Parasites, Hosts and Diseases

"Korean isolate"

Original Article

Immune Responses of Mice Intraduodenally Infected with Toxoplasma gondii KI-1 Tachyzoites: Eun-Hee Shin, Yeoun Sook Chun, Won-Hee Kim, Jae-Lip Kim, Kyoung-Ho Pyo, Jong-Yil Chai; Korean J Parasitol 2011;49(2):115-123.
Published online June 14, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.2.115

Abstract
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Toxoplasma gondii Korean isolate (KI-1) tachyzoites were inoculated intraduodenally to BALB/c mice using a silicon tube, and the course of infection and immune responses of mice were studied. Whereas control mice, that were infected intraperitoneally, died within day 7 post-infection (PI), the intraduodenally infected mice survived until day 9 PI (infection with 1×10⁵ tachyzoites) or day 11 PI (with 1×10⁶ tachyzoites). Based on histopathologic (Giemsa stain) and PCR (B1 gene) studies, it was suggested that tachyzoites, after entering the small intestine, invaded into endothelial cells, divided there, and propagated to other organs. PCR appeared to be more sensitive than histopathology to detect infected organs and tissues. The organisms spread over multiple organs by day 6 PI. However, proliferative responses of splenocytes and mesenteric lymph node (MLN) cells in response to con A or Toxoplasma lysate antigen decreased significantly, suggesting immunosuppression. Splenic CD4⁺ and CD8⁺ T-lymphocytes showed decreases in number until day 9 PI, whereas IFN-γ and IL-10 decreased slightly at day 6 PI and returned to normal levels by day 9 PI. No TNF-α was detected throughout the experimental period. The results showed that intraduodenal infection with KI-1 tachyzoites was successful but did not elicit significant mucosal immunity in mice and allowed dissemination of T. gondii organisms to systemic organs. The immunosuppression of mice included reduced lymphoproliferative responses to splenocytes and MLN cells to mitogen and low production of cytokines, such as IFN-γ, TNF-α, and IL-10, in response to T. gondii infection.

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Brief Communication

Suppression of CD4⁺ T-Cells in the Spleen of Mice Infected with Toxoplasma gondii KI-1 Tachyzoites: Won-Hee Kim, Eun-Hee Shin, Jae-Lip Kim, Seung-Young Yu, Bong-Kwang Jung, Jong-Yil Chai; Korean J Parasitol 2010;48(4):325-329.
Published online December 16, 2010; DOI: https://doi.org/10.3347/kjp.2010.48.4.325

Abstract
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Toxoplasma gondii KI-1, a recent new isolate from Korea, shows similar pathogenicity and infectivity to mice compared to the virulent RH strain. To understand characteristics of host immunity, including immune enhancement or suppression, we investigated proliferative responses and phenotypes of spleen cells. In addition, kinetics of IFN-γ, a Th1 cytokine, was examined in BALB/c mice up to day 6 post-infection (PI). Intraperitoneal injection of mice with 10³ KI-1 tachyzoites induced significant decreases (P < 0.05) in proliferative responses of spleen cells. This occurred at days 2-6 PI even when concanavalin A (con A) was added and when stimulated with KI-1 antigen, suggesting suppression of the immunity. CD4⁺ T-cells decreased markedly at day 2 PI (P < 0.05), whereas CD8⁺ T-cells, NK cells, and macrophages did not show significant changes, except a slight, but significant, increase of CD8⁺ T-cells at day 6 PI. The capacity of splenocytes to produce IFN-γ by con A stimulation dropped significantly at days 2-6 PI. These results demonstrate that intraperitoneal injection of KI-1 tachyzoites can induce immunosuppression during the early stage of infection, as revealed by the decrease of CD4⁺ T-cells and IFN-γ.

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Original Article

Proteomic Analysis of Toxoplasma gondii KI-1 Tachyzoites: Si-Hwan Choi, Tae Yun Kim, Sung Goo Park, Guang-Ho Cha, Dae-Whan Shin, Jong-Yil Chai, Young-Ha Lee; Korean J Parasitol 2010;48(3):195-201.
Published online September 16, 2010; DOI: https://doi.org/10.3347/kjp.2010.48.3.195

