1.Immunoblot Patterns of Taenia asiatica Taeniasis.
Hyeong Kyu JEON ; Keeseon S EOM
The Korean Journal of Parasitology 2009;47(1):73-77
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
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Antibodies, Helminth/immunology
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Antigens, Helminth/chemistry/immunology
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Humans
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Immunoblotting
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Molecular Weight
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Taenia/chemistry/*immunology
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Taeniasis/*immunology/parasitology
2.Serum Antigen and Antibody Detection in Echinococcosis: Application in Serodiagnosis of Human Hydatidosis.
Seyed Mahmoud SADJJADI ; Farzaneh SEDAGHAT ; Seyed Vahid HOSSEINI ; Bahador SARKARI
The Korean Journal of Parasitology 2009;47(2):153-157
Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.
Adult
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Antibodies, Helminth/*blood
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Antigens, Helminth/*blood
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Echinococcosis/*diagnosis
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Enzyme-Linked Immunosorbent Assay
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Female
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Humans
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Male
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Sensitivity and Specificity
3.Metagonimus yokogawai: a 100-kDa Somatic Antigen Commonly Reacting with Other Trematodes.
Eun Taek HAN ; Hyun Jong YANG ; Young Jin PARK ; Jeong Hyun PARK ; Jong Yil CHAI
The Korean Journal of Parasitology 2014;52(2):201-204
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
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Antibodies, Helminth/immunology
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Antibodies, Monoclonal/*immunology
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Antigens, Helminth/*immunology
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Clonorchis sinensis/immunology
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Cross Reactions/immunology
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Fasciola hepatica/immunology
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Female
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Helminth Proteins/*immunology
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Heterophyidae/*immunology
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Immunologic Tests
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Mice
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Mice, Inbred BALB C
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Paragonimus westermani/immunology
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Trematode Infections/*diagnosis/immunology
4.Nanoparticles as a vaccine adjuvant of anti-idiotypic antibody against schistosomiasis.
Zhen-qing FENG ; Shi-gen ZHONG ; Yu-hua LI ; Yun-qian LI ; Zhen-ning QIU ; Zhu-ming WANG ; Jun LI ; Li DONG ; Xiao-hong GUAN
Chinese Medical Journal 2004;117(1):83-87
BACKGROUNDThe development of new adjuvants for human use has been the focus of attention. This study's aim is to explore the possibility of using nanoparticle Ca nanoparticles (CA) as a vaccine adjuvant of anti-idiotypic antibody NP30 against schistosomiasis and its protective mechanisms.
METHODSNanoparticle CA-NP30 conjugate (CA-NP30) was fabricated. BALB/c mice were immunized actively with CA-NP30 to evaluate its effects of protective immunity on mice. The serum levels of specific IgG, IgG1 and IgG2a antibodies against NP30 and the concentrations of IFN-gamma and IL-4 in supernatant of splenocytes were determined via ELISA.
RESULTSNanoparticle CA could enhance significantly the protective immunity of NP30 against infection of Schistosoma japonicum and the worm reduction rose from 36.0% (NP30 alone) to 52.6%. The serum levels of specific IgG, IgG1 and IgG2a antibodies against NP30 increased remarkably, as compared with those of the group immunized with NP30 alone. The concentration of IFN-gamma in supernatant of splenocyte was drastically elevated [the groups immunized with CA-NP30 and NP30 alone were (493.80 +/- 400.74) pg/ml and (39.03 +/- 39.58) pg/ml, respectively], but the concentration of IL-4 showed no significant difference from that of NP30 alone [(27.94 +/- 9.84) pg/ml vs (27.28 +/- 14.44) pg/ml].
CONCLUSIONSNanoparticle CA could act as a vaccine adjuvant of anti-idiotypic antibody NP30 against schistosomiasis. The mechanism could be that CA-NP30 enhances humoral and cellular immune responses in mice.
