This study was undertaken to evaluate the role of peritoneal exudate cells in the transfer of immunity against the liver fluke, Clonorchis sinensis in the inbred BALB/c mice. Ten donor mice were divided into 2 groups. One group consisted of 5 mice was infected orally with 20 metacercariae of C. sinensis, and the other group was injected intraperitoneally with 20 excysted larvae. Thirty days after immunization, the peritoneal exudate cells were obtained from the donor mice. Twenty recipient mice were divided into 4 equal groups for the purpose of primary immunization. The mice of Group I were injected intraperitoneally with 2 x 10(6) peritoneal exudate cells of the donor mice infected orally, those of Group III were injected intraperitoneally with 2 x 10(6) peritoneal exudate cells of the donor mice injected intraperitoneally. Those of Group II were injected orally with 20 metacercariae of C. sinensis. The group IV mice served as controls. Four days after the primary immunization all recipient mice were challenged orally with 20 metacercariae of C. sinensis, and then killed 30 days after the challenging infection. When the peritoneal exudate cells were injected into the recipient mice, pronounced reduction in eggs per gram of the feces was found in the mice of Group I and Group II, but no reduction in those of Group III. In the worm burdens of C. sinensis, the number of flukes found in the mice of Group II was only significantly less than those in the control group(IV). In addition the number of plaque forming cells per spleen in the mice of Group II was found larger than those in Group I. It is likely that donor peritoneal exudate cells transferred to the recipients might result in the production of relative immunity.
parasitology-helminth-trematoda ; Clonorchis sinensis ; immunology ; mouse

Thin layer immunoassay was carried out to demonstrate antibodies against Clonorchis sinensis in sera from clonorchiasis patients. Saline extract of adult worm was used as antigen. TIA technique was performed as described earlier by Elwing et al. (1976), but agarose was used instead of agar. The antibody titres of sera in 60 clonorchiasis cases were higher than that of 10 healthy and 10 amoebiasis cases, but not different comparing with that of 10 paragonimiasis cases. Antibody titres in clonorchiasis gave no differences according to the age, sex, EPG in feces, eosinophilia degree of blood, level of alkaline phosphatase and transaminase (SGOT, SGPT) in sera. It is suggested that, after evaluation, the TIA might supplement or be used as an alternative to other immunodiagnostic tests already in use for the diagnosis of clonorchiasis.
parasitology-helminth-trematoda ; Clonorchis sinensis ; clonorchiasis ; immunology

Thin layer immunoassay was carried out to demonstrate antibodies against Clonorchis sinensis in sera from clonorchiasis patients. Saline extract of adult worm was used as antigen. TIA technique was performed as described earlier by Elwing et al. (1976), but agarose was used instead of agar. The antibody titres of sera in 60 clonorchiasis cases were higher than that of 10 healthy and 10 amoebiasis cases, but not different comparing with that of 10 paragonimiasis cases. Antibody titres in clonorchiasis gave no differences according to the age, sex, EPG in feces, eosinophilia degree of blood, level of alkaline phosphatase and transaminase (SGOT, SGPT) in sera. It is suggested that, after evaluation, the TIA might supplement or be used as an alternative to other immunodiagnostic tests already in use for the diagnosis of clonorchiasis.
parasitology-helminth-trematoda ; Clonorchis sinensis ; clonorchiasis ; immunology

The sensitivity and specificity of crude and affinity-purified antigens of Clonorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent assay (ELISA) The results were as follows: The antibody-binding antigen (ABA) purified from whole worm crude antigen (WWA) by CNBr-activated Sepharose 4B affinity chromatography made 4 specific bands against rabbit anti-sera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16,300-18,500 and 28,000-29,000 daltons, while major ABA protein bands were at 18,000-21,000 and 29,000-31,000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA. Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15,000-200,000 daltons, 15,000-100,000 daltons and 11,000-80,000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31,000 daltons, while those of EGA and MEA consisted of higher molecular weights than 30,000 daltons. The ELISA reactivities of WWA to rabbit anti-sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4-6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting with WWA showed a plateau without variation. MEA showed relatively low O.D. values (<0.6), when reacted with anti-sera from lightly infected groups throughout the experiments, although there were some weak positive cases (O.D.>0.6) in heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion plates. Summarizing the above results, it is suggested that the whole worm antigen prepared from the adult worms of C. sinensis is most highly antigenic. However, this antigen might reveal cross reactions with other trematodes such as Paragonimus westermani, therefore, purification of antigenic proteins from the crude antigen is essential to increase the sensitivity and specificity for the immunodiagnosis of clonorchiasis.
parasitology-helminth-trematoda ; Clonorchis sinensis ; clonorchiasis ; rabbit ; immunology ; diagnosis ; antigen

