{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T23:47:29Z","timestamp":1790380049330,"version":"4.1.0"},"reference-count":66,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,3,13]],"date-time":"2020-03-13T00:00:00Z","timestamp":1584057600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,3,13]],"date-time":"2020-03-13T00:00:00Z","timestamp":1584057600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["098051"],"award-info":[{"award-number":["098051"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["098051"],"award-info":[{"award-number":["098051"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["098051"],"award-info":[{"award-number":["098051"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["098051"],"award-info":[{"award-number":["098051"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 AI045540"],"award-info":[{"award-number":["R01 AI045540"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006812","name":"Center for Integrated Healthcare, U.S. Department of Veterans Affairs","doi-asserted-by":"publisher","award":["BX001983"],"award-info":[{"award-number":["BX001983"]}],"id":[{"id":"10.13039\/100006812","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Visceral leishmaniasis is an infectious parasitic disease caused by the protozoan parasites<jats:italic>Leishmania donovani<\/jats:italic>and<jats:italic>Leishmania infantum<\/jats:italic>. The drugs currently used to treat visceral leishmaniasis suffer from toxicity and the emergence of parasite resistance, and so a better solution would be the development of an effective subunit vaccine; however, no approved vaccine currently exists. The comparative testing of a large number of vaccine candidates requires a quantitative and reproducible experimental murine infection model, but the parameters that influence infection pathology have not been systematically determined. To address this, we have established an infection model using a transgenic luciferase-expressing<jats:italic>L. donovani<\/jats:italic>parasite and longitudinally quantified the infections using<jats:italic>in vivo<\/jats:italic>bioluminescent imaging within individual mice. We examined the effects of varying the infection route, the site of adjuvant formulation administration, and standardised the parasite preparation and dose. We observed that the increase in parasite load within the liver during the first few weeks of infection was directly proportional to the parasite number in the initial inoculum. Finally, we show that immunity can be induced in pre-exposed animals that have resolved an initial infection. This murine infection model provides a platform for systematic subunit vaccine testing against visceral leishmaniasis.<\/jats:p>","DOI":"10.1038\/s41598-020-61662-3","type":"journal-article","created":{"date-parts":[[2020,3,13]],"date-time":"2020-03-13T11:03:20Z","timestamp":1584097400000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Establishment, optimisation and quantitation of a bioluminescent murine infection model of visceral leishmaniasis for systematic vaccine screening"],"prefix":"10.1038","volume":"10","author":[{"given":"Han Boon","family":"Ong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simon","family":"Clare","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam Jonathan","family":"Roberts","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary Edythe","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gavin James","family":"Wright","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,3,13]]},"reference":[{"key":"61662_CR1","unstructured":"WHO. Leishmaniasis. https:\/\/wwwwhoint\/news-room\/fact-sheets\/detail\/leishmaniasis. (2019)."