for the intensity of Clonorchis infection, P. parva was the most heavily infected species, with an average number varying from 8.8 to 11.0 cysts. The prevalence of C. sinensis among the residents in the vicinity of the Cheongdo River was relatively high, being found to be 21.0 per cent with an infection rate of 23.47 per cent in males and 17.86 per cent in females. The difference of infection rate between males and females is found to be statistically significant (t>2). For the analysis of age pre valence patterns of Clonorchis positives applied the two-stage catalytic model, and the equation was y=1.2(e(-0.015t)-e(-0.022t)) From this equation the force of infection was annually 15 per 1,000 susceptibles and the disappearance rate from positive to negative cases was annually 22 per 1,000 Clonorchis positive in this population. For the analysis of intensity of infection, the frequency distribution by the Clonorchis egg counts was calculated as well as the cumulative percentage to the total number of positive cases, and the regression equation was y=1.51+1.30 logX. These figures indicated that most cases of the residents with clonorchiasis were found to be lightly infected. This study indicate that endemic foci of C. sinensis exist in the vicinity of the Cheongdo River Cheongdo and the prevalence of the liver fluke among the residents is still high.

간흡충: 충체 및 대사성 항원의 특성분석 (1)항원투여 마우스 비장조직에 대한 면역조직화학적 연구

송강원,주난영,류장근,양용석 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.2

저자들은 마우스를 실험모델로 하여 간흡충의 항원을 투여 했을 때 비장조직에 대한 CD3, CD4 및 CD8 모노클로날 항체의 반응 여부를 알아보고자 하였다. 즉, 간흡충에 대한 세포면역학적인 특성을 규명고자 하였으며 특히 비장 조직에 대한 phenotype을 관찰한 결과 다음과 같은 결과를 얻었다. 간흡충의 조항원을 면역증강제와 함께 복강 투여한 다음 일정 기간 후에 비장조직을 Avidin-biotin complex 면역조직염색을 실시한 결과 CD3에서 강한 양성 반응을 나타냈고 CD4와 CD8에서는 약한 반응을 나타냈다. 조직부위를 보면 피막, 혈관, 임파관, 백수부위와 림프구 및 대식세포의 세포막에서 양성반응을 보였다. The authors inquired into what reactions comprise the response of mice(as a model) CD3, CD4 and CD8 monoclonal antibodies in spleen tissue when injected intraperitoneally by antigens of Clonorchis sinensis. The author is objective was focused on investigating the property of cellular immunity for liver fluke. In particular, the results of having examined the phenotype of the tissue of spleen were revealed as follows: a certain length of time after having been intraperitoneally injected with antigens of Clonorchis sinensis and Freund's adjuvant, the tissue of spleen was embedded and immunohistochemically stained by the avidin-biotin complex method. A strong reaction in response to CD3, while a feeble reaction resulted from CD4 and CD8. The tissue region showed a positive reaction to all antibodies, especially from capsules, vascular areas, white pulps and membrane of blood cells.

