Parasites, Hosts and Diseases

"RT-PCR"

Original Articles

Production of IL-1β and Inflammasome with Up-Regulated Expressions of NOD-Like Receptor Related Genes in Toxoplasma gondii-Infected THP-1 Macrophages: Jia-Qi Chu, Ge Shi, Yi-Ming Fan, In-Wook Choi, Guang-Ho Cha, Yu Zhou, Young-Ha Lee, Juan-Hua Quan; Korean J Parasitol 2016;54(6):711-717.
Published online December 31, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.6.711

Abstract
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Toxoplasma gondii is an obligate intracellular parasite that stimulates production of high levels of proinflammatory cytokines, which are important for innate immunity. NLRs, i.e., nucleotide-binding oligomerization domain (NOD)-like receptors, play a crucial role as innate immune sensors and form multiprotein complexes called inflammasomes, which mediate caspase-1-dependent processing of pro-IL-1β. To elucidate the role of inflammasome components in T. gondii-infected THP-1 macrophages, we examined inflammasome-related gene expression and mechanisms of inflammasome-regulated cytokine IL-1β secretion. The results revealed a significant upregulation of IL-1β after T. gondii infection. T. gondii infection also upregulated the expression of inflammasome sensors, including NLRP1, NLRP3, NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and NAIP, in a time-dependent manner. The infection also upregulated inflammasome adaptor protein ASC and caspase-1 mRNA levels. From this study, we newly found that T. gondii infection regulates NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and neuronal apoptosis inhibitor protein (NAIP) gene expressions in THP-1 macrophages and that the role of the inflammasome-related genes may be critical for mediating the innate immune responses to T. gondii infection.

