ABSTRACT
A study was made to observe the association between the resistance to reinfection induced by irradiated metacercariae (MC) of Clonorchis sinensis and antigen specific Th1- and Th2-type cytokine productions in rats. Rats were infected with 20 MC of C. sinensis, previously exposed to a single dose of gamma irradiation, which varied from 0 to 100 Gy. All of them, single dose of 12 Gy showed higher IgG antibody titer with lowest worm recovery. Thus, 50 MC were used to challenge infection in rats previously infected with 20 MC irradiated at 12 Gy and the highest resistance to challenge infection was observed. The results of lymphocyte proliferation with specific antigen, ES Ag were shown no difference of proliferative responses as compared with primary and challenge infection at 12 Gy irradiation dose. In the case of cytokines production were observed that interferon (IFN-gamma) and interlukin (IL-2) were significantly enhanced, while IL-4 and IL-10 was almost unchanged to make comparison between primary and secondary infection at 12 Gy irradiation dose. In conclusion, the single dose of 12 Gy could be adopted for induction of the highest resistance to challenge infection. Up-regulation of Th1 type cytokines, IFN-gamma and IL-2 may be affected to develop vaccine by irradiated MC.
Subject(s)
Animals , Rats , Clonorchiasis/parasitology , Clonorchis sinensis/radiation effects , Cytokines/biosynthesis , Clonorchiasis/immunology , Clonorchis sinensis/pathogenicity , Gamma Rays , Immunoglobulin G/blood , Radiation Dosage , Th1 Cells/immunology , /immunologyABSTRACT
We investigated the induction of resistance to Clonorchis sinensis infection by prior infection in rat and hamster models. Animals were challenged with C. sinensis metacercariae, then treated with praziquantel and reinfected. Worm recovery rate in reinfected animals was used to estimate resistance to reinfection. The determined resistance rates to reinfection in rats and hamsters were 97.7% and 10.3%, respectively. In rats, cure from the primary infection of C. sinensis increased resistant to reinfection, and the greatert the worm burden and the longer the duration of primary infection, the higher was the resistance rate. For primary infection doses of 10, 40 and 100 metacercariae per rat, the resistance rates were 87.4%, 93.8% and 98.4%, respectively. The resistance rates in rats after 2 or 8-week primary infection were 78.7% and 95.3%, respectively. All worms recovered from reinfected rats were immature. When cured rats were administered with methylprednisolone, resistance to reinfection became impaired. These findings indicate that rats develop a high degree of resistance to reinfection by C. sinensis after cure. The growths and maturations of reinfected worms were also impaired.
Subject(s)
Animals , Female , Male , Rats , Anthelmintics/administration & dosage , Anti-Inflammatory Agents/administration & dosage , Clonorchiasis/immunology , Clonorchis sinensis/pathogenicity , Disease Models, Animal , Cricetinae , Immunocompetence , Immunosuppression Therapy , Mesocricetus , Methylprednisolone/administration & dosage , Praziquantel/administration & dosage , Rats, Sprague-DawleyABSTRACT
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 found 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 multicomplex protein originating from various organs of adult C. sinensis, and that it is composed of several 7 and 8 kDa proteins.
Subject(s)
Animals , Humans , Mice , Antigens, Helminth/immunology , Clonorchiasis/immunology , Clonorchis sinensis/anatomy & histology , Helminth Proteins/immunology , Mice, Inbred BALB C , Molecular WeightABSTRACT
Urine was concentrated 20-fold for assay for CAg of Schistosoma japonicum. mAb-RIHA and mAb-DotELISA were positive in 78, 31% and 65.06% of cases respectively, of 83 patients with acute schistosomiasis. The false positive rates in 101 healthy controls were 14.85% and 0%, respectively. Cross-reactions (using mAb-RIHA) were seen in 16.36% and 14.28% of patients with clonorchiasis, 49 patients with ankylostomiasis, respectively. Corresponding figures for mAb-DotELISA were 0% and 0%.
Subject(s)
Acute Disease , Antibodies, Monoclonal/diagnosis , Antigens, Helminth , Case-Control Studies , Clonorchiasis/immunology , Cross Reactions , Enzyme-Linked Immunosorbent Assay/methods , False Positive Reactions , Glycoproteins/immunology , Helminth Proteins/immunology , Hookworm Infections/immunology , Humans , Reproducibility of Results , Schistosomiasis japonica/drug therapy , Sensitivity and SpecificityABSTRACT
We report the use of immunogold-silver staining (IGSS), dot-ELISA and dot-IGSS methods in the study of clonorchiasis in China. These methods were employed to detect the antibody in sera from 40 clonorchiasis patients. The positive rates were 100%, 90.0% and 95.0%, respectively. When the three methods were used to examine 40 normal sera, the negative rates were 100%, 97.5% and 97.5%, respectively. These results suggest that IGSS, dot-ELISA and dot-IGSS are highly specific and sensitive in detecting anti-Clonorchis antibody in patients.
Subject(s)
Antibodies, Helminth/isolation & purification , Clonorchiasis/immunology , Enzyme-Linked Immunosorbent Assay/methods , Humans , Immunohistochemistry , Sensitivity and SpecificityABSTRACT
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.