Clonorchis sinensis is a liver fluke and it is the most prevalent human parasite in Korea at present. The parasite infection induces immune responses, characteristically an increased production of parasite-specific IgE in the host. Major IgE-reacting C. sinensis antigens in infected humans have been protein bands with MWs of 15, 28, 37, 45, 51, 56, 62, 66, 74, 97 and 160 KD identified by immunoblot analysis. Individual variations of the IgE binding pattern to C. sinensis antigens have also been documented. Using immune BALB/c mouse sera, IgE-reacting protein bands have been visualized with MWs of 28, 74, 86, 160 and several > 200 KD. One of the most strongly reacted C. sinensis antigenic proteins with a molecular weight of 28 KD was purified by gel filtration and preparative electrophoresis. Using a monoclonal antibody produced against the antigenic protein, the protein was localized in the parasite's intestine, and also found to be contained in excretory-secretory antigens.
Animal ; Antibodies, Monoclonal ; Antigens, Helminth/immunology* ; Antigens, Helminth/analysis* ; Clonorchis sinensis/immunology* ; Female ; Fluorescent Antibody Technique ; Human ; IgE/immunology* ; Immunoblotting ; Mice ; Mice, Inbred BALB C

OBJECTIVETo identify the egg antigens related to the formation of hepatic granulomas and fibrosis of Schistosomiasis japonica.

METHODSThe egg cDNA library of Schistosoma japonicum (S. japonicum) was constructed and screened by immunological methods with the pooled sera of advanced schistosomiasis patients. The inserted foreign DNA fragments of positive clones were sequenced. The sequence data were analyzed using Wdnasis 2.5 and compared with Genebank data using blast software.

RESULTSEighty-one clones containing recombinant DNA fragments were obtained from the egg cDNA library of S. japonicum by immunological screening. The DNA sequences of all clones belonged to the miracidial antigen family. The longest cDNA fragment was 1604 bp, which contained an open reading frame of 351 bp, which encoded a protein of 1 2913.35 daltons.

CONCLUSIONThe cDNA sequence of the miracidial antigen of S. japonicum (Chinese strain) was obtained for the first time.

Animals ; Antigens, Helminth ; genetics ; Base Sequence ; China ; DNA, Complementary ; analysis ; Ovum ; Schistosoma japonicum ; genetics ; immunology

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.
Animals ; Antigens, Helminth/*analysis ; Carrier Proteins/*immunology ; Cysticercosis/diagnosis/immunology ; Helminth Proteins/*immunology ; Humans ; Immunoblotting/methods ; Molecular Weight ; Serologic Tests ; Sparganum ; Taenia solium/*chemistry/immunology

We purified specific 31/36 kDa antigenic molecules from sparganum in different intermediate hosts (snakes and mice) and analyzed their monosaccharide compositions. Compositional analysis showed that glucose and mannose concentrations were 2-3 fold higher in the 31/36 kDa molecule purified from snakes than those from mice. This result implies that antigenic glycoproteins of sparganum from snakes might be modified in mammalian sparganosis with respect to their carbohydrate composition.
Animals ; Antigens, Helminth/*chemistry ; Human ; Mice ; Monosaccharides/*analysis ; Snakes/parasitology ; Sparganum/immunology ; Spirometra/*immunology ; Support, Non-U.S. Gov't

Although stool examination is the standard diagnostic method of clonorchiasis, serodiagnosis by ELISA using crude antigen is now widely used because of its convenience. However, ELISA diagnosis still suffers from cross-reactions, and therefore there is a need to improve the present conventional ELISA. The present study was undertaken to evaluate the diagnostic value of ELISA using excretory-secretory antigen (ESA) instead of crude antigen (CA) of Clonorchis sinensis. The diagnostic sensitivity of ELISA using excretory-secretory antigen was 92.5%, which was higher than that of ELISA using crude Clonorchis sinensis antigen (88.2%). In addition, the specificity of excretory-secretory antigen was found 93.1% while that of crude antigen was 87.8%. In summary, Clonorchis sinensis ESA was found to be a better serodiagnostic antigen than CA for ELISA.
Animals ; Antigens, Helminth/*analysis/immunology ; Clonorchiasis/*diagnosis/*immunology ; Clonorchis sinensis/immunology ; Enzyme-Linked Immunosorbent Assay/*methods ; Sensitivity and Specificity

An immunoelectron microscopy employing immunogold labeling method was performed to detect tissue origin of D1 fraction (D1A) among 5 antigenic protein fractions partially purified by DEAE-anion exchange chromatography from water-soluble crude antigen (PIWA) of adult Paragonimus iloktsuenensis. Immune reactions of adult worm tissues with rabbit serum immunoglobulin immunized with crude antigen (PI-Ig) and D1 antigen (D1-Ig), as well as rat serum immunoglobulin infected with P. iloktsuenensis were observed. D1A showed strong antigenicity in the intestinal epithelium of the worms during the early infection period of 2-4 weeks after infection. The vitellaria also showed stronger antigenicity than the other tissue sites in immune reaction of tissues against all immunoglobulins from 4 to 33 weeks after vitelline development. Therefore, it is suggested that D1A was mainly originated from the intestinal epithelial tissues before the development of vitelline gland of the parasites. Immuno-reactivity of two immunoglobulins (PI-Ig, D1-Ig) was significantly different in intestinal epithelial cytoplasmic protrusions (CP) and intestinal epithelial secretory granules (SG). In the experimental group with D1-Ig, gold particles were labeled significantly in CP than in SG when compared to the PI-Ig group. Thus, the major antigenic materials in D1 antigen having a strong antigenicity in the early infection period was considered to be originated from the intestinal epithelial tissue.
Animals ; Antigens, Helminth/*analysis/isolation & purification ; Chromatography, Ion Exchange ; Immunohistochemistry/methods ; Microscopy, Immunoelectron ; Paragonimus/*immunology ; Support, Non-U.S. Gov't

