Differential diagnosis of Taenia asiatica infection from other human taeniases by serology has been tested. An enzyme-linked immunoelectrotransfer blot (EITB) was applied to subjected human sera and tapeworm materials. Thirty-eight proteins reactive to serum IgG were observed between 121 and 10 kDa in adult worms, and more than 22 serum-reactive components between 97 kDa and 21.5 kDa were observed in eggs of T. asiatica. Antigens of adult T. asiatica revealed immunoblot bands between 120 and 21.5 kDa against T. asiatica infected sera. Antigens of adult Taenia saginata revealed 110-100, 66, 58-56, and 46 kDa immunoblot bands against T. asiatica infected sera. Antigens of adult Taenia solium also revealed 99-97, 68-66, and 46 kDa bands against T. asiatica infected sera. The immunoblot band of 21.5 kDa exhibited specificity to T. asiatica.
Animals ; Antibodies, Helminth/immunology ; Antigens, Helminth/chemistry/immunology ; Humans ; Immunoblotting ; Molecular Weight ; Taenia/chemistry/*immunology ; Taeniasis/*immunology/parasitology

Excretory-secretory products of Toxocara canis larvae have been considered as a major functional antigen in immune responses against toxocariasis. We studied ultrastructural localization of T. canis second-stage larval antigen using a seropositive human serum under immunogold electron microscopy. High-density gold particles were observed in the secretory cells, excretory duct, intestinal epithelium, and cuticle of the larval worm sections. The distribution of the positive reactions in the larval worms suggests that the nature of the antigen is excretory-secretory antigen including waste metabolites and secretory enzymes.
Animals ; Antigens, Helminth/*immunology/ultrastructure ; Humans ; Larva/*immunology/ultrastructure ; Toxocara canis/*immunology/ultrastructure ; Toxocariasis/*immunology

This study was undertaken to characterize the properties of a 100 kDa somatic antigen from Metagonimus yokogawai. Monoclonal antibodies (mAbs) were produced against this 100 kDa antigen, and their immunoreactivity was assessed by western blot analysis with patients' sera. The mAbs against the 100 kDa antigen commonly reacted with various kinds of trematode antigens, including intestinal (Gymnophalloides seoi), lung (Paragonimus westermani), and liver flukes (Clonorchis sinensis and Fasciola hepatica). However, this mAb showed no cross-reactions with other helminth parasites, including nematodes and cestodes. To determine the topographic distribution of the 100 kDa antigen in worm sections, indirect immunoperoxidase staining was performed. A strong positive reaction was observed in the tegumental and subtegumental layers of adult M. yokogawai and C. sinensis. The results showed that the 100 kDa somatic protein of M. yokogawai is a common antigen which recognizes a target epitope present over the tegumental layer of different trematode species.
Animals ; Antibodies, Helminth/immunology ; Antibodies, Monoclonal/*immunology ; Antigens, Helminth/*immunology ; Clonorchis sinensis/immunology ; Cross Reactions/immunology ; Fasciola hepatica/immunology ; Female ; Helminth Proteins/*immunology ; Heterophyidae/*immunology ; Immunologic Tests ; Mice ; Mice, Inbred BALB C ; Paragonimus westermani/immunology ; Trematode Infections/*diagnosis/immunology

This study was performed to investigate whethere there is cross-reactivity between Dirofilaria immitis and three intestinal nematodes of dogs. In ELISA, D. immitis-infected dog sera obtained at the 4th molting stage (9-11 weeks) and microfilaremic stage (25-30 weeks) were shown to be highly reactive with crude extract of T. canis. In immunoblotting, some antigenic fractions, 44, 57, 88, 100 kDa of crude extract of T. canis, were found to be positive reaction with sera of dogs infected with D. immitis. However, little or no cross-reaction were observed between sera of D. immitis-infected dogs and crude extract antigen of T. vulpis or A. caninum. These result suggest that there are partial cross reaction between sera of D. immitis-infected dogs and the antigen of T. canis.
Animals ; Antibodies, Helminth/*immunology ; Antigens, Helminth/*immunology ; Cross Reactions ; Dirofilaria immitis/*immunology ; Dirofilariasis/*immunology ; Dogs ; Enzyme-Linked Immunosorbent Assay ; Immunoblotting ; Toxocara canis/*immunology