},{"key":"61662_CR2","doi-asserted-by":"publisher","first-page":"e585","DOI":"10.1371\/journal.pntd.0000585","volume":"4","author":"C Gonz\u00e1lez","year":"2010","unstructured":"Gonz\u00e1lez, C. et al. Climate change and risk of leishmaniasis in north america: predictions from ecological niche models of vector and reservoir species. PLoS Negl. Trop. Dis. 4, e585, https:\/\/doi.org\/10.1371\/journal.pntd.0000585 (2010).","journal-title":"PLoS Negl. Trop. Dis."},{"key":"61662_CR3","doi-asserted-by":"publisher","first-page":"e0183583","DOI":"10.1371\/journal.pone.0183583","volume":"12","author":"BV Purse","year":"2017","unstructured":"Purse, B. V. et al. How will climate change pathways and mitigation options alter incidence of vector-borne diseases? A framework for leishmaniasis in South and Meso-America. PLoS One. 12, e0183583, https:\/\/doi.org\/10.1371\/journal.pone.0183583 (2017).","journal-title":"PLoS One."},{"key":"61662_CR4","doi-asserted-by":"publisher","first-page":"147","DOI":"10.2147\/CLEP.S44267","volume":"6","author":"PD Ready","year":"2014","unstructured":"Ready, P. D. Epidemiology of visceral leishmaniasis. Clin. Epidemiol. 6, 147\u201354, https:\/\/doi.org\/10.2147\/CLEP.S44267 (2014).","journal-title":"Clin. Epidemiol."},{"key":"61662_CR5","doi-asserted-by":"publisher","first-page":"925","DOI":"10.1016\/j.apjtm.2016.06.021","volume":"9","author":"A Oryan","year":"2016","unstructured":"Oryan, A. & Akbari, M. Worldwide risk factors in leishmaniasis. Asian Pac. J. Tropical Medicine. 9, 925\u201332, https:\/\/doi.org\/10.1016\/j.apjtm.2016.06.021 (2016).","journal-title":"Asian Pac. J. Tropical Medicine."},{"key":"61662_CR6","doi-asserted-by":"publisher","first-page":"20546","DOI":"10.2807\/1560-7917.ES2013.18.30.20546","volume":"18","author":"A Arce","year":"2013","unstructured":"Arce, A. et al. Re-emergence of leishmaniasis in Spain: community outbreak in Madrid, Spain, 2009 to 2012. Eurosurveillance. 18, 20546, https:\/\/doi.org\/10.2807\/1560-7917.ES2013.18.30.20546 (2013).","journal-title":"Eurosurveillance."},{"key":"61662_CR7","unstructured":"WHO. South Sudan intensifies measures against visceral leishmaniasis to improve health and social well-being of affected populations. https:\/\/wwwwhoint\/neglected_diseases\/news\/South-Sudan-intensifies-measures-against-VL\/en\/ (2018)."},{"key":"61662_CR8","doi-asserted-by":"crossref","unstructured":"Singh, O. P., Singh, B., Chakravarty, J. & Sundar, S. Current challenges in treatment options for visceral leishmaniasis in India: a public health perspective. Infect Dis Poverty. 5; 0.1186\/s40249-016-0112-2 (2016).","DOI":"10.1186\/s40249-016-0112-2"},{"key":"61662_CR9","doi-asserted-by":"publisher","first-page":"1530","DOI":"10.1093\/cid\/cit102","volume":"56","author":"S Rijal","year":"2013","unstructured":"Rijal, S. et al. Increasing failure of miltefosine in the treatment of kala-azar in nepal and the potential role of parasite drug resistance, reinfection, or noncompliance. Clin. Infect. Diseases. 56, 1530\u20138, https:\/\/doi.org\/10.1093\/cid\/cit102 (2013).","journal-title":"Clin. Infect. Diseases."},{"key":"61662_CR10","doi-asserted-by":"publisher","first-page":"9318","DOI":"10.1073\/pnas.1820175116","volume":"116","author":"S Wyllie","year":"2019","unstructured":"Wyllie, S. et al. Preclinical candidate for the treatment of visceral leishmaniasis that acts through proteasome inhibition. Proc. Natl Acad. Sciences. 116, 9318, https:\/\/doi.org\/10.1073\/pnas.1820175116 (2019).","journal-title":"Proc. Natl Acad. Sciences."},{"key":"61662_CR11","doi-asserted-by":"crossref","unstructured":"Wyllie, S. et al. Cyclin-dependent kinase 12 is a drug target for visceral leishmaniasis. Nature. 