Three dogs were experimentally infected with Dirofilaria immitis. All dogs were euthanised at 30, 36 and 37 weeks after inoculation of D. immitis for the recovery of adult worms. Three cases accounted to 42.91 % recovery of inoculated worms. Serum samples from dogs experimentally inoculated with D. immitis were analyzed by ELISA and immunoblotting methods. Antibody titers of dogs detected by ELISA peaked between 7 and 14 weeks then decreased between weeks 15 to 24 followed by another increase during weeks 25 to 30 and persisted throughout the remainde of the experiment period. Analysis of adult D. immitis protein stained with Coomassie brilliant blue R-250 indicated separately more than 10 bands, and the major bands were 22, 40, 46, 56, 70, 72 and 89 kDa. Antigenic identification of extracts antigens of adults D. immitis by immunoblotting analysis revealed several bands from pooled sera of patent infection (30 weeks after inoculation). The detected bands were 24, 70, 80 and 110 kDa, 22, 72 and 84 kDa, and 58 and 72 kDa in dogs 1, 2 and 3, respectively. Results of antibody titers reached high levels on the 4th molting stage after inoculation of infective larva (L3), and reinforced previous findings that high molecular weight regions are detected in young animals.
Animals ; Antibodies, Helminth/analysis ; Antigens, Helminth/analysis/chemistry ; Autopsy ; Dirofilaria immitis/chemistry/*immunology ; Dirofilariasis/*immunology/parasitology ; Dog Diseases/*immunology/*parasitology ; Dogs/*immunology/*parasitology ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Female ; Immunoblotting ; Male ; Time Factors

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 organspecific 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.
Animals ; Antigens, Helminth/*analysis ; Clonorchis sinensis/anatomy & histology/*immunology ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Rabbits ; Research Support, Non-U.S. Gov't

The antigenic characterizations and serological reactions of human liver flukes, Clonorchis sinensis and Opisthorchis viverrini, were analyzed by immunoblot. The antigenic profiles of the crude extract of Clonorchis contained major proteins of 8, 26-28, 34-37, 43, and 70 kDa, and those of Opisthorchis 34-37, 43, 70, and 100 kDa. Of these, the 8, 26-28 and 34-37 kDa bands of Clonorchis and the 100 kDa of Opisthorchis were major components of each excretory-secretory antigen. The 8 and 26-28 kDa bands were specific to Clonorchis but the 100 kDa of Opisthorchis cross-reacted with the sera of clonorchiasis, and the 34-37, 70 and 100 kDa bands cross-reacted with sera of other helminthiases. The frequency and intensity of the immunoblot reactions were positively correlated with the intensity of the liver fluke infection.
Animals ; Antigens, Helminth/analysis/*chemistry/immunology ; Clonorchiasis/*diagnosis ; Clonorchis sinensis/*immunology ; Cross Reactions ; Electrophoresis, Polyacrylamide Gel/veterinary ; Humans ; Immunoblotting/veterinary ; Molecular Weight ; Opisthorchiasis/*diagnosis ; Opisthorchis/*immunology

In December 2011, we reported an autochthonous case of Echinococcus multilocularis infection in a 42-year-old woman in Korea. The diagnosis was based on histopathological findings of the surgically resected liver cyst. In the present study, we evaluated the serological and molecular characteristics of this Korean E. multilocularis case. The patient's serum strongly reacted with affinity-purified native Em18 and recombinant Em18 antigens (specific for E. multilocularis) but negative for recombinant antigen B8/1 (reactive for Echinococcus granulosus). In immunoaffinity chromatography, the serum also strongly reacted with E. multilocularis and only weakly positive for E. granulosus. We determined the whole nucleotide sequence of cox1 (1,608 bp) using the paraffin-embedded cystic tissue which was compared with E. multilocularis isolates from China, Japan, Kazakhstan, Austria, France, and Slovakia. The Korean case showed 99.8-99.9% similarity with isolates from Asia (the highest similarity with an isolate from Sichuan, China), whereas the similarity with European isolates ranged from 99.5 to 99.6%.
Adult ; Animals ; Antibodies, Helminth/*blood ; Antigens, Helminth/genetics/*immunology/metabolism ; Base Sequence ; Echinococcosis, Hepatic/*immunology/parasitology ; Echinococcosis, Pulmonary/diagnosis/genetics/immunology ; Echinococcus granulosus/genetics/immunology ; Echinococcus multilocularis/genetics/*immunology/isolation & purification ; Electron Transport Complex IV/genetics ; Female ; Humans ; Mitochondria/genetics ; Molecular Sequence Data ; Republic of Korea ; Sequence Analysis, DNA