• Restorative Dentistry and Endodontics
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• 제19권2호
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• pp.485-498
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• 1994

Porphyromonas endodontalis is a black-pigmented anaerobic Gram-negative rod which is associated with endodontal infections and this microorganism possesses a potential for pathogenicity. The purpose of this study was to compare the membrane components of Porphyromonas endodontalis and Porphyromonas gingivalis and to study the immune reaction patterns of Porphyromonas endodontalis with patients with periapical lesion. Porphyromonas endodontalis (ATCC 35406), Porphyromonas gingivals serotypea (381), serotype b(W50), serotype c(A7A1-28) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells and human sera were obtained from patients and dental students. Indirect immunofluorescence method was used to study on the cross reaction between Porphyromonas endodontalis and Porphyromonas gingivalis serotype a, b, c antigen. Total membrane protein profiles of Porphyromonas endodontalis antigen were studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and the reactivity of antigenic components of Porphyromonas endodontalis against sera of patients and rabbit anti-Porphyromonas endodontalis antisera were assessed by Immunoblotting method. The following results were obtained : 1. Antigens of Porphyromonas endodontalis has multiple antigenic components, and both patients with periapical lesion and normal healthy individual showed immune response to this. 2. Patients group and healthy individual group showed a diversity of immune reaction pattern but they showed immune response against 43kd protein. 3. Patients with periapical lesion showed more diverse immune response than healthy individual and in some patients, much more bands appeared to lower molecular weight protein. 4. According to indirect immunofluorescence and Immunoblotting study, Porphyromonas endodontalis did not share common antigen with Porphyromonas gingivalis serotype a, b, c.

• Parasites, Hosts and Diseases
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• 제40권2호
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• pp.83-88
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• 2002

The 8 kDa antigenic protein of Clonorchis sinensis was partially purified by ammonium sulfate precipitation and subsequently by a column chromatographic steps. The purified protein was separated into 7 and 8 kDa protein bands through SDS-tricine gel electrophoresis, while the protein was fecund to migrate to a 8 kDa band in 7.5-15% SDS-PAGE. The molecular weight of the antigen was estimated to be 110 kDa by Superose 6 HR 10/30 gel filtration. The purified antigen strongly reacted with the human sera of clonorchiasis. The hyperimmune sera of BALB/c mice immunized against the 8 kDa protein were reacted with both the crude extract and the excretory-secretory product of adult worms, but not with the metacercarial extract. Immunohistochemical staining demonstrated that the protein was distributed to the tegument and subtegumental cells and also to the seminal receptacle. The present findings suggest that the 8 kDa protein is a partition of the multicomplek protein originating from various organs of adult C. sinenis, and that it is composed of several 7 and 8 kDa proteins.

• Parasites, Hosts and Diseases
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• 제42권4호
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• pp.169-174
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• 2004

This study was carried out to find out specific proteins from different organs of Clonorchis sinensis. Crude extract, organ-specific and excretory-secretory (ES) proteins were analyzed by immunoblot with infected human sera. The bands of 7- and 17 -kDa were main component of intestinal fluid and ES protein and commonly found in all organ-specific proteins. The 17-kDa protein was observed from ES antigen, intestinal fluid, eggs and sperms, 26- and 28-kDa proteins were from the uterus, vitellaria, and ovary, and 34-, 37-, 43- and 50-kDa proteins were mainly from the testis and sperms. Serum of mice immunized with sperms reacted to the 50-kDa protein by immunoblotting and immunohistochemical staining showed a positive reaction at the seminal receptacle and seminiferous tubule. The present results show that the 7-kDa protein is a common antigen of every part or organ of C. sinensis, but different organs express their specific antigenic protein bands.