The present study attempted to demonstrate an acquired immunity against Clonorchis sinensis in albino rat. Three separate experiments were carried out to determine acquired resistance developed by stimulating procedure followed by challenging infection with metacercariae. Acquired resistance was evaluated by the rate of recovery or the average number of flukes recovered from the liver of challenged albino rats, compared with the controls. In drder to demonstrate the rate of recovery of the fluke, three experimental groups of rats were challenged with 50 metacercariae per rat- 7,15 and 30 days after single injection of worm extract. The recovery rate was ranged from 33.2% to 38.0% in experimental group and their control group harbored from 37.8% to 42.6%. No significant difference was found on statistical analysis. In experimental groups received two immunizing injections with worm extract followed by challenging infection of metacercariae. Statistically significant difference was recovered between experimentals and controls. It was noted that reduction of the recovery rate was prominent in Group 5 and 6, which were challenged 15 and 30 days after two stimulating injections. From the third experiment which was consisted of single immunizing infection with 20 metacercariae followed by challenging infection with 30 metacercariae, no significant reduction was found between experimental rats and their controls. The number of recovered worms ranged from 16.2 to 18.5 worms in experimental group, while that of control group ranged from 18.9 to 19.8 worms. The evidence of delayed hypersensitivity reaction was observed in the groups with acquired immunity developed rats, by histopathological study of host hepatic tissue.
parasitology-helminth-trematoda ; Clonorchis sinensis ; immunology ; rat ; histology

: In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis (SDS-PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblotted. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable.
parasitology-helminth-trematoda ; Paragonimus westermani ; immunology ; antigen ; electrophoresis

In order to understand the effect of prednisolone injection on the immune responses of albino rat against Paragonimus iloktsuenensis infection, the differential leucocyte counts, appearance of immunoblast (large pyroninophilic cell, LPC) in the spleen and lungs in various experimental groups were observed in relation with the growth, maturation and migration sites of this rodent lungfluke. Rats of 3 experimental groups (A series), each group consisted of 5 rats, were infected with 20 metacercariae of P. iloktsuenensis, and they were kept for 3 days(Group I), 3 weeks(Group II), and 4 weeks (Group III) of infection period. The same number of experimental groups, each group of rats received 10 mg/kg dose of prednisolone injection every other day, were also kept and examined in comparison with the former groups. Preparation of peripheral blood smear and collection of worms were completed immediately after the end of infection period, and they were stained with Giemsa or Semichon's acetocarmine. Paraffin sections of the spleen and the lung tissues were stained with hematoxylin-eosin and methyl green pyronin (MGP). Those materials from A and B series of experimental groups were examined under the light microscope, and the results obtained were as follows: On observing differential leucocyte counts of peripheral blood smear, lymphocyte counts were consistently higher than those of uninfected controls in A series of infected groups, while those of B series were consistently low. On the other hand, neutrophil counts of A series showed lower counts than those of B series. In general, fluctuation patterns of both A and B series of experimental groups were almost the same, although lymphocyte and neutrophil counts showed reciprocal relation. The eosinophil counts of both series were negligible, especially in the groups of B series. The counts of LPC in the periarterial lymphatic sheath of the spleen were rapidly increased in the groups of A series, while those of B series were much less than those of A series, and the appearance of considerable LPC in the spleen was also delayed in B series. Furthermore, LPC of peribronchial lymphatic tissue in A series started to increase after the invasion of lungflukes into the lungs, while those of B series were much less due to the inhibited migration of lymphocytes into the lesions. Number, size and maturity of collected worms showed no significant differences between the groups of A and B series, but migration speed of the lungflukes was somewhat accelerated in B series than in A series. In this connection, it was considered that the immune responses of albino rats did not contribute for the complete protection against P. iloktsuenensis, but inhibited the migration of this lungfluke to some extent.
parasitology-helminth-trematoda ; Paragonimus iloktsuenensis ; paragonimiasis ; immunology ; prednisolone

parasitology-helminth-trematoda ; Paragonimus westermani ; paragonimiasis ; ELISA ; immunology