560, 192\u20137.10; 1038\/s41586-018-0356-z (2018).","DOI":"10.1038\/s41586-018-0356-z"},{"key":"61662_CR12","doi-asserted-by":"publisher","first-page":"3642","DOI":"10.1016\/j.vaccine.2005.02.015","volume":"23","author":"A Khamesipour","year":"2005","unstructured":"Khamesipour, A. et al. Leishmanization: Use of an old method for evaluation of candidate vaccines against leishmaniasis. Vaccine. 23, 3642\u20138, https:\/\/doi.org\/10.1016\/j.vaccine.2005.02.015 (2005).","journal-title":"Vaccine."},{"key":"61662_CR13","first-page":"1","volume":"6","author":"H Rezvan","year":"2015","unstructured":"Rezvan, H. & Moafi, M. An overview on Leishmania vaccines: A narrative review article. Vet. Res. Forum. 6, 1\u20137 (2015).","journal-title":"Vet. Res. Forum."},{"key":"61662_CR14","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jim.2015.03.017","volume":"422","author":"K Jain","year":"2015","unstructured":"Jain, K. & Jain, N. K. Vaccines for visceral leishmaniasis: A review. J. Immunological Methods. 422, 1\u201312, https:\/\/doi.org\/10.1016\/j.jim.2015.03.017 (2015).","journal-title":"J. Immunological Methods."},{"key":"61662_CR15","doi-asserted-by":"publisher","first-page":"123","DOI":"10.4103\/0019-5154.94280","volume":"57","author":"A Khamesipour","year":"2012","unstructured":"Khamesipour, A. et al. Treatment of cutaneous lesion of 20 years\u2019 duration caused by leishmanization. Indian. J. Dermatol. 57, 123\u20135, https:\/\/doi.org\/10.4103\/0019-5154.94280 (2012).","journal-title":"Indian. J. Dermatol."},{"key":"61662_CR16","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1093\/bmb\/ldl003","volume":"75-76","author":"MTM Roberts","year":"2006","unstructured":"Roberts, M. T. M. Current understandings on the immunology of leishmaniasis and recent developments in prevention and treatment. Br. Med. Bulletin. 75-76, 115\u201330, https:\/\/doi.org\/10.1093\/bmb\/ldl003 (2006).","journal-title":"Br. Med. Bulletin."},{"key":"61662_CR17","doi-asserted-by":"publisher","first-page":"1912","DOI":"10.4049\/jimmunol.166.3.1912","volume":"166","author":"PC Melby","year":"2001","unstructured":"Melby, P. C., Chandrasekar, B., Zhao, W. & Coe, J. E. The Hamster as a Model of Human Visceral Leishmaniasis: Progressive Disease and Impaired Generation of Nitric Oxide in the Face of a Prominent Th1-Like Cytokine Response. J. Immunology. 166, 1912, https:\/\/doi.org\/10.4049\/jimmunol.166.3.1912 (2001).","journal-title":"J. Immunology."},{"key":"61662_CR18","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1016\/j.vetpar.2012.10.002","volume":"192","author":"J Fern\u00e1ndez-Cotrina","year":"2013","unstructured":"Fern\u00e1ndez-Cotrina, J. et al. Experimental model for reproduction of canine visceral leishmaniosis by Leishmania infantum. Veterinary Parasitology. 192, 118\u201328, https:\/\/doi.org\/10.1016\/j.vetpar.2012.10.002 (2013).","journal-title":"Veterinary Parasitology."},{"key":"61662_CR19","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1590\/S0036-46652014000100001","volume":"56","author":"EN Lor\u00eda-Cervera","year":"2014","unstructured":"Lor\u00eda-Cervera, E. N. & Andrade-Narv\u00e1ez, F. J. Animal models for the study of leishmaniasis immunology. Rev. Inst. Med. Trop. Sao Paulo. 56, 1\u201311, https:\/\/doi.org\/10.1590\/S0036-46652014000100001 (2014).","journal-title":"Rev. Inst. Med. Trop. Sao Paulo."},{"key":"61662_CR20","first-page":"27","volume":"133","author":"S Gupta","year":"2011","unstructured":"Gupta, S. Nishi. Visceral leishmaniasis: experimental models for drug discovery. Indian. J. Med. Res. 133, 27\u201339 (2011).","journal-title":"Indian. J. Med. Res."