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Investigating the possible role of toxoplasmosis and Interleukin-1β variants on the immune response in Egyptian diabetic patients
Asmaa Ibrahim, Nancy O. kamel, Fatma Rageh, Rasha Elgamal, Mohamed A. Sakr, Eman M. Osman, Samar s. Ahmed, Hend A. Yassin, Yasmine N. Kamel, Reham F. Othman, Manar Ezzelarab Ramadan
Microbes and Infection.2025; 27(7): 105559. CrossRef
Early responses of primary human and bovine monocytes, monocytic THP-1 cells and THP-1 cell-derived macrophages to vital Toxoplasma gondii tachyzoites
Dominik Hanke, Zahady D. Velásquez, Kathrin Büttner, Andreas Krueger, Ralf Ross, Andreas Hecker, Sybille Mazurek, Veronika Grau, Anja Taubert, Carlos Hermosilla, Katrin Richter, Iván Conejeros
Frontiers in Immunology.2025;[Epub] CrossRef
Unraveling the NLRP family: Structure, function, activation, critical influence on tumor progression, and potential as targets for cancer therapy
Xueqing Zhou, Yongguang Tao, Ying Shi
Cancer Letters.2024; 605: 217283. CrossRef
Role of inflammasomes in Toxoplasma and Plasmodium infections
Zhi-xin Wang, Wan-jun Jiao, Yong Yang, Hong-li Liu, Hai-long Wang
Parasites & Vectors.2024;[Epub] CrossRef
NLRP13 inflammasome complex is hypermethylated in familial Mediterranean fever and global methylation correlates with the disease severity
Feyzanur Yildirimtepe Caldiran, Koksal Deveci, Ercan Cacan
Annals of Human Genetics.2023; 87(3): 115. CrossRef
Overview of Apoptosis, Autophagy, and Inflammatory Processes in Toxoplasma gondii Infected Cells
Ehsan Ahmadpour, Farhad Babaie, Tohid Kazemi, Sirous Mehrani Moghaddam, Ata Moghimi, Ramin Hosseinzadeh, Veeranoot Nissapatorn, Abdol Sattar Pagheh
Pathogens.2023; 12(2): 253. CrossRef
P2X7 Receptor Modulation of the Gut Microbiota and the Inflammasome Determines the Severity of Toxoplasma gondii-Induced Ileitis
Aline Cristina Abreu Moreira-Souza, Hayandra Ferreira Nanini, Thuany Prado Rangel, Sthefani Rodrigues Batista da Silva, Beatriz Pêgo Damasceno, Beatriz Elias Ribeiro, Cynthia M. Cascabulho, Fabiano Thompson, Camille Leal, Patrícia Teixeira Santana, Siane
Biomedicines.2023; 11(2): 555. CrossRef
The role of NOD-like receptors in innate immunity
Cássio Luiz Coutinho Almeida-da-Silva, Luiz Eduardo Baggio Savio, Robson Coutinho-Silva, David M. Ojcius
Frontiers in Immunology.2023;[Epub] CrossRef
NLRP3 Plays a Key Role in Antihelminth Immunity in the Enteral and Parenteral Stages of Trichinella spiralis-Infected Mice
Tian-Xu Pan, Hai-Bin Huang, Hui-Nan Lu, Guang-Xun Zhao, Yu Quan, Jun-Yi Li, Ying Xue, Zhi-Yu Zhu, Yue Wang, Chun-Wei Shi, Nan Wang, Gui-Lian Yang, Chun-Feng Wang, De’Broski R. Herbert
Infection and Immunity.2023;[Epub] CrossRef
Toxoplasma gondii profilin induces NLRP3 activation and IL-1β production/secretion in THP-1 cells
Hossein Pazoki, Hamed Mirjalali, Maryam Niyyati, Seyed Javad Seyed Tabaei, Nariman Mosaffa, Shabnam Shahrokh, Hamid Asadzadeh Ahdaei, Andreas Kupz, Mohammad Reza Zali
Microbial Pathogenesis.2023; 180: 106120. CrossRef
Possible therapeutic targets for NLRP3 inflammasome-induced breast cancer
Xixi Wang, Junyi Lin, Zhe Wang, Zhi Li, Minghua Wang
Discover Oncology.2023;[Epub] CrossRef
Toxoplasma gondii Induces Pyroptosis in Human Placental Trophoblast and Amniotic Cells by Inducing ROS Production and Activation of Cathepsin B and NLRP1/NLRP3/NLRC4/AIM2 Inflammasome
Juan-Hua Quan, Fei Fei Gao, Tian-Zhong Ma, Wei Ye, Xiang Gao, Ming-Zhu Deng, Lan-Lan Yin, In-Wook Choi, Jae-Min Yuk, Guang-Ho Cha, Young-Ha Lee, Jia-Qi Chu
The American Journal of Pathology.2023; 193(12): 2047. CrossRef
Role of Microgliosis and NLRP3 Inflammasome in Parkinson’s Disease Pathogenesis and Therapy