Abstract
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We studied on the proteomic characteristics of Toxoplasma gondii KI-1 tachyzoites which were originally isolated from a Korean patient, and compared with those of the well-known virulent RH strain using 2-dimensional electrophoresis (2-DE), mass spectrometry, and quantitative real-time PCR. Two-dimensional separation of the total proteins isolated from KI-1 tachyzoites revealed up to 150 spots, of which 121 were consistent with those of RH tachyzoites. Of the remaining 29 spots, 14 showed greater than 5-fold difference in density between the KI-1 and RH tachyzoites at a pH of 5.0-8.0. Among the 14 spots, 5 from the KI-1 isolate and 7 from the RH strain were identified using MALDI-TOF mass spectrometry and database searches. The spots from the KI-1 tachyzoites were dense granule proteins (GRA 2, 3, 6, and 7), hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGRPTase), and uracil phosphoribosyltransferase (UPRTase). The spots from the RH strain were surface antigen 1 (SAG 1), L-lactate dehydrogenase (LDH), actin, chorismate synthase, peroximal catalase, hexokinase, bifunctional dihydrofolate reductase-thymidylate synthase (DHTR-TS), and nucleoside-triphosphatases (NTPases). Quantitative real-time PCR supported our mass spectrometric results by showing the elevated expression of the genes encoding GRA 2, 3, and 6 and UPRTase in the KI-1 tachyzoites and those encoding GRA 7, SAG 1, NTPase, and chorismate synthase in the RH tachyzoites. These observations demonstrate that the protein compositions of KI-1 and RH tachyzoites are similar but differential protein expression is involved in virulence.

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Brief Communications

Genotyping of a Korean isolate of Toxoplasma gondii by multilocus PCR-RFLP and microsatellite analysis: Juan-Hua Quan, Tae Yun Kim, In-Uk Choi, Young-Ha Lee; Korean J Parasitol 2008;46(2):105-108.
Published online June 20, 2008; DOI: https://doi.org/10.3347/kjp.2008.46.2.105

Abstract
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Although the Korean isolate KI-1 of Toxoplasma gondii has been considered to be a virulent type I lineage because of its virulent clinical manifestations, its genotype is unclear. In the present study, genotyping of the KI-1 was performed by multilocus PCR-RFLP and microsatellite sequencing. For 9 genetic markers (c22-8, c29-2, L358, PK1, SAG2, SAG3, GRA6, BTUB, and Apico), the KI-1 and RH strains exhibited typical PCR-RFLP patterns identical to the type I strains. DNA sequencing of tandem repeats in 5 microsatellite markers (B17, B18, TUB2, W35, and TgM-A) of the KI-1 also revealed patterns characteristic of the type I. These results provide strong genetic evidence that KI-1 is a type I lineage of T. gondii.

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Evaluation of the Korean Isolate-1 Tachyzoite Antigen for Serodiagnosis of Toxoplasmosis: Eun-Hee Shin, Dong-Hee Kim, Aifen Lin, Jo-Woon-Yi Lee, Hyo-Jin Kim, Myoung-Hee Ahn, Jong-Yil Chai; Korean J Parasitol 2008;46(1):45-48.
Published online March 20, 2008; DOI: https://doi.org/10.3347/kjp.2008.46.1.45

Abstract
PDF

To evaluate the usefulness of the Korean Isolate-1 (KI-1) antigen for serodiagnosis of toxoplasmosis, antigen profiles of KI-1 tachyzoites were analyzed in comparison with RH tachyzoites by SDS-PAGE and immunoblotting. ELISA was performed on latex agglutination (LA)-positive and negative serum samples using KI-1 and RH antigens. Immunoblotting of the KI-1 antigen showed multiple antigen bands with molecular sizes of 22-105 kDa. Among them, 1 and 6 common bands were noted against a KI-1-infected and a RH-infected human serum, respectively, which represented differences in antigenic profiles between KI-1 and RH tachyzoites. However, all 9 LA-positive human sera were found positive by ELISA, and all 12 LA-negative sera were negative by ELISA; the correlation between the ELISA titers and LA titers was high (r = 0.749). Our results suggest that tachyzoites of KI-1 may be useful for serodiagnosis of human toxoplasmosis.

Citations

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Resistance toToxoplasma gondiiInfection in Mice Treated with Silk Protein by Enhanced Immune Responses
Joung-Ho Moon, Kyoung-Ho Pyo, Bong-Kwang Jung, Hyang Sook Chun, Jong-Yil Chai, Eun-Hee Shin
The Korean Journal of Parasitology.2011; 49(3): 303. CrossRef

8,175 View
64 Download
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Original Articles

Genetic characteristics of the Korean isolate KI-1 of Toxoplasma gondii: Aifen Lin, Eun-Hee Shin, Tae-Yun Kim, Jae-Hwan Park, Sang-Mee Guk, Jong-Yil Chai; Korean J Parasitol 2005;43(1):27-32.
Published online March 20, 2005; DOI: https://doi.org/10.3347/kjp.2005.43.1.27

Abstract
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Toxoplasma gondii tachyzoites were isolated from an ocular patient in the Republic of Korea and maintained in the laboratory (designated KI-1). In the present study, its genotype was determined by analyzing dense granule antigen 6 (GRA6) gene and surface antigen 2 (SAG2) gene as typing markers. Digestion of the amplification products of GRA6 and of the 5' and 3' ends of SAG2, respectively, with Mse I, Sau3A I, and Hha I, revealed that KI-1 is included in the genotype I, which includes the worldwide virulent RH strain. In addition, when the whole sequences of the coding regions of SAG1, rhoptry antigen 1 (ROP1), and GRA8 genes of KI-1 were compared with those of RH, minor nucleotide polymorphisms and amino acid substitutions were identified. These results show that KI-1 is a new geographical strain of T. gondii that can be included in the genotype I.