},{"key":"61662_CR21","doi-asserted-by":"publisher","unstructured":"P\u00e9rez-Cabezas B, et al Understanding resistance versus susceptibility in visceral leishmaniasis using mouse models of Leishmania infantum infection. Frontiers in Cellular and Infection Microbiology. 9; https:\/\/doi.org\/10.3389\/fcimb.2019.00030 (2019).","DOI":"10.3389\/fcimb.2019.00030"},{"key":"61662_CR22","doi-asserted-by":"publisher","first-page":"3893","DOI":"10.1128\/iai.58.12.3893-3898.1990","volume":"58","author":"KE Squires","year":"1990","unstructured":"Squires, K. E. et al. Defect in the tissue cellular immune response: experimental visceral leishmaniasis in euthymic C57BL\/6 ep\/ep mice. Infect. Immunity. 58, 3893\u20138 (1990).","journal-title":"Infect. Immunity."},{"key":"61662_CR23","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1186\/s13071-016-1413-9","volume":"9","author":"C Loeuillet","year":"2016","unstructured":"Loeuillet, C., Ba\u00f1uls, A.-L. & Hide, M. Study of Leishmania pathogenesis in mice: experimental considerations. Parasit. Vectors. 9, 144, https:\/\/doi.org\/10.1186\/s13071-016-1413-9 (2016).","journal-title":"Parasit. Vectors."},{"key":"61662_CR24","doi-asserted-by":"publisher","first-page":"524","DOI":"10.1016\/j.pt.2004.08.009","volume":"20","author":"CR Engwerda","year":"2004","unstructured":"Engwerda, C. R., Ato, M. & Kaye, P. M. Macrophages, pathology and parasite persistence in experimental visceral leishmaniasis. Trends Parasitology. 20, 524\u201330, https:\/\/doi.org\/10.1016\/j.pt.2004.08.009 (2004).","journal-title":"Trends Parasitology."},{"key":"61662_CR25","doi-asserted-by":"publisher","first-page":"2958","DOI":"10.3389\/fimmu.2018.02958","volume":"9","author":"O Preham","year":"2018","unstructured":"Preham, O. et al. CD4(+) T Cells alter the stromal microenvironment and repress medullary erythropoiesis in murine visceral leishmaniasis. Front. Immunol. 9, 2958, https:\/\/doi.org\/10.3389\/fimmu.2018.02958 (2018).","journal-title":"Front. Immunol."},{"key":"61662_CR26","doi-asserted-by":"publisher","first-page":"e1006465","DOI":"10.1371\/journal.ppat.1006465","volume":"13","author":"AI Pinto","year":"2017","unstructured":"Pinto, A. I. et al. TNF signalling drives expansion of bone marrow CD4+ T cells responsible for HSC exhaustion in experimental visceral leishmaniasis. PLoS Pathog. 13, e1006465, https:\/\/doi.org\/10.1371\/journal.ppat.1006465 (2017).","journal-title":"PLoS Pathog."},{"key":"61662_CR27","doi-asserted-by":"publisher","first-page":"e0163604","DOI":"10.1371\/journal.pone.0163604","volume":"11","author":"JWJ Moore","year":"2016","unstructured":"Moore, J. W. J. et al. CD4+ Recent thymic emigrants are recruited into granulomas during Leishmania donovani infection but have limited capacity for cytokine production. PLoS one. 11, e0163604, https:\/\/doi.org\/10.1371\/journal.pone.0163604 (2016).","journal-title":"PLoS one."},{"key":"61662_CR28","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1016\/j.bbrc.2018.10.055","volume":"506","author":"P Srinontong","year":"2018","unstructured":"Srinontong, P., Wu, Z., Sato, K., Nagaoka, H. & Maekawa, Y. The circulating immunoglobulins negatively impact on the parasite clearance in the liver of Leishmania donovani-infected mice via dampening ROS activity. Biochemical Biophysical Res. Communications. 506, 20\u20136, https:\/\/doi.org\/10.1016\/j.bbrc.2018.10.055 (2018).","journal-title":"Biochemical Biophysical Res. Communications."},{"key":"61662_CR29","doi-asserted-by":"publisher","first-page":"2167","DOI":"10.1128\/aac.39.9.2167","volume":"39","author":"PA Buffet","year":"1995","unstructured":"Buffet, P. A., Sulahian, A., Garin, Y. J., Nassar, N. & Derouin, F. Culture microtitration: a sensitive method for quantifying Leishmania infantum in tissues of infected mice. Antimicrobial Agents Chemotherapy. 