평택천 주변의 담수어에서 간흡충 피낭유충의 검출

전계식 한국환경생물학회 1998 환경생물 : 환경생물학회지 Vol.16 No.4

1998년 6월 하순 평택천 주변인 경기도 평택군 오성면 당거리와 길음리 및 현덕면 신대리에서 포획한 담수어를 실험실에 운반하여 인공소화액(pepsin-HCI)으로 37℃ 배양기에서 1시간 동안 소화시킨 다음 입체현미경으로 피낭유충을 검경하였다. 조사한 담수어 7종 74마리 가운데 간흡충 피낭유충은 4종 26마리에서 총 128개체가 검출되었다. 간흡충의 피낭유충은 참붕어 Pseudorasbora parva는 11마리 중 9마리(81.8%)에서 감염 어체당 평균 6.9개체가 검출되었으며 몰개 Gnathopogon coreanus는 15마리 중 6마리(40.0%)에서 평균 5.5개체, 처리 Cultriculus eigenmani는 24마리 중 10마리(41.7%)에서 평균 3.2개체 그리고 붕어 Carassius carassius는 3마리 중 1마리(33.3%)에서 평균 0.3개체가 검출되었고 빙어 Hypomesus olidus, 밀어 Rhinogobius brunneus 및 미꾸리 Misgurnus anguilli-caudatus에서는 검출되지 않았다. 이상의 조사 결과로부터 평택호 주변 지역의 담수어 중에서 간흡충 피낭유충이 감염되어 있었고 참붕어는 다수 감염되어 있음이 확인되었다. In order to survey the infection status of Clonorchis sinensis metacercariae in freshwater fishes from Phungtack stream located in Kyunggin Province, a total of 74 fishes of 7 species were collected by a fish net on the late of June, 1998. They were brought to the laboratory and examined under the stereomicroscope after artificial digestion with pepsin-HCI solution in a 37℃ incubator for one hour. Clonorchis sinensis metacercariae was found from the total of 26 individuals of 4 species of freshwater fishes. The most frequently infected fishes were Pseudorasbora parva(81.8%), followed by Cultriculus eigenmani(41.7%), Gnathopogon coreanus(40.0%) and Carassius carassius(33.3%), respectively. The average number of metacercariae in P. parva, G. coreanus, C. eigenmani and C. carassius was 6.9, 5.5, 3.2, 0.3 respectively. From the results obtained in the survey, it was confirmed that freshwater fishes from Phungtack stream were infected with metacercariae of Clonorchis sinensis.

Transfer of Immunity by Peritoneal Cells, Spleen Cells and Sera in Golden Hamsters infected with Clonorchis sinensis

Kim, Jun Taek,Kong, Hyun Hee,Moon, Chu Hwan,Choi, Dong Wik 慶北大學校 醫科大學 1994 慶北醫大誌 Vol.35 No.2

肝吸蟲 成蟲의 代謝産物과 蟲體構成物의 footpad내 注射로 感作한 golden 햄스터의 腹腔細胞와 血淸 또는 脾臟細胞와 血淸을 同系 recipient 햄스터의 腹腔 내에 注入하였을 때 肝吸蟲 成蟲에 대한 免疫이 移入되는지 그 與否를 對照群 햄스터의 EpG, worm burden, 脾臟 當 plaque 形成細胞數를 基準으로 하여 對照群에서의 그 成績과 比較하였다. Challenge 感染 後, EpG는 對照群에서 感作群에 비하여 일찍, 많이 나타났으나, 肝吸蟲 成蟲의 回收率에서는 對照群과 感作群 사이에 有意的인 差異를 認定할 수 없었다. 脾臟에서 Plaque 形成細胞는 對照群 햄스터에서 검출되지 않았으나, 感作群에서는 相當數 檢出되었다. OBJECTIVE, MATERIALS AND METHODS : In order to review the consistency of passive transfer of the immunity against Clonorchis sinensis in golden hamsters, the donor hamsters were sensitized with two injections of the admixture of the metabolic products and/or the somatic constituents of C. sinensis and Freund's incomplete adjuvant into the footpads at 2 weeks' interval. The peritoneal exudate cells and sera and/or the spleen cells and sera were injected intraperitoneally to the recipient hamsters. Seven days after primary sensitization, the recipient hamsters were challenged with 50 metacercariae of C. sinensis. RESULTS : The results obtained were compared with those reported by Choi and Park(1987) and Choi et al. (1990), which showed significant reductions in the worm burdens. It was found that the eggs from control hamsters appeared earlier than those from the recipient groups, but there was no significant difference in the eggs per gram of feces and the worm burden of C. sinensis between the recipient and control groups. CONCLUSIONS : It is likely that these results show in part the variability of adoptive transfer of immunity against C. sinensis in golden hamsters.