Fillipe M. de Araújo, Lorena Cuenca-Bermejo, Emiliano Fernández-Villalba, Silvia L. Costa, Victor Diogenes A. Silva, Maria Trinidad Herrero
Cellular and Molecular Neurobiology.2022; 42(5): 1283. CrossRef
Latent Upregulation of Nlrp3, Nlrc4 and Aim2 Differentiates between Asymptomatic and Symptomatic Trichomonas vaginalis Infection
Sonal Yadav, Vivek Verma, Rakesh Singh Dhanda, Sumeeta Khurana, Manisha Yadav
Immunological Investigations.2022; 51(5): 1127. CrossRef
P. aeruginosa biofilm activates the NLRP3 inflammasomes in vitro
Qi Tan, Qing Ai, Yu He, Fang Li, Jialin Yu
Microbial Pathogenesis.2022; 164: 105379. CrossRef
In silico analysis of expression and DNA methylation profiles of NLRP13 inflammasome in tumor cells
Feyzanur Yildirimtepe Caldiran, Caglar Berkel, Koksal Deveci, Ercan Cacan
Human Gene.2022; 33: 201067. CrossRef
The strategies of NLRP3 inflammasome to combat Toxoplasma gondii
Chanjin Yoon, Yu Seong Ham, Woo Jin Gil, Chul-Su Yang
Frontiers in Immunology.2022;[Epub] CrossRef
Expression profiles of NOD-like receptors and regulation of NLRP3 inflammasome activation in Toxoplasma gondii-infected human small intestinal epithelial cells
Jia-Qi Chu, Fei Fei Gao, Weiyun Wu, Chunchao Li, Zhaobin Pan, Jinhui Sun, Hao Wang, Cong Huang, Sang Hyuk Lee, Juan-Hua Quan, Young-Ha Lee
Parasites & Vectors.2021;[Epub] CrossRef
RIPK3 Facilitates Host Resistance to Oral Toxoplasma gondii Infection
Patrick W. Cervantes, Bruno Martorelli Di Genova, Billy Joel Erazo Flores, Laura J. Knoll, Jeroen P. J. Saeij
Infection and Immunity.2021;[Epub] CrossRef
Soluble total antigen derived from Toxoplasma gondii tachyzoites increased the expression levels of NLRP1, NLRP3, NLRC4, AIM2, and the release of mature form of IL1β, but downregulated the expression of IL1β and IL18 genes in THP-1cell line
Hossein Pazoki, Hanieh Mohammad Rahimi, Hamed Mirjalali, Maryam Niyyati, Nariman Mosaffa, Seyed Javad Seyed Tabaei, Shabnam Shahrokh, Hamid Asadzadeh Aghdaei, Mohammad Reza Zali
Microbial Pathogenesis.2021; 158: 105072. CrossRef
Host inflammatory responses to intracellular invaders: Review study
Ramesh Chandra Rai
Life Sciences.2020; 240: 117084. CrossRef
Treatment of mice with S4B6 IL-2 complex prevents lethal toxoplasmosis via IL-12- and IL-18-dependent interferon-gamma production by non-CD4 immune cells
Andreas Kupz, Saparna Pai, Paul R. Giacomin, Jennifer A. Whan, Robert A. Walker, Pierre-Mehdi Hammoudi, Nicholas C. Smith, Catherine M. Miller
Scientific Reports.2020;[Epub] CrossRef
Neospora caninum infection induces an isolate virulence-dependent pro-inflammatory gene expression profile in bovine monocyte-derived macrophages
Marta García-Sánchez, Laura Jiménez-Pelayo, Pilar Horcajo, Esther Collantes-Fernández, Luis Miguel Ortega-Mora, Javier Regidor-Cerrillo
Parasites & Vectors.2020;[Epub] CrossRef
A Comparison of Transcriptional Diversity of Swine Macrophages Infected With TgHB1 Strain of Toxoplasma gondii Isolated in China
Yongle Song, Lindong Song, Xiaoting Wan, Bang Shen, Rui Fang, Min Hu, Junlong Zhao, Yanqin Zhou
Frontiers in Cellular and Infection Microbiology.2020;[Epub] CrossRef
Cytosolic Recognition of Microbes and Pathogens: Inflammasomes in Action
Jenni A. Hayward, Anukriti Mathur, Chinh Ngo, Si Ming Man
Microbiology and Molecular Biology Reviews.2018;[Epub] CrossRef
P2X7 receptor mediates NLRP3-dependent IL-1β secretion and parasite proliferation in Toxoplasma gondii-infected human small intestinal epithelial cells
Juan-Hua Quan, Rui Huang, Zhuang Wang, Shuai Huang, In-Wook Choi, Yu Zhou, Young-Ha Lee, Jia-Qi Chu
Parasites & Vectors.2018;[Epub] CrossRef