Adjuvants, Immunologic ; Animals ; Antibodies, Anti-Idiotypic ; immunology ; Antibodies, Helminth ; immunology ; Mice ; Mice, Inbred BALB C ; Nanotechnology ; Schistosomiasis ; prevention & control ; Vaccines
5.Spatial distribution characteristics of the prevalence of advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody in Hunan Province in 2020.
Y ZHOU ; L TANG ; Y TONG ; J HUANG ; J WANG ; Y ZHANG ; H JIANG ; N XU ; Y GONG ; J YIN ; Q JIANG ; J ZHOU ; Y ZHOU
Chinese Journal of Schistosomiasis Control 2023;35(5):444-450
OBJECTIVE:
To investigate the spatial distribution characteristics of the prevalence of advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody, and to examine the correlation between the prevalence of advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody in Hunan Province in 2020, so as to provide insights into advanced schistosomiais control in the province.
METHODS:
The epidemiological data of schistosomiasis in Hunan Province in 2020 were collected, including number of permanent residents in survey villages, number of advanced schistosomiasis patients, number of residents receiving serological tests and number of residents seropositive for anti-Schistosoma antibody, and the prevalence advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody were descriptively analyzed. Village-based spatial distribution characteristics of prevalence advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody were identified in Hunan Province in 2020, and the correlation between the revalence advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody was examined using Spearman correlation analysis.
RESULTS:
The prevalence of advanced schistosomiasis was 0 to 2.72% and the seroprevalence of anti-Schistosoma antibody was 0 to 20.25% in 1 153 schistosomiasis-endemic villages in Hunan Province in 2020. Spatial clusters were identified in both the prevalence of advanced schistosomiasis (global Moran's I = 0.416, P < 0.01) and the seroprevalence of anti-Schistosoma antibody (global Moran's I = 0.711, P < 0.01) in Hunan Province. Local spatial autocorrelation analysis identified 98 schistosomiasis-endemic villages with high-high clusters of the prevalence of advanced schistosomiasis, 134 endemic villages with high-high clusters of the seroprevalence of anti-Schistosoma antibody and 36 endemic villages with high-high clusters of both the prevalence of advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody in Hunan Province. In addition, spearman correlation analysis showed a positive correlation between the prevalence of advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody (rs = 0.235, P < 0.05).
CONCLUSIONS
There were spatial clusters of the prevalence of advanced schistosomiasis and seroprevalence of anti-Schistosoma antibody in Hunan Province in 2020, which were predominantly located in areas neighboring the Dongting Lake. These clusters should be given a high priority in the schistosomiasis control programs.
Animals
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Humans
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Prevalence
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Seroepidemiologic Studies
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Schistosomiasis/epidemiology*
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Schistosoma
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Spatial Analysis
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Antibodies, Helminth
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China/epidemiology*
6.Detection of specific serum IgE in clonorchiasis cases and analysis of Clonorchis sinensis allergens.
Tai Soon YONG ; Dong Soo KIM ; Soo Young LEE ; Kyung Il IM ; Ki Young LEE
Yonsei Medical Journal 1993;34(3):248-257
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
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Animal
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Antibodies, Helminth/*blood
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Antigens, Helminth/*immunology
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Clonorchiasis/*immunology
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Clonorchis sinensis/*immunology
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Cross Reactions
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Human
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Immunoglobulin E/*blood
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Support, Non-U.S. Gov't
7.Cross-reactivity between sera from dogs experimentally infected with Dirofilaria immitis and crude extract of Toxocara canis.
Kun Ho SONG ; Mineo HAYASAKI ; Kyu Woan CHO ; Sang Eun LEE ; Duck Hwan KIM
The Korean Journal of Parasitology 2002;40(4):195-198
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
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Antibodies, Helminth/*immunology
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Antigens, Helminth/*immunology
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Cross Reactions
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Dirofilaria immitis/*immunology
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Dirofilariasis/*immunology
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Dogs
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Enzyme-Linked Immunosorbent Assay
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Immunoblotting
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Toxocara canis/*immunology
8.Identification of IgE-reacting Clonorchis sinensis antigens.
