• Parasites, Hosts and Diseases
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• 제46권3호
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• pp.183-186
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• 2008

Helminthic cysteine proteases are well known to play critical roles in tissue invasion, nutrient uptake, and immune evasion of the parasites. In the same manner, the sparganum, the plerocercoid of Spirometra mansoni, is also known to secrete a large amount of cysteine proteases. However, cysteine protease inhibitors regulating the proteolytic activities of the cysteine protease are poorly illustrated. In this regard, we partially purified an endogenous cysteine protease inhibitor from spargana and characterized its biochemical properties. The cysteine protease inhibitor was purified by sequential chromatographies using Resource Q anion exchanger and Superdex 200 HR gel filtration from crude extracts of spargana. The molecular weight of the purified protein was estimated to be about 11 kD on SDS-PAGE. It was able to inhibit papain and 27 kDa cysteine protease of spargana with the ratio of 25.7% and 49.1%, respectively, while did not inhibit chymotrypsin. This finding suggests that the cysteine protease inhibitor of spargana may be involved in regulation of endogenous cysteine proteases of the parasite, rather than interact with cysteine proteases from their hosts.

• 대한수의학회지
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• 제13권1호
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• pp.75-77
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• 1973

A large plerocercoid was found from abdominal cavity of hen slaughtered in a slaughter-chicken house at Chyung Yang market in Seoul. The worm was identified as Diphyllobothrium mansoni of Genus Spirometra and the length of whole body was approximately 50 cm.

• Parasites, Hosts and Diseases
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• 제42권2호
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• pp.57-60
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• 2004

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

• Parasites, Hosts and Diseases
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• 제44권2호
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• pp.167-169
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• 2006

After infection of male mice with the plerocercoids (spargana) of Spirometra mansoni, serum levels of estrogen and testicular weight were analyzed by enzyme-linked immunosorbent assay (ELISA) and weighing machine, respectively. The serum level of estrogen increased progressively in infected mice compared with normal controls, whereas the testicular weight of infected mice decreased significantly (P < 0.05). These results suggest that certain substances from spargana change the steroid hormone metabolisms in the host by unknown pathways, and chronic infection may contribute to change of the function of steroid hormone target organ, i.e., testis, in male mice.

• Parasites, Hosts and Diseases
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• 제47권1호
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• pp.69-71
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• 2009

The tegument of tapeworms is known to be composed of an outer syncytial cytoplasm layer which includes microtriches and cytoplasmic organelles (= syncytial layer), and a parenchymatous cytoplasm layer that contains subtegumental cell nuclei (= subtegumental layer) and organelles. In the present study, separation of the syncytial layer of the sparganum, the plerocercoid stage of Spirometra mansoni, was tried using urea as the chemical reagent. Histological sections were prepared to visualize the status of separation after staining with hematoxylin and eosin. The results showed that the syncytial layer of the sparganum tegument which includes microtriches and cytoplasmic organelles were successfully separated from the parenchyma using 3 M urea.

• Parasites, Hosts and Diseases
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• 제48권2호
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• pp.183-185
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• 2010

In a previous study, the author developed a method for separation of the tegument of spargana (plerocercoids of Spirometra mansoni) from the parenchyme using urea. The present study, as a next step, was performed to evaluate which molecules are present in the outer tegument. Two major proteins, 180 and 200 kDa, are present in the tegument and we could make polyclonal antibodies against these molecules. Their immunolocalization was processed and the outermost layer of the spargana showed strong positive staining. Conclusively, we could confirm that the 180 and 200 kDa molecules might be tightly bound membrane proteins in the tegument of spargana.

• Parasites, Hosts and Diseases
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• 제42권3호
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• pp.141-143
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• 2004

After collecting calcareous corpuscles from plerocercoid of Spirometra mansoni (sparganum), we evaluated the antigenic values of calcareous corpuscles binding proteins obtained from the cyst fluid of Taenia solium metacestodes. Immunoblot analysis revealed that cysticercosis patient sera strongly recognized 10 and 95 kDa calcareous corpuscles binding proteins. This result demonstrated that calcareous corpuscles are bound with major secretory antigenic proteins, which is possibly involved in the secretory pathways of the 10 and 95 kDa proteins presenting in the cyst fluid of T. solium metacestodes.

