{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:57:20Z","timestamp":1789693040300,"version":"4.0.1"},"reference-count":40,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,1,14]],"date-time":"2021-01-14T00:00:00Z","timestamp":1610582400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81802037"],"award-info":[{"award-number":["81802037"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81871684"],"award-info":[{"award-number":["81871684"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004731","name":"Natural Science Foundation of Zhejiang Province","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004731","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p><jats:italic>Toxoplasma gondii<\/jats:italic>is an obligate intracellular protozoan parasite.<jats:italic>T. gondii<\/jats:italic>primarily infection in pregnant women may result in fetal abortion, and infection in immunosuppressed population may result in toxoplasmosis. Carbamoyl phosphate synthetase II (CPSII) is a key enzyme in the<jats:italic>de novo<\/jats:italic>pyrimidine-biosynthesis pathway, and has a crucial role in parasite replication. We generated a mutant with complete deletion of CPSII via clustered regularly interspaced short palindromic repeats (CRISPR)\/cas9 in type-1 RH strain of<jats:italic>T. gondii<\/jats:italic>. We tested the intracellular proliferation of this mutant and found that it showed significantly reduced replication<jats:italic>in vitro<\/jats:italic>, though CPSII deletion did not completely stop the parasite growth. The immune responses induced by the infection of RH\u0394CPSII tachyzoites in mice were evaluated. During infection in mice, the RH\u0394CPSII mutant displayed notable defects in replication and virulence, and significantly enhanced the survival of mice compared with survival of RH-infected mice. We tracked parasite propagation from ascitic fluid in mice infected with the RH\u0394CPSII mutant, and few tachyzoites were observed at early infection. We also observed that the RH\u0394CPSII mutant induced greater accumulation of neutrophils. The mutant induced a higher level of T-helper type-1 cytokines [interferon (IFN)-\u03b3, interleukin (IL)-12]. The mRNA levels of signal transducer and activator of transcription cellular transcription factor 1 and IFN regulatory factor 8 were significantly higher in the RH\u0394CPSII mutant-infected group. Together, these data suggest that CPSII is crucial for parasite growth, and that strains lack the<jats:italic>de novo<\/jats:italic>pyrimidine biosynthesis pathway and salvage pathway may become a promising live attenuated vaccine to prevent infection with<jats:italic>T. gondii<\/jats:italic>.<\/jats:p>","DOI":"10.3389\/fmicb.2020.616688","type":"journal-article","created":{"date-parts":[[2021,1,14]],"date-time":"2021-01-14T18:04:34Z","timestamp":1610647474000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["A Carbamoyl Phosphate Synthetase II (CPSII) Deletion Mutant of Toxoplasma gondii Induces Partial Protective Immunity in Mice"],"prefix":"10.3389","volume":"11","author":[{"given":"Xunhui","family":"Zhuo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaige","family":"Du","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haojie","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Di","family":"Lou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaohong","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,1,14]]},"reference":[{"key":"B1","doi-asserted-by":"crossref","first-page":"6361","DOI":"10.1073\/pnas.1816161116","article-title":"Divergent kinase regulates membrane ultrastructure of the Toxoplasma parasitophorous vacuole.","volume":"116","author":"Beraki","year":"2019","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"B2","doi-asserted-by":"crossref","DOI":"10.12688\/f1000research.16021.1","article-title":"Metabolic interactions between Toxoplasma gondii and its host.","volume":"7","author":"Blume","year":"2018","journal-title":"F1000Research"},{"key":"B3","doi-asserted-by":"publisher","first-page":"5216","DOI":"10.1128\/iai.70.9.5216-5224.2002","article-title":"Mechanism of entry determines the ability of Toxoplasma gondii to inhibit macrophage proinflammatory cytokine production.","volume":"70","author":"Butcher","year":"2002","journal-title":"Infect. Immun."},{"key":"B4","doi-asserted-by":"publisher","first-page":"637","DOI":"10.1089\/fpd.2017.2305","article-title":"Toxoplasma gondii, a Foodborne Pathogen in the Swine Production Chain from a European Perspective.","volume":"14","author":"De Berardinis","year":"2017","journal-title":"Foodborne Pathogens Dis."},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2019.111711","article-title":"Recent progress on anti-Toxoplasma drugs discovery: Design, synthesis and screening.","volume":"183","author":"Deng","year":"2019","journal-title":"Eur. J. Med. Chem."