• Parasites, Hosts and Diseases
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• 제46권1호
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• pp.45-48
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• 2008

To evaluate the usefulness of the Korean Isolate-1 (KI-1) antigen for serodiagnosis of toxoplasmosis, antigen profiles of KI-1 tachyzoites were analyzed in comparison with RH tachyzoites by SOS-PAGE and immunoblotting. ELISA was performed on latex agglutination (LA)-positive and negative serum samples using KI-1 and RH antigens. Immunoblotting of the KI-1 antigen showed multiple antigen bands with molecular sizes of 22-105 kDa. Among them, 1 and 6 common bands were noted against a KI-1-infected and a RH-infected human serum, respectively, which represented differences in antigenic profiles between KI-1 and RH tachyzoites. However, all 9 LA-positive human sera were found positive by ELISA, and all 12 LA-negative sera were negative by ELISA; the correlation between the ELISA titers and LA titers was high (r = 0.749). Our results suggest that tachyzoites of KI-1 may be useful for serodiagnosis of human toxoplasmosis.

• 한국식품과학회지
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• 제42권5호
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• pp.627-631
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• 2010

국내에서 소비되는 콩 14품종에 대한 항원성 검사를 토끼에서 생산된 다클론 항체와 콩 알레르기 환자 혈청을 이용하여 검사하였다. 그 결과 콩 특이성을 갖는 다클론 항체를 이용한 연구에서는 일부 콩 품종에서 항원성을 나타내는 단백질이 추가적으로 존재하거나 결여된 것을 확인할 수 있었으며, 14품종 중 특히 진품콩에서는 다른 품종보다 항원성이 강한 단백질이 확인되었다. CAP검사에 의해 콩 항원에 대한 특이 항체 값이 65 U/mL 이상인 콩 알레르기 환자 4명의 혈청을 이용한 반응성 비교에서는 다른 품종에 비해 단백콩과 신팔달2호에서 조사된 모든 콩 알레르기 환자혈청과 상대적으로 뚜렷이 높은 반응성을 나타냈고, 대원콩에서 제일 낮은 반응성을 나타냈다. 이러한 결과는 콩 품종에 따라 포함된 단백질의 알레르기성 차이가 있음을 의미하며 콩을 원료로 식품을 제조할 때 콩 품종을 고려함으로써 알레르기 반응으로 인한 문제를 줄일 수 있는 가능성을 제시한다.

• Parasites, Hosts and Diseases
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• 제33권3호
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• pp.187-194
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• 1995

정상 흰쥐의 출생 후 연령과 면역억제된 흰쥐의 감염강도에 따른 혈청 내 폐포자충(Pneumocystis carinii)에 대한 항체 형성을 관찰하고자 이 연구를 수행하였다. 폐포자충을 발현시키고 순수분리하여 수용성 항원을 만들고 이를 이용하여 전기영동과 면역이적법(Immunoblot)과 효소면역법(ELISA)을 시행하여 항체반응을 관찰하였다. SDS-PAGE에 의하여 분리된 항원의 단백 질 분획은 20-200 kDa의 범위에서 20개 이상이 관찰되었다. 이들 분획 중에서 인체 양성표준 혈청에 116 kDa 분획이 강하게, 45-55 kDa 및 100 kDa 분획이 약하게 반응하였고, 흰쥐 양성표준 혈청과는 40-45 kDa 분회과 강하게 그리고 50-55 kDa, 116, 200 kDa 분획과 약하게 반응하였다. 연령 별로 이 네 분획과의 반응을 보면 출생 5일-6주까지 양성률 50-100%이고 8주에는 0% 가 되었고, 10주 이후에는 증가하여 40주에 100%가 되었다. 폐포자충의 감염량이 증가하여 감염 강도가 IV인 흰쥐에서는 면역이적으로 측정할만큼 항체를 가진 개체가 없었다. 이러한 정상과 면역 억제된 흰쥐의 혈청반응 유형은 효소면역법에서도 확인할 수 있었다. 네 항원 분획 중 40-45 kDa에 대한 혈청반응이 각 군에서 낮게 관찰되었으나 그 생물학적 의의는 아직 평가하기 어렵다. 이 결과는 흰쥐가 출생하면 모체에서 유래한 혈청 내 항폐포자충 항체를 8주가 경과하면서 모두 소실하고 그 이후에 10주부터 40주에 이르기까지 자연감염에 의해 항체를 서서히 형성하는 것을 의미한다. 면역 억제에 의하여 흰쥐가 항체를 효과적으로 생산하지 못하게 되면 혈청 내 항체량에 반비례하여 폐포자충의 감염량이 늘어난다. 이는 항체에 의해 매개되는 체액면역이 폐포자충의 감염과 직접 관련이 있다는 증거가 된다.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.129-136
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• 1997

Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

• Parasites, Hosts and Diseases
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• 제51권3호
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• pp.313-317
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• 2013

According to increase of travel, the cases of imported echinococcosis have been increasing in Korea. The present study was undertaken to develop a serodiagnostic system for echinococcosis in Korea. For diagnosis of echinococcosis, the fluid of Echinococcus granulosus hydatid cysts was collected from naturally infected sheep in Uzbekistan. Also serum samples of infected patients who were surgically confirmed were collected in a hospital in Tashkent, Uzbekistan. According to the absorbance of 59 echinococcosis positive and 39 negative control serum samples, the cut-off value was determined as 0.27. The sensitivity and specificity of ELISA with hydatid fluid antigen were 91.5% and 96%, respectively. The antigen cross-reacted with the serum of some cysticercosis or clonorchiasis patients. However, immunoblot analysis on the cystic fluid recognized antigenic proteins of 7-, 16-, and 24-kDa bands in their dominant protein quantity and strong blotting reactivity. In conclusion, the present ELISA system using hydatid cyst fluid antigen from Uzbekistan sheep is sensitive and specific for diagnosis of echinococcosis cases.

• Parasites, Hosts and Diseases
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• 제46권4호
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• pp.279-283
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• 2008

To examine humoral immune responses in the host, we measured serum antibody levels in different strains of mice (ICR, BALB/c, and C3H) experimentally infected with Neodiplostomum seoulense. Specific IgG antibody levels were increased remarkably with little difference among 3 strains of mice infected with N. seoulense from day 7 to 35 post-infection. More target proteins of adult parasites reacted with IgG at the time when the worm recovery decreased compared with other times. More than 20 protein bands, from 14 kDa to 94 kDa in size, were separated from the crude antigen of N. seoulense adults by SDS-PAGE, and among them 26, 30, 35, 43, 54, 67, and 94 kDa proteins were the major antigenic proteins. The results suggest that significant IgG antibody responses occur against N. seoulense in mice and this may be related with expulsion of worms.

• 대한수의학회지
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• 제30권2호
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• pp.223-229
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• 1990

A splenectomized 5-month-old calf was inoculated with cryopreserved Theileria sergenti infected blood originated from naturally infected Korean native cattle in Chonbuk district. At peak parasitemia (40.1%), blood was collected, washed, lysed and then the T sergenti merozoite was isolated by differential centrifugation. Antigenic profile of isolated T sergenti organism was analized by SDS-PAGE and western blotting techniques. Coomassie blue stained SDS-PAGE gel revealed at least twelve protein bands of approximately 14Kd, 28Kd, 30Kd, 34Kd, 36Kd, 38Kd, 41Kd, 56Kd, 66Kd, 72Kd, 97Kd and 116Kd in the merozoite homogenate. In western blot, although T sergenti antigen recognized by specific anti-T sergenti antibodies demonstrated 28Kd, 30Kd, 38Kd, 56Kd, 58Kd, 66Kd, 97Kd and 116Kd proteins. False positive reactions were also observed in normal bovine serum with T sergenti and normal erythrocytic antigens. Therefore, predominant proteins of T sergenti merozoite antigen were found to be 28Kd, 30Kd, and 41Kd proteins of molecular weights. On going studies we will analyze the relative importance of those antigens for immunity of T sergenti in Korean native cattle.