• Parasites, Hosts and Diseases
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• 제30권3호
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• pp.227-230
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• 1992

스파르가눔증의 항체검사에서 항원으로 사용하는 스파르가눔 충체추출액의 구성 단백질 중, 환자혈청과 반응 시켜 관찰한 바, 분자량 36 및 31 kDa인 단백질이 항원성이 높았다. 또한 이 두가지 단백질은 스파르가눔 충체의 표피와 표피세포에 분포하여 충체 외부로 분비되는 항원일 것으로 추정하였다. 이 연구단보에서는 이들 두가지 단백질이 분비배설항원에 나타남을 증명하고자 스파르가눔을 $4^{\circ}C{\;}및{\;}37^{\circ}C$ 생리식염수에 30분~100시간 동안 배양하여 분비배설항원을 만들고, 분비배설항원내 단백질 분비량, 단백질의 조성 미치 단백질분해효소의 활성을 각각 관찰하였다. 충체 g당 분비배설항원의 단백질량은 관찰 범위안에서 배양온도에 관계없이 평균 7.7 mg이었다. SDS-폴리아크릴아미드 젤 전기영동상 분비배설항원의 단백질 구성은 분자량 66 kDa 이상인 단백질에서 차이가 있을 뿐 충체추출액의 것과 거의 유사하였다. 물론 분자량 36 및 31 kDa인 단백질도 분비배설항원의 주 구성성분이었다. 분비배설항원의 단백질분해효소 비활성도(比活性度)는 2.9~5.3 units/mg으로 충체추출액의 것과 차이가 없었다. 이미 보고된 스파르가눔의 cysteine protease가 체외로 분비되는 효소라는 것이 사실이라고 추정하게 하는 결과로 생각하였다.

• Parasites, Hosts and Diseases
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• 제29권1호
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• pp.1-8
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• 1991

스파르가눔 생리식염수 추출액 내에 포함되어 있는 성분단백질 중 스파르가눔증 환자 혈청내 특이 IgG항체와 민감하고 특이하게 반응하는 항원단벼질인 36, 29 kDa단백질을 단세포군 항체를 이용한 면역친화성 크로마토그 래피로 순수분리할 수 있음은 이미 보고하였다. 이 연구에서는 스파르가눔 추출액 내에 포함된 이 36, 29 kDa단백질이 젤라틴을 고리로 한 친화성 크로마토그래피로 훨씬 쉽게 순수분리할 수 있음을 증명하고자 하였다. 젤라틴을 고리로 부착시킨 Sepharose 4B column에 스파르가눔 추출액을 통과시키고 젤라틴에 부착한 단백질은 4 M urea/0.1M NaCl 용액을 분리완충액으로 분리하였다. 이렇게 분리한 단백질은 SDS-PAGE에서 36, 29 kDa band로 구성되어 있었고, SDS-PAGE/immunoblot 결과 환자의 polyclonal 항체는 이들 band에만 반응하였다. 스파르가눔증, 기타 기생충증 환자 및 건강대조군 혈청내 스파르가눔 특이항체가(IgG)를 면역효소측정 법으로 측정 한 결과 순수분리한 이 단백질은 특히 특이도가 95.8%로 생리식염수 추출액의 89%보다 우수하였고 민감도는 차이가 없었다. 이상의 결과는 젤라틴을 고리로 이용한 친화성 크로마토그래피는 스파르가눔 생리식염수 추출액 내의 36 및 29 kDa 단백질을 간편하게 순수분리할 수 있고 단백질의 항원성도 유지할 수 있음을 보이고 있었다.

• Parasites, Hosts and Diseases
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• 제38권3호
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• pp.145-150
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• 2000

Antigenic components in the crude extracts of Spirometra mansoni plerocercoid were analyzed in early experimental infections and in IgG subclass observed in clinical sparganosis. By IgG immunoblot, sera obtained serially from experimental mice, fed 5 spargana each, were reacted with the crude extracts. Protein bands at 36-26 kDa and 103 kDa showed positive reactions since two weeks after infection. In a differential immunoblot, in which a monospecific antibody against sparganum chymase at 36 kDa was pre-treated, the reactions at 36-26 kDa disappeared, indicating that the sparganum chymase and its degradation products invoked IgG antibody reactions. When 69 patients sera of human sparganosis were examined for their IgG subclass responses, IgG4 levels showed the highest reaction which was followed by IgG 1 The IgG4 antibody also reacted mainly with 36-31 kDa protease. These results indicate that 36 kDa chymase of 5. nansoni plerocercoid is the main antigenic component inducing Ige antibody response in early stage of experimental sparganosis and for specific IgG subclass reactions in human sparganosis.