Tai Soon YONG ; Soon Jung PARK ; Du Ho LEE ; Hye Jin YANG ; Jongweon LEE
Yonsei Medical Journal 1999;40(2):178-183
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
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Antibodies, Monoclonal
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Antigens, Helminth/immunology*
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Antigens, Helminth/analysis*
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Clonorchis sinensis/immunology*
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Female
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Fluorescent Antibody Technique
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Human
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IgE/immunology*
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Immunoblotting
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Mice
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Mice, Inbred BALB C
9.Expression profiling and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum in the liver of infected New Zealand white rabbits.
Dan XIA ; Ganming DENG ; Pingying TENG ; Yu XIE ; Yaomin LI ; Chunmei WANG ; Shujie CHEN ; Minfang CHEN ; Rongjia MAI ; Haiyan LIAO ; Lingyu SHI ; Liyan OU ; Qiwei CHEN ; Xiaoguang CHEN ; Xiaohong ZHOU
Journal of Southern Medical University 2015;35(6):826-831
OBJECTIVETo examine the expression profile and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum (Sjp40) during granuloma formation in the liver of infected New Zealand white rabbits.
METHODSNew Zealand white rabbits were infected with S. japonicum cercariae, and the livers were harvested at 29 and 45 days post-infection (dpi). The total RNA of the liver tissues was extracted for expression profiling of Sjp40 by quantitative reverse transcription-PCR (qRT-PCR) with GAPDH of S. japonicum as the endogenous reference gene. The expression of Sjp40 in the liver were detected by Western blotting using anti-Sjp40 monoclonal antibody (mAb) 9G7 or anti-Toxoplasma gondii tSAG1 mAb Y3A8 (control) as the primary antibody. Paraffin sections of the liver were prepared for observing egg granuloma formation using HE staining and for indirect immunofluorescence assay of Sjp40 location in the trapped eggs and egg granulomas.
RESULTSThe level of Sjp40 mRNA in the eggs trapped in rabbit livers was significantly higher at 45 dpi than that at 29 dpi (P<0.05), and Western blotting confirmed the presence of Sjp40 protein in the rabbit livers at both 29 and 45 dpi. Immunofluorescence assay demonstrated localized expression of Sjp40 in the immature eggs in the rabbit liver at 29 dpi, but at 45 dpi fluorescence was detected in clusters of mature eggs containing miracidium and in the surrounding egg granulomas.
CONCLUSIONSThe transcriptional levels of Sjp40 significantly increased with the maturation of eggs trapped in the rabbit livers. Sjp40 protein spread from the eggs to the surrounding egg granuloma at 45 dpi when acute liver granulomatous lesions occur, suggesting that Sjp40 plays a key role in egg granulomas formation in the livers of infected New Zealand white rabbits.
Animals ; Antibodies, Monoclonal ; Antigens, Helminth ; metabolism ; Fluorescent Antibody Technique ; Gene Expression Profiling ; Granuloma ; parasitology ; Helminth Proteins ; metabolism ; Liver ; parasitology ; RNA, Messenger ; Rabbits ; Schistosoma japonicum ; Schistosomiasis japonica
10.Immuno-screening of Schistosoma japonicum cercariae cDNA library by the sera of anti-soluble cercariae 66 to approximately 68 kD antigens.
Yong-Hua QIN ; Shuai-Feng ZHOU ; Shi-Ping WANG
Journal of Central South University(Medical Sciences) 2008;33(12):1076-1081
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
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Antibodies, Helminth
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immunology
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Antigens, Helminth
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immunology
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Cercaria
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genetics
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immunology
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DNA, Complementary
;
genetics
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Gene Library
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Immune Sera
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immunology
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Schistosoma japonicum
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genetics
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immunology