As epidemiological parameters of human paragonimiasis, the positive rates of intradermal test and the sputum/stool examination have long been employed in population surveys. However, both the specificity of the intradermal test and the sensitivity of sputum/stool examination have been gradually declined as the endemicity was lowered; thus the gap between above two parameters widened. In such context, the development of a new epidemiological parameter or tool which makes it possible to accurately discriminate the active paragonimiasis cases was necessary. In the present study, the detection rate of Paragonimus-specific IgG antibody by micro-ELISA was evaluated as an indicator of epidemiologic status of human paragonimiasis in a population. A total of 4,285 students and inhabitants living in Bukpyeong Myeon and Bukil Myeon, Haenam Gun, Jeonlanam Do was surveyed in October 1983 by intradermal test first. Out of them, 244 case (5.7 percent) were found positively reacted to VBS antigen of P. wetermani. Out of 168 positive reactors, 7 cases (4.2 percent) were egg positive either by two times of sputum examination or by one stool examination. That indicated that only 0.16 percent of total surveyed were confirmed as active paragonimiasis by egg detection. When sera collected from 239 positive reactors of intradermal test were tested by micro-ELISA for their specific IgG antibody, 40 cases (16.7 percent) were found to be positive. All of 7 eggs positive cases were again positive for specific IgG antibody. Among remaining 232 intradermal test positive cases, 33 cases were positive for IgG antibody. In contrast to those, none of 42 positive reactors to intradermal test for Clonorchis and of 128 intradermal test negative cases showed positive for Paragonimus- specific IgG antibody. The rate of specific IgG antibody as detected by micro-ELISA appeared to be increased with the wheal size of the intradermal test. When the wheal size was over 13 mm in diameter, about 50 percent of them were positive for specific IgG antibody. Thirty-one specific antibody positive cases were clinically evaluated by laboratory examinations (repeated sputum examination, peripheral eosinophil count and chest roentgenogram) and by history taking. Out of them 24 cases were associated with one or more positive laboratory findings; thus considered as active paragonimiasis cases. Out of 7 lab. finding-free cases 3 revealed past history of typical paragonimiasis symptoms, suggesting that they were in chronic or in convalescent stages. The remaining 4 cases were considered as either mild or ectopic infection cases; the possibility of cross-reaction with other helminthiases could not be ruled out. From the above results, it was inferred that the detection of Paragonimus-specific IgG antibogy by micro-ELISA was very much helpful in detecting the active cases as well as in proper evaluation of the endemicity of human paragonimiasis in a population. The convenience of mass handling of sera in micro-ELISA was considered another superiority as an epidemiologic tool.
parasitology-helminth-trematoda ; Paragonimus westermani ; paragonimiasis ; ELISA ; immunology ; diagnosis ; IgG

This experiment shows cellular and humoral immune responses induced by soluble egg antigen of Schistosoma mansoni, that is, change of the number of peripheral blood eosinophil, delayed hypersensitivity measured by the degree of ear swelling, granulomatous change of liver tissue and elevation of serum antibody titer by ELISA. SEA was given continuously by the insertion of a mini-pump into peritoneal cavity of mouse. In control group, same pump with HGG was inserted. New pump was exchanged once in two weeks and followed the result until 9 weeks after mini-pump insertion. Highest peripheral blood eosinophil level was recorded at 2-3 weeks after SEA pump insertion. Maximum ear swelling was observed at 2 weeks and then decreased gradually. In liver tissue, several granulomas without egg were formed at 4 weeks. Serum antibody titer was elevated from 4 weeks after SEA pump insertion.
parasitology-helminth-trematoda ; Schistosoma mansoni ; egg ; immunology ; mouse ; animal