OBJECTIVE: To obtain the coding genes related to Schistosoma japonicum (Sj) cercariae 66 to approximately 68 kD antigens,and to provide antigens for diagnosis and vaccine of schistosomiasis. METHODS: Sj cercariae cDNA library was screened using the monospecific anti-sera of rabbit against soluble cercariae 66 to approximately 68 kD antigens as probes.The inserted cDNA fragments of the positive clones were amplified with PCR and identified by agarose gel electrophoresis. Four strong positive clones were further sequenced and analyzed through the internet NCBI/BLAST software. RESULTS: Twenty-one positive clones were obtained, 10 of which revealed a single band (0.5 to approximately 3.0 kb).The 4 strong positive clones showed high identity to SJCHGC05187,SJCHGC05173,SJCHGC06989, and SJCHGC01894 at the nucleotide level. CONCLUSION Four coding genes related with Sj antigens are obtained.
Animals ; Antibodies, Helminth ; immunology ; Antigens, Helminth ; immunology ; Cercaria ; genetics ; immunology ; DNA, Complementary ; genetics ; Gene Library ; Immune Sera ; immunology ; Schistosoma japonicum ; genetics ; immunology

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

Specific serum IgE levels of Clonorchis sinensis in infected humans were measured by avidin-biotin ELISA, and allergens from C. sinensis were identified by immunoblot and autoradiography. Then, allergens fractionated by Sephadex G-200 gel filtration were analyzed, and cross-reactive allergenic components of C. sinensis reacted with paragonimiasis sera were revealed. Fourteen out of 15 C. sinensis egg-positives were found to be serum IgE positive (absorbance > 0.27). Of 14 IgE-reacting allergen bands visualized, major allergens of 66, 61.5, 45, 37, 28.5, 23.5 and 15.5 KD were recognized by more than 50% of the sera of infected humans. The considerable individual variations of IgE immune responses to C. sinensis allergenic components were also noticed. C. sinensis extract was separated into 5 fractions by Sephadex G-200 gel filtration. Seventy-four KD allergen was recognized in the first fraction, 50, 45, 37, 29.5 and 28.5 KD in the third, and 15.5 KD in the fourth. Cross-reactive allergens with sera of paragonimiasis cases were identified as 66, 45, 28.5, 13 and 7.5 KD.
Allergens/*immunology ; Animal ; Antibodies, Helminth/*blood ; Antigens, Helminth/*immunology ; Clonorchiasis/*immunology ; Clonorchis sinensis/*immunology ; Cross Reactions ; Human ; Immunoglobulin E/*blood ; Support, Non-U.S. Gov't

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

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

OBJECTIVE: To determine the immune-protective effect of Japan Schistosoma (Chinese mainland strain) 23 kD membrane protein-heat shock protein (SjC23-Hsp70) DNA vaccine plus adjuvantinduced interleukin-12 (IL-12) plasmid DNA on Schistosoma japonicum infection in water buffalos. METHODS: Forty-five health water buffalos (8-10 months old) in non-endemic area of schistosomiasis were randomly assigned into group A (SjC23-Hsp70+IL-12, 300 μg), group B (SjC23+IL-12, 300 μg) and group C (pVAX+IL-12, 300 μg), 15 in each group. Each buffalo was immuned by shoulder intramuscular injection for 3 times, at an interval of 28 days. Twenty-eight days after the last immunization, each buffalo was infected with 1000 Japan cercariae of Schistosoma. Fecal examinations were conducted 2 days and 1 day before the perfusion, and on the day of perfusion. The number of hatching miracidia and eggs per gram feces was recorded. Fifty-six days after the infection, the buffalos were sacrificed and perfused via the descending aorta. The recovered adult worms and eggs in the liver tissue were counted. RESULTS: We compared group A and B with group C: the estrogen reduction rate was 45.7% and 26.61%; bug reduction rate was 44.51% and 25.84%; the fecal egg reduction rate was 41.1% and 31.63%; the miracidium reduction rate was 48.11% and 38.07%; and the liver egg reduction rate was 43.39% and 31.95%. The above rates in group A were higher than those in group B (P<0.05). CONCLUSION SjC23-Hsp70 DNA vaccine combined with IL-12 may have a significant immunoprotective effect on buffalos.
Animals ; Antigens, Helminth ; immunology ; Buffaloes ; Cattle ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Helminth Proteins ; immunology ; Immunization ; methods ; Interleukin-12 ; genetics ; immunology ; Membrane Proteins ; immunology ; Schistosomiasis japonica ; immunology ; prevention & control ; veterinary ; Vaccines, DNA ; administration & dosage ; immunology ; Vaccines, Synthetic ; immunology