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Overexpression of Ubiquitin and Amino Acid Permease Genes in Association with Antimony Resistance in Leishmania tropica Field Isolates: Elham Kazemi-Rad, Mehdi Mohebali, Mohammad Bagher Khadem-Erfan, Homa Hajjaran, Ramtin Hadighi, Ali Khamesipour, Sassan Rezaie, Mojtaba Saffari, Reza Raoofian, Mansour Heidari; Korean J Parasitol 2013;51(4):413-419.
Published online August 30, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.4.413

Abstract
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The mainstay therapy against leishmaniasis is still pentavalent antimonial drugs; however, the rate of antimony resistance is increasing in endemic regions such as Iran. Understanding the molecular basis of resistance to antimonials could be helpful to improve treatment strategies. This study aimed to recognize genes involved in antimony resistance of Leishmania tropica field isolates. Sensitive and resistant L. tropica parasites were isolated from anthroponotic cutaneous leishmaniasis patients and drug susceptibility of parasites to meglumine antimoniate (Glucantime®) was confirmed using in vitro assay. Then, complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) and real-time reverse transcriptase-PCR (RT-PCR) approaches were utilized on mRNAs from resistant and sensitive L. tropica isolates. We identified 2 known genes, ubiquitin implicated in protein degradation and amino acid permease (AAP3) involved in arginine uptake. Also, we identified 1 gene encoding hypothetical protein. Real-time RT-PCR revealed a significant upregulation of ubiquitin (2.54-fold), and AAP3 (2.86-fold) (P<0.05) in a resistant isolate compared to a sensitive one. Our results suggest that overexpression of ubiquitin and AAP3 could potentially implicated in natural antimony resistance.

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Brief Communication

Quantitative Evaluation of Viability- and Apoptosis-Related Genes in Ascaris suum Eggs under Different Culture-Temperature Conditions: Yong-Man Yu, You-Hang Cho, Young-Nam Youn, Juan Hua Quan, In-Wook Choi, Young-Ha Lee; Korean J Parasitol 2012;50(3):243-247.
Published online August 13, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.3.243

Abstract
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Ascaris suum eggs are inactivated by composting conditions; however, it is difficult to find functional changes in heat-treated A. suum eggs. Here, unembryonated A. suum eggs were incubated at 20℃, 50℃, and 70℃ in vitro, and the gene expression levels related to viability, such as eukaryotic translation initiation factor 4E (IF4E), phosphofructokinase 1 (PFK1), and thioredoxin 1 (TRX1), and to apoptosis, such as apoptosis-inducing factor 1 (AIF1) and cell death protein 6 (CDP6), were evaluated by real-time quantitative RT-PCR. No prominent morphological alterations were noted in the eggs at 20℃ until day 10. In contrast, the eggs developed rapidly, and embryonated eggs and hatched larvae began to die, starting on day 2 at 50℃ and day 1 at 70℃. At 20℃, IF4E, PFK1, and TRX1 mRNA expression was significantly increased from days 2-4; however, AIF1 and CDP6 mRNA expression was not changed significantly. IF4E, PFK1, and TRX1 mRNA expression was markedly decreased from day 2 at 50℃ and 70℃, whereas AIF1 and CDP6 mRNA expression was significantly increased. The expressions of HSP70 and HSP90 were detected for 9-10 days at 20℃, for 3-5 days at 50℃, and for 2 days at 70℃. Taken together, incremental heat increases were associated with the rapid development of A. suum eggs, decreased expression of genes related to viability, and earlier expression of apoptosis-related genes, and finally these changes of viability- and apoptosis-related genes of A. suum eggs were associated with survival of the eggs under temperature stress.

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Original Articles

Real-Time RT-PCR on SAG1 and BAG1 Gene Expression during Stage Conversion in Immunosuppressed Mice Infected with Toxoplasma gondii Tehran Strain: Monavar Selseleh, Mohammad Hossein Modarressi, Mehdi Mohebali, Saeedeh Shojaee, Mohammad Reza Eshragian, Mina Selseleh, Ebrahim Azizi, Hossein Keshavarz; Korean J Parasitol 2012;50(3):199-205.
Published online August 13, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.3.199

Abstract
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Toxoplasmic encephalitis is caused by reactivation of bradyzoites to rapidly dividing tachyzoites of the apicomplexan parasite Toxoplasma gondii in immunocompromised hosts. Diagnosis of this life-threatening disease is problematic, because it is difficult to discriminate between these 2 stages. Toxoplasma PCR assays using gDNA as a template have been unable to discriminate between an increase or decrease in SAG1 and BAG1 expression between the active tachyzoite stage and the latent bradyzoite stage. In the present study, real-time RT-PCR assay was used to detect the expression of bradyzoite (BAG1)- and tachyzoite-specific genes (SAG1) during bradyzoite/tachyzoite stage conversion in mice infected with T. gondii Tehran strain after dexamethasone sodium phosphate (DXM) administration. The conversion reaction was observed in the lungs and brain tissues of experimental mice, indicated by SAG1 expression at day 6 after DXM administration, and continued until day 14. Bradyzoites were also detected in both organs throughout the study; however, it decreased at day 14 significantly. It is suggested that during the reactivation period, bradyzoites not only escape from the cysts and reinvade neighboring cells as tachyzoites, but also converted to new bradyzoites. In summary, the real-time RT-PCR assay provided a reliable, fast, and quantitative way of detecting T. gondii reactivation in an animal model. Thus, this method may be useful for diagnosing stage conversion in clinical specimens of immunocompromised patients (HIV or transplant patients) for early identification of tachyzoite-bradyzoite stage conversion.