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Geographical distribution of Toxoplasma gondii genotypes in Asia: A link with neighboring continents
P. Chaichan, A. Mercier, L. Galal, A. Mahittikorn, F. Ariey, S. Morand, F. Boumédiène, R. Udonsom, A. Hamidovic, J.B. Murat, Y. Sukthana, M.L. Dardé
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The Korean Journal of Parasitology.2015; 53(3): 259. CrossRef
Genetic characterization of Toxoplasma gondii isolates from chickens in India by GRA6 gene sequence analysis
Shantaveer Biradar, Buddhi Saravanan, Anup Tewari, Chirukandoth Sreekumar, Muthu Sankar, Naduvanahalli Sudhakar
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Immune Responses of Mice Intraduodenally Infected withToxoplasma gondiiKI-1 Tachyzoites
Eun-Hee Shin, Yeoun Sook Chun, Won-Hee Kim, Jae-Lip Kim, Kyoung-Ho Pyo, Jong-Yil Chai
The Korean Journal of Parasitology.2011; 49(2): 115. CrossRef
Proteomic Analysis ofToxoplasma gondiiKI-1 Tachyzoites
Si-Hwan Choi, Tae Yun Kim, Sung Goo Park, Guang-Ho Cha, Dae-Whan Shin, Jong-Yil Chai, Young-Ha Lee
The Korean Journal of Parasitology.2010; 48(3): 195. CrossRef
Suppression of CD4+T-Cells in the Spleen of Mice Infected withToxoplasma gondiiKI-1 Tachyzoites
Won-Hee Kim, Eun-Hee Shin, Jae-Lip Kim, Seung-Young Yu, Bong-Kwang Jung, Jong-Yil Chai
The Korean Journal of Parasitology.2010; 48(4): 325. CrossRef
Selection of Polymorphic Peptides from GRA6 and GRA7 Sequences of Toxoplasma gondii Strains To Be Used in Serotyping
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Clinical and Vaccine Immunology.2009; 16(8): 1158. CrossRef
Genotyping of a Korean isolate of Toxoplasma gondii by multilocus PCR-RFLP and microsatellite analysis
Juan-Hua Quan, Tae Yun Kim, In-Uk Choi, Young-Ha Lee
The Korean Journal of Parasitology.2008; 46(2): 105. CrossRef
Evaluation of the Korean Isolate-1 Tachyzoite Antigen for Serodiagnosis of Toxoplasmosis
Eun-Hee Shin, Dong-Hee Kim, Aifen Lin, Jo-Woon-Yi Lee, Hyo-Jin Kim, Myoung-Hee Ahn, Jong-Yil Chai
The Korean Journal of Parasitology.2008; 46(1): 45. CrossRef
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Use of GRA6-Derived Synthetic Polymorphic Peptides in an Immunoenzymatic Assay To Serotype Toxoplasma gondii in Human Serum Samples Collected from Three Continents
Susana Sousa, Daniel Ajzenberg, Manuel Vilanova, José Costa, Marie-Laure Dardé
Clinical and Vaccine Immunology.2008; 15(9): 1380. CrossRef

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Laboratory passage and characterization of an isolate of Toxoplasma gondii from an ocular patient in Korea: Jong-Yil Chai, Aifen Lin, Eun-Hee Shin, Myoung-Don Oh, Eun-Taek Han, Ho-Woo Nam, Soon-Hyung Lee; Korean J Parasitol 2003;41(3):147-154.
Published online September 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.3.147

Abstract
PDF

Toxoplasma gondii tachyzoites were isolated from the blood of an ocular patient, and have been successfully passaged in the laboratory, for over a year, by peritoneal inoculation in mice. The isolated parasite was designated the Korean Isolate-1 (KI-1) and its characteristics were compared with those of the RH strain, a well-known virulent strain originating from a child who suffered from encephalitis. The morphology, pathogenicity, infectivity and cell culture characteristics of the KI-1 were similar to those of the RH strain. Both RH and KI-1 antigens were detected by an anti-T. gondii monoclonal antibody (mAb), Tg563, against the major surface protein SAG1 (30 kDa), whereas no reaction was observed against an anti-Neospora caninum mAb, 12B4. The KI-1 was confirmed as an isolate of T. gondii. A long-term laboratory maintenance and characterization of a local T. gondii isolate is reported for the first time in the Republic of Korea.