39, 2167\u20138, https:\/\/doi.org\/10.1128\/aac.39.9.2167 (1995).","journal-title":"Antimicrobial Agents Chemotherapy."},{"key":"61662_CR30","doi-asserted-by":"publisher","first-page":"e0185606","DOI":"10.1371\/journal.pone.0185606","volume":"12","author":"F Hossain","year":"2017","unstructured":"Hossain, F. et al. Real-time PCR in detection and quantitation of Leishmania donovani for the diagnosis of Visceral Leishmaniasis patients and the monitoring of their response to treatment. PLoS one. 12, e0185606, https:\/\/doi.org\/10.1371\/journal.pone.0185606 (2017).","journal-title":"PLoS one."},{"key":"61662_CR31","doi-asserted-by":"crossref","unstructured":"Santar\u00e9m, N., Tavares, J. & Cordeiro-da-Silva, A. In vitro infections of macrophage-like cell lines with Leishmania infantum for drug screening. In: Clos, J., editor. Leishmania: Methods and Protocols. New York, NY: Springer New York; p. 265\u201377. (2019).","DOI":"10.1007\/978-1-4939-9210-2_14"},{"key":"61662_CR32","doi-asserted-by":"publisher","first-page":"1059","DOI":"10.1038\/s41598-018-37670-9","volume":"9","author":"SL Berry","year":"2019","unstructured":"Berry, S. L., Walker, K., Hoskins, C., Telling, N. D. & Price, H. P. Nanoparticle-mediated magnetic hyperthermia is an effective method for killing the human-infective protozoan parasite Leishmania mexicana in vitro. Sci. Reports. 9, 1059, https:\/\/doi.org\/10.1038\/s41598-018-37670-9 (2019).","journal-title":"Sci. Reports."},{"key":"61662_CR33","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.ymeth.2017.05.008","volume":"127","author":"J Tavares","year":"2017","unstructured":"Tavares, J., Costa, D. M., Teixeira, A. R., Cordeiro-da-Silva, A. & Amino, R. In vivo imaging of pathogen homing to the host tissues. Methods. 127, 37\u201344, https:\/\/doi.org\/10.1016\/j.ymeth.2017.05.008 (2017).","journal-title":"Methods."},{"key":"61662_CR34","doi-asserted-by":"publisher","first-page":"383","DOI":"10.1111\/j.1462-5822.2004.00468.x","volume":"7","author":"T Lang","year":"2005","unstructured":"Lang, T., Goyard, S., Lebastard, M. & Milon, G. Bioluminescent Leishmania expressing luciferase for rapid and high throughput screening of drugs acting on amastigote-harbouring macrophages and for quantitative real-time monitoring of parasitism features in living mice. Cell. Microbiology. 7, 383\u201392, https:\/\/doi.org\/10.1111\/j.1462-5822.2004.00468.x (2005).","journal-title":"Cell. Microbiology."},{"key":"61662_CR35","doi-asserted-by":"publisher","first-page":"1980","DOI":"10.3791\/1980","volume":"41","author":"CJ Thalhofer","year":"2010","unstructured":"Thalhofer, C. J. et al. In vivo imaging of transgenic Leishmania parasites in a live host. J. Vis. Exp. 41, 1980, https:\/\/doi.org\/10.3791\/1980 (2010).","journal-title":"J. Vis. Exp."},{"key":"61662_CR36","doi-asserted-by":"publisher","first-page":"e0005924","DOI":"10.1371\/journal.pntd.0005924","volume":"11","author":"GD Melo","year":"2017","unstructured":"Melo, G. D. et al. New insights into experimental visceral leishmaniasis: Real-time in vivo imaging of Leishmania donovani virulence. PLoS Negl. Trop. Dis. 11, e0005924, https:\/\/doi.org\/10.1371\/journal.pntd.0005924 (2017).","journal-title":"PLoS Negl. Trop. Dis."},{"key":"61662_CR37","doi-asserted-by":"publisher","first-page":"933","DOI":"10.1016\/j.parint.2016.10.020","volume":"66","author":"E Rouault","year":"2017","unstructured":"Rouault, E. et al. Imaging visceral leishmaniasis in real time with golden hamster model: Monitoring the parasite burden and hamster transcripts to further characterize the immunological responses of the host. Parasitology International. 