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ANK1 and DnaK-TPR, Two Tetratricopeptide Repeat-Containing Proteins Primarily Expressed in Toxoplasma Bradyzoites, Do Not Contribute to Bradyzoite Differentiation
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Brain cystogenesis capacity of Toxoplasma gondii, avirulent Tehran strain in mice
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Gene Expression Profiles in Genetically Different Mice Infected with Toxoplasma gondii: ALDH1A2, BEX2, EGR2, CCL3 and PLAU: Hassan Ahmed Hassan Ahmed Ismail, Juan-Hua Quan, Zhou Wei, In-Wook Choi, Guang-Ho Cha, Dae-Whan Shin, Young-Ha Lee, Chang-June Song; Korean J Parasitol 2012;50(1):7-13.
Published online March 6, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.1.7

Abstract
PDF

Toxoplasma gondii can modulate host cell gene expression; however, determining gene expression levels in intermediate hosts after T. gondii infection is not known much. We selected 5 genes (ALDH1A2, BEX2, CCL3, EGR2 and PLAU) and compared the mRNA expression levels in the spleen, liver, lung and small intestine of genetically different mice infected with T. gondii. ALDH1A2 mRNA expressions of both mouse strains were markedly increased at day 1-4 postinfection (PI) and then decreased, and its expressions in the spleen and lung were significantly higher in C57BL/6 mice than those of BALB/c mice. BEX2 and CCR3 mRNA expressions of both mouse strains were significantly increased from day 7 PI and peaked at day 15-30 PI (P<0.05), especially high in the spleen liver or small intestine of C57BL/6 mice. EGR2 and PLAU mRNA expressions of both mouse strains were significantly increased after infection, especially high in the spleen and liver. However, their expression patterns were varied depending on the tissue and mouse strain. Taken together, T. gondii-susceptible C57BL/6 mice expressed higher levels of these 5 genes than did T. gondii-resistant BALB/c mice, particularly in the spleen and liver. And ALDH1A2 and PLAU expressions were increased acutely, whereas BEX2, CCL3 and EGR2 expressions were increased lately. Thus, these demonstrate that host genetic factors exert a strong impact on the expression of these 5 genes and their expression patterns were varied depending on the gene or tissue.

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Changes of cytokine mRNA expression and IgG responses in rats infected with Capillaria hepatica: Dong-Kwan Kim, Kyoung-Hwan Joo, Myung-Sook Chung; Korean J Parasitol 2007;45(2):95-102.
Published online June 20, 2007; DOI: https://doi.org/10.3347/kjp.2007.45.2.95

Abstract
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The mRNA expression of several cytokines was evaluated in splenocytes and mesenteric lymph node (MLN) cells of rats infected with Capillaria hepatica by reverse-transcription (RT)-PCR until week 12 after infection. IgG1 and IgG2a, which are associated with Th1 and Th2 response, respectively, were also assessed by ELISA. The results indicated that the majority of cytokines, including the Th1 (IL-2 and IFN-γ) and Th2 cytokines (IL-4, IL-5 and IL-10) were expressed at maximal levels during the early stage of infection (after week 1-2), and the ELISA data also evidenced a similar pattern of changes in IgG1 and IgG2a. Th1 and Th2 cytokines responded in a similar fashion in this rat model. The expression of cytokines in splenocytes was significantly higher than that in MLN cells, thereby indicating that cytokine production is controlled more by spleen than by MLN. In addition, the observation that IFN-γ expression increased unexpectedly at the time of maximal egg production (6 weeks after infection) indicated that IFN-γ is a cytokine reacting against egg production. However, increased IL-5 expression occurring in tandem with worm activity indicated that the activity of C. hepatica might be controlled by IL-5 expression.