},{"key":"B6","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1017\/s0031182019001586","article-title":"Genotyping of viable Toxoplasma gondii from the first national survey of feral swine revealed evidence for sylvatic transmission cycle, and presence of highly virulent parasite genotypes.","volume":"147","author":"Dubey","year":"2020","journal-title":"Parasitology"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004047","article-title":"Parasite fate and involvement of infected cells in the induction of CD4 + and CD8 + T cell responses to Toxoplasma gondii.","volume":"10","author":"Dupont","year":"2014","journal-title":"PLoS pathogens"},{"key":"B8","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1016\/j.pt.2010.01.009","article-title":"Toxoplasma gondii: epidemiology, feline clinical aspects, and prevention.","volume":"26","author":"Elmore","year":"2010","journal-title":"Trends Parasitol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"4","DOI":"10.7774\/cevr.2019.8.1.4","article-title":"Rhoptry antigens as Toxoplasma gondii vaccine target.","volume":"8","author":"Foroutan","year":"2019","journal-title":"Clin. Exp. Vacc. Res."},{"key":"B10","doi-asserted-by":"publisher","first-page":"926","DOI":"10.1038\/415926a","article-title":"De novo pyrimidine biosynthesis is required for virulence of Toxoplasma gondii.","volume":"415","author":"Fox","year":"2002","journal-title":"Nature"},{"key":"B11","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1016\/s0020-7519(02)00214-x","article-title":"Organisation and sequence determination of glutamine-dependent carbamoyl phosphate synthetase II in Toxoplasma gondii.","volume":"33","author":"Fox","year":"2003","journal-title":"Int. J. Parasitol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"3744","DOI":"10.1128\/iai.00287-10","article-title":"Avirulent uracil auxotrophs based on disruption of orotidine-5\u2032-monophosphate decarboxylase elicit protective immunity to Toxoplasma gondii.","volume":"78","author":"Fox","year":"2010","journal-title":"Infect. Immun."},{"key":"B13","doi-asserted-by":"publisher","first-page":"2148","DOI":"10.1128\/iai.02756-14","article-title":"Non-replicating, cyst-defective type II Toxoplasma gondii vaccine strains stimulate protective immunity against acute and chronic infection.","volume":"83","author":"Fox","year":"2015","journal-title":"Infect. Immun."},{"key":"B14","doi-asserted-by":"publisher","first-page":"533","DOI":"10.1016\/j.ijpara.2008.09.011","article-title":"Genetic identification of essential indels and domains in carbamoyl phosphate synthetase II of Toxoplasma gondii.","volume":"39","author":"Fox","year":"2009","journal-title":"Int. J. Parasitol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"1155","DOI":"10.1016\/j.cmi.2020.04.014","article-title":"Toxoplasma gondii-specific IgG avidity testing in pregnant women.","author":"Garnaud","year":"2020","journal-title":"Clin. Microbiol. Infect."},{"key":"B16","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1016\/j.vetpar.2018.02.043","article-title":"Assessment of the antigenic and neuroprotective activity of the subunit anti-Toxoplasma vaccine in T. gondii experimentally infected mice.","volume":"254","author":"Gatkowska","year":"2018","journal-title":"Vet. Parasitol."},{"key":"B17","doi-asserted-by":"crossref","first-page":"2533","DOI":"10.4049\/jimmunol.153.6.2533","article-title":"Parasite-induced IL-12 stimulates early IFN-gamma synthesis and resistance during acute infection with Toxoplasma gondii.","volume":"153","author":"Gazzinelli","year":"1994","journal-title":"J. Immunol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"1069","DOI":"10.4049\/jimmunol.182.2.1069","article-title":"Cell-mediated immunity to Toxoplasma gondii develops primarily by local Th1 host immune responses in the absence of parasite replication.","volume":"182","author":"Gigley","year":"","journal-title":"J. Immunol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"5380","DOI":"10.1128\/iai.00649-09","article-title":"Long-term immunity to lethal acute or chronic type II Toxoplasma gondii infection is effectively induced in genetically susceptible C57BL\/6 mice by immunization with an attenuated type I vaccine strain.","volume":"77","author":"Gigley","year":"","journal-title":"Infect. Immun."},{"key":"B20","doi-asserted-by":"publisher","first-page":"174","DOI":"10.5455\/javar.2019.f329","article-title":"Epitope-based vaccine as a universal vaccination strategy against Toxoplasma gondii infection: A mini-review.","volume":"6","author":"Hajissa","year":"2019","journal-title":"J. Adv. Vet. Anim. Res."},{"key":"B21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/microbiolspec.PFS-0011-2015","article-title":"Toxoplasma gondii as a Parasite in Food: Analysis and Control.","volume":"4","author":"Hill","year":"2016","journal-title":"Microbiol. Spect."},{"key":"B22","doi-asserted-by":"publisher","first-page":"2974","DOI":"10.1128\/iai.00187-16","article-title":"Pyrimidine Pathway-Dependent and -Independent Functions of the Toxoplasma gondii Mitochondrial Dihydroorotate Dehydrogenase.","volume":"84","author":"Hortua Triana","year":"2016","journal-title":"Infect. Immun."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00196","article-title":"Innate Lymphoid Cells in Protection, Pathology, and Adaptive Immunity During Apicomplexan Infection.","volume":"10","author":"Ivanova","year":"2019","journal-title":"Front. Immunol."