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Analysis of polymorphic region of GAM-1 gene in Plasmodium vivax Korean isolates: Weon-Gyu Kho, Joon-Yong Chung, Ui-Wook Hwang, Jin-Ho Chun, Yeong-Hong Park, Woo-Chul Chung; Korean J Parasitol 2001;39(4):313-318.
Published online December 31, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.4.313

Abstract
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The identification, characterization and quantification of Plasmodium sp. genetic polymorphism are becoming increasingly important in the vaccine development. We investigated polymorphism of Plasmodium vivax GAM-1 (PvGAM-1) gene in 30 Korean isolates. The polymorphic region of the PvGAM-1 gene, corresponding to nt 3792-4029, was amplified using polymerase chain reaction (PCR) followed by sequencing. All of the P. vivax Korean isolates were one type of GAM-1 gene, which were identical to that of the Belem strain. It is suggested that PvGAM-1 could not be used as a genetic marker for identifying or classifying P. vivax Korean isolates. It revealed that the polymorphic pattern was acquired basically by duplication and modification or deletion event of a 33 bp-motif fragment ended by poly guanine (G) and that there were at least three complete and one partial 33 bp-motif sequences within the polymorphic region in the longest cases such as those of South Korean and Belem isolates. In addition, we clustered P. vivax isolates with parsimonious criteria on the basis of PvGAM-1 polymorphic patterns (insertion/deletion patterns).

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Genetic Characteristics of Polymorphic Antigenic Markers among Korean Isolates of Plasmodium vivax
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Malaria Journal.2006;[Epub] CrossRef

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Analysis of polymorphic regions of Plasmodium vivax Duffy binding protein of Korean isolates: Weon-Gyu Kho, Joon-Yong Chung, Eun-Jeong Sim, Dong-Wook Kim, Woo-Chul Chung; Korean J Parasitol 2001;39(2):143-150.
Published online June 30, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.2.143

Abstract
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The present study was designed to investigate polymorphism in Duffy binding protein (DBP) gene of Plasmodium vivax isolates of Korea. Thirty samples were obtained from P. vivax patients in Yonchon-gun, Kyonggi-do in 1998. The PCR products of the samples were subjected to sequencing and hybridization analyses of the regions II and IV of P. vivax DBP gene. Two genotypes, SK-1 and SK-2, were identified on the basis of amino acid substitution and deletion. The genotype of 10 isolates was SK-1 and that of 20 isolates was SK-2. Most of the predicted amino acids in the region II of DBP gene were conserved between the Korean isolates and Belem strain except for 4-5 amino acid substitutions. In the region IV of DBP, a 6-bp insert that was shown in the Sal-1 allele type was found in SK-1, and a 27-bp insert that was shown in the Papua New Guinea allele type was found in SK-2. In conclusion, the present findings suggest that two genotypes of P. vivax coexist in the endemic area of Korea.

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Pathogenicity of Korean isolates of Acanthamoeba by observing the experimental infection and zymodemes of five isoenzymes: Kyung-Il Im, Ho-Joon Shin, Dong Whan Seo, Soung-Hoo Jeon, Tae-Eun Kim; Korean J Parasitol 1999;37(2):85-92.
Published online June 30, 1999; DOI: https://doi.org/10.3347/kjp.1999.37.2.85

Abstract
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To determine the pathogenicity of Acanthamoeba spp. isolated in Korea and to develop a isoenzymatic maker, the mortality rate of infected mice, in vitro cytotoxicity against target cells and isoenzyme band patterns were observed. Five isolates of Acanthamoeba spp. (YM-2, YM-3, YM-4, YM-5, and YM-7) were used in this study as well as three reference Acanthamoeba spp. (A. culbertsoni, A. hatchetti, and A. royreba). According to the mortality rate of infected mice, Korean isolates could be categorized into three groups: high virulent (YM-4), low virulent (YM-2, YM-5, YM-7) and the nonpathogenic group (YM-3). In addition, the virulence of Acanthamoeba spp. was enhanced by brain passage in mice. In the cytotoxicity assay against chinese hamster ovary cells, especially, the cytotoxicity of brain-passaged amoebae was relatively higher than the long-term cultivated ones. The zymodeme patterns of glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), hexokinase (HK), glutamate oxaloacetate transaminase (GOT) and malic enzyme (ME) of Acanthamoeba spp. were different among each isolate, and also between long-term cultured amoebae and brain passaged ones. In spites of the polymorphic zymodemes, a slow band of G6PD and HK, and an intermediate band of MDH were only observed in pathogenic Acanthamoeba spp., which should be used as isoenzymatic makers.

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Clinical Diagnostic Laboratory Immunology.2000; 7(3): 510. CrossRef