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Identification of differentially expressed cDNAs in Acanthamoeba culbertsoni after mouse brain passage: Kyu-Lee Han, Jongweon Lee, Don-Soo Kim, Soon-Jung Park, Kyung-il Im, Tai-Soon Yong; Korean J Parasitol 2006;44(1):15-20.
Published online March 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.1.15

Abstract
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Free-living amoebae of the genus Acanthamoeba are causative agents of granulomatous amebic encephalitis and amebic keratitis. Because the virulence of Acanthamoeba culbertsoni cultured in the laboratory is restored by consecutive brain passages, we examined the genes induced in mouse brain-passaged A. culbertsoni by differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Enhanced A. culbertsoni virulence was observed during the second mouse brain passage, i.e., infected mouse mortality increased from 5% to 70%. Ten cDNAs induced during mouse brain passage were identified by DDRT-PCR and this was confirmed by northern blot analysis. BlastX searches of these cDNAs indicated the upregulations of genes encoding predictive NADH-dehydrogenase, proteasomal ATPase, and GDP-mannose pyrophosphorylase B, which have previously been reported to be associated with A. culbertsoni virulence factors.

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Brief Communication

Changes of splenocyte IFN-γ mRNA synthesis in rats infected with Paragonimus westermani: Jun Kyong Cho, Hye Soo Kwon, Kyoung Hwan Joo, Joon Sang Lee, Sung-Weon Cho; Korean J Parasitol 1999;37(4):285-287.
Published online December 31, 1999; DOI: https://doi.org/10.3347/kjp.1999.37.4.285

Abstract
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Changes in the expression level of splenocyte IFN-γ mRNA of Sprague-Dawley (SD) rats infected with Paragonimus westermani were analyzed by competitive reverse transcription-polymerase chain reaction (RT-PCR) followed by southern blot. The template RNA was extracted from the splenocytes of rats infected with 20 metacercariae of P. westermani. The products of competitive RT-PCR were subjected to southern blot and enhanced chemiluminescence (ECL), and analyzed with a densitometer. In comparison with that of uninfected control rat splenocytes (value of 1), the levels of mRNA expression of IFN-γ had changed to 0.747 at 1 week post infection (PI), 0.00175 at 2 week PI, 0.0217 at 3 week PI, 0.194 at 4 week PI and then to 0.537 at 5 week PI. The level at 7 week PI had returned to 1.25, comparable with that of uninfected rats. These results show that, when infected with P. westermani, the levels of IFN-γ mRNA of SD rat splenocytes were remarkably reduced by more than 500 times at 2 week PI and restored to normal level at 7 week PI.

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Original Article

Changes in enzyme activity and expression of DHFR of Toxoplasma gondii by antifolates: Eun-Jung Baek, Ho-Woo Nam; Korean J Parasitol 1998;36(3):191-198.
Published online September 20, 1998; DOI: https://doi.org/10.3347/kjp.1998.36.3.191

Abstract
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The responses to antifolates of Toxoplasma gondii were investigated by measuring the dihydrofolate reductase (DHFR) activity, quantity of DHFR mRNA, and single-strand conformational polymorphism (SSCP) pattern. Pyrimethamine (PYM) and methotrexate (MTX) were tested as antifolates. When T. gondii was treated with PYM, the viability was decreased by the increasing concentration of PYM, DHFR activity tended to increase as the passage proceeded, and the quantity of mRNA expressed was also increased according to passages. The viability of T. gondii was decreased by the increasing concentration of MTX, but it was maintained over 40% up to 100 ?M MTX. DHFR activity was 77.4% in the 1st passage (1 ?M). 82.2% in the 4th passage (10 ?M), and 141.3% in the 7th passage (100 ?M). But no changes were detected in SSCP pattern of T. gondii exposed to PYM and MTX, both. These results suggested that the response of T. gondii to PYM was regulated by transcriptional level and that, in MTX, the viability of T. gondii was derived from increasing DHFR activity.

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