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00325","article-title":"Regulating IRFs in IFN Driven Disease.","volume":"10","author":"Jefferies","year":"2019","journal-title":"Front. Immunol."},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2017.00029","article-title":"Canonical and Non-Canonical Aspects of JAK-STAT Signaling: Lessons from Interferons for Cytokine Responses.","volume":"8","author":"Majoros","year":"2017","journal-title":"Front. Immunol."},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.02587","article-title":"Drug Resistance in Toxoplasma gondii.","volume":"9","author":"Montazeri","year":"2018","journal-title":"Front. Microbiol."},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2020.105544","article-title":"Risk factors associated with seropositivity for Toxoplasma gondii in population-based studies among immunocompromised patients (pregnant women, HIV patients and children) in West African countries, Cameroon and Gabon: a meta-analysis.","volume":"209","author":"Odeniran","year":"2020","journal-title":"Acta Tropica"},{"key":"B28","doi-asserted-by":"publisher","first-page":"388","DOI":"10.1016\/j.pt.2011.03.009","article-title":"Innate responses to Toxoplasma gondii in mice and humans.","volume":"27","author":"Pifer","year":"2011","journal-title":"Trends Parasitol."},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.02223","article-title":"QuilA-Adjuvanted T. gondii Lysate Antigens Trigger Robust Antibody and IFNgamma(+) T Cell Responses in Pigs Leading to Reduction in Parasite DNA in Tissues Upon Challenge Infection.","volume":"10","author":"Rahman","year":"2019","journal-title":"Front. Immunol."},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2020.00161","article-title":"Early Kinetics of Intestinal Infection and Immune Responses to Two Toxoplasma gondii Strains in Pigs.","volume":"10","author":"Rahman","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1007\/978-1-4939-6472-7_6","article-title":"Development of CRISPR\/Cas9 for Efficient Genome Editing in Toxoplasma gondii.","volume":"1498","author":"Shen","year":"2017","journal-title":"Methods Mole. Biol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"10711","DOI":"10.1073\/pnas.1307868110","article-title":"TLR-independent neutrophil-derived IFN-gamma is important for host resistance to intracellular pathogens.","volume":"110","author":"Sturge","year":"2013","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0216615","article-title":"A social cost-benefit analysis of two One Health interventions to prevent toxoplasmosis.","volume":"14","author":"Suijkerbuijk","year":"2019","journal-title":"PLoS One"},{"key":"B34","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1016\/0264-410x(90)90071-s","article-title":"Development of Toxoplasma gondii vaccine: A global challenge.","volume":"9","author":"Verma","year":"2013","journal-title":"Hum. Vac. Immunother."},{"key":"B35","doi-asserted-by":"publisher","first-page":"1562","DOI":"10.4049\/jimmunol.1900410","article-title":"Toxoplasma gondii tkl1 Deletion Mutant Is a Promising Vaccine against Acute, Chronic, and Congenital Toxoplasmosis in Mice.","volume":"204","author":"Wang","year":"2020","journal-title":"J. Immunol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1016\/j.pt.2019.01.005","article-title":"Advances in the Development of Anti-Toxoplasma gondii Vaccines: Challenges, Opportunities, and Perspectives.","volume":"35","author":"Wang","year":"2019","journal-title":"Trends Parasitol."},{"key":"B37","doi-asserted-by":"crossref","first-page":"e177","DOI":"10.1016\/S2352-3018(17)30005-X","article-title":"Prevalence and burden of Toxoplasma gondii infection in HIV-infected people: a systematic review and meta-analysis.","volume":"4","author":"Wang","year":"2017","journal-title":"Lancet. HIV"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2019.105148","article-title":"Vaccination with recombinant Toxoplasma gondii CDPK3 induces protective immunity against experimental toxoplasmosis.","volume":"199","author":"Wu","year":"2019","journal-title":"Acta Tropica"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.01814","article-title":"Mutant of Toxoplasma Stimulates Protective Immunity Against Acute and Chronic Toxoplasmosis.","volume":"9","author":"Xia","year":"2018","journal-title":"Front. Immunol."},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00418","article-title":"GRA24-Based DNA Vaccine Prolongs Survival in Mice Challenged With a Virulent Toxoplasma gondii Strain.","volume":"10","author":"Zheng","year":"2019","journal-title":"Front. Immunol."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2020.616688\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,17]],"date-time":"2023-10-17T18:00:21Z","timestamp":1697565621000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2020.616688\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,14]]},"references-count":40,"alternative-id":["10.3389\/fmicb.2020.616688"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2020.616688","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,14]]},"article-number":"616688"}}