경상남도 일부 농촌지역의 간흡충 감염에 대한 역학적 연구

주영희,오진경,공현주,손운목,김윤규,김정일,정갑열,신해림,Ju, Young-Hee,Oh, Jin-Kyoung,Kong, Hyun-Joo,Sohn, Woon-Mok,Kim, Yoon-Gyu,Kim, Jung-Il,Jung, Kap-Yeol,Shin, Hai-Rim 대한예방의학회 2005 Journal of Preventive Medicine and Public Health Vol.38 No.4

Objectives : To investigate the prevalence of Clonorchis Sinensis infestation and to determine the associated risk factors among a population in a part of the rural area of Gyeongsangnam-do, Korea, which is an area known to have a high mortality from liver cancer and a high infection of C. sinensis. Methods : The study populat ion consisted of those people who have lived in rural areas and who were over 40 years old. This study was performed in 5 areas during the period from 1999 to 200 3. Informed consents were obtained from the 2,381 study participants, and these people were interviewed about their life style habits with using the structured questionnaire that was administered by trained staff members. The subjects underwent blood sampling and their stool specimens were examined by using the Formalin-ether sedimentation technique. Results : Among the study subjects (N=2,381), the positivity for C. sinensis in the stool was 34.4% (95% CI=36.3-42.5); it was 39.4% (95% CI=36.3-42.5) in the males and 30.9% (95% CI=28.5-33.3) in the females . The positivity for C. sinensis was associated with current alcohol drinking (odds ratio=1.8, 95% CI: 1.5-2.1) and raw fish consumption (odds ratio 1.2, 95% CI: 0.9-1.6). Conclusion : The present study confirmed the high prevalence of C. sinensis in the study subjects. It is necessary to establish consistent medical management and education programs for the treatment and prevention of C. sinensis infestation in the rural inhabitants of Korea.

Clonorchis sinensis: Analysis Characterization of Somatic and Metabolic Antigen (II) Profile of the Worm, Excretory-secretory and Billis Antigen in C. Sinensis Infected Rabbit

Lee,Ji-Sook,Ryang,Yong-Suk,Cho,Yoon-Kyung THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1997 Journal of biomedical laboratory sciences Vol.3 No.2

가토에게 간흡충을 간염시키고 3개월 후 거살한 다음 간흡충의 충체, 간흡충의 분비배설액 그리고 간토의 담즙을 조항원으로 제조한 후 이에 대한 항원단백질의 구성물질을 분석하였다. 그리고 cysteine계 면역억제제인 E-64와 serine계 면역억제제인 PMSF를 첨가하였을 때 단백질 구성물질의 발현 양상을 관찰하였다. 실험적으로 가토에서 얻은 간흡충 성충의 조항원은 200-9 kDa의 범위에서 26개의 분획을 관찰하였으며, 간흡충 성충 조항원에 cysteine계 단백질분해효소 억제제인 E-64를 첨거하였을 때 잘 보존하여 200-9 kDa의 범위에서 29개의 분획이 관찰되었다. 그리고 간흡충을 감염시킨 가토의 담즙 조항원은 200-10 kDa의 범위에서 19개의 분획이 관찰되었으며, serine계 단백질분해효소인 PMSF를 첨거하였을 때 200-9 kDa의 범위에서 22개의 분획이 관찰되었으나, cysteine계 단백질분해효소인 E-64에서도 비슷한 양상을 보였다. 대조군인 건강 가토의 담즙 조항원은 200-10kDa의 범위에서 22개의 분획이 관찰되었으며, serine계 단백질분해효소 억제제인 PMAF를 첨가하였을 때 잘 보존하여 200-12kDa의 범위에서 23개의 분획이 관찰되었다. 앞으로 각 항원에 대한 면역학적 특징 및 단세포군 항체에 대한 구체적인 규명이 계속되어야 겠다. The authors characterized the proteins of the crude antigen obtained from Clonorchis sinensis worm and excretory-secretory and billis from rabbits, experimentally infected for 3 months. Protein composition was observed after adding a cysteine proteinase inhibitor E-64 and a serine proteinase inhibitor PMSF, respectively. SDS-PAGE of the crude antigen from C. sinensis recovered from the infected rabbits, the crude antigen from the adult worm excretory-secretory, and the crude antigen from billis of the rabbits resolved 26,27 and 19 profiles between 200-9 kDa, respectively. When E-64 supplemented 29, and 22 bands, respectively. More study should be carried out in the future on the immunological characteristics and the monoclonal antibody of the each antigen.