66, 933\u20139, https:\/\/doi.org\/10.1016\/j.parint.2016.10.020 (2017).","journal-title":"Parasitology International."},{"key":"61662_CR38","doi-asserted-by":"publisher","first-page":"e0007133","DOI":"10.1371\/journal.pntd.0007133","volume":"13","author":"R \u00c1lvarez-Velilla","year":"2019","unstructured":"\u00c1lvarez-Velilla, R. et al. A chronic bioluminescent model of experimental visceral leishmaniasis for accelerating drug discovery. PLoS Negl. Trop. Dis. 13, e0007133, https:\/\/doi.org\/10.1371\/journal.pntd.0007133 (2019).","journal-title":"PLoS Negl. Trop. Dis."},{"key":"61662_CR39","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1111\/j.1365-3024.2006.00817.x.","volume":"28","author":"J Carri\u00f3n","year":"2006","unstructured":"Carri\u00f3n, J. et al. Immunohistological features of visceral leishmaniasis in BALB\/c mice. Parasite Immunology. 28, 173\u201383, https:\/\/doi.org\/10.1111\/j.1365-3024.2006.00817.x. (2006).","journal-title":"Parasite Immunology."},{"key":"61662_CR40","doi-asserted-by":"publisher","first-page":"1413","DOI":"10.1007\/s00436-008-1150-x","volume":"103","author":"S Kaur","year":"2008","unstructured":"Kaur, S. et al. Effect of dose and route of inoculation on the generation of CD4+ Th1\/Th2 type of immune response in murine visceral leishmaniasis. Parasitology Research. 103, 1413\u20139, https:\/\/doi.org\/10.1007\/s00436-008-1150-x (2008).","journal-title":"Parasitology Research."},{"key":"61662_CR41","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1016\/j.actatropica.2003.09.010","volume":"90","author":"N Rol\u00e3o","year":"2004","unstructured":"Rol\u00e3o, N., Melo, C. & Campino, L. Influence of the inoculation route in BALB\/c mice infected by Leishmania infantum. Acta Tropica. 90, 123\u20136, https:\/\/doi.org\/10.1016\/j.actatropica.2003.09.010 (2004).","journal-title":"Acta Tropica."},{"key":"61662_CR42","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/S0165-2478(03)00021-X","volume":"86","author":"P Mukherjee","year":"2003","unstructured":"Mukherjee, P., Ghosh, A. K. & Ghose, A. C. Infection pattern and immune response in the spleen and liver of BALB\/c mice intracardially infected with Leishmania donovani amastigotes. Immunology Letters. 86, 131\u20138, https:\/\/doi.org\/10.1016\/S0165-2478(03)00021-X (2003).","journal-title":"Immunology Letters."},{"key":"61662_CR43","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1006\/expr.2001.4656","volume":"99","author":"GF Sp\u00e4th","year":"2001","unstructured":"Sp\u00e4th, G. F. & Beverley, S. M. A Lipophosphoglycan-Independent Method for Isolation of Infective Leishmania Metacyclic Promastigotes by Density Gradient Centrifugation. Exp. Parasitology. 99, 97\u2013103, https:\/\/doi.org\/10.1006\/expr.2001.4656 (2001).","journal-title":"Exp. Parasitology."},{"key":"61662_CR44","doi-asserted-by":"publisher","first-page":"3976","DOI":"10.1074\/mcp.O113.028357","volume":"12","author":"C Crosnier","year":"2013","unstructured":"Crosnier, C. et al. A library of functional recombinant cell-surface and secreted P. falciparum merozoite proteins. Mol. Cell. Proteomics. 12, 3976\u201386, https:\/\/doi.org\/10.1074\/mcp.O113.028357 (2013).","journal-title":"Mol. Cell. Proteomics."},{"key":"61662_CR45","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1016\/0014-4894(89)90122-7","volume":"68","author":"KC Carter","year":"1989","unstructured":"Carter, K. C., Alexander, J., Baillie, A. J. & Dolan, T. F. Visceral leishmaniasis: Resistance to reinfection in the liver following chemotherapy in the BALB\/c mouse. Exp. Parasitology. 68, 375\u201381, https:\/\/doi.org\/10.1016\/0014-4894(89)90122-7 (1989).","journal-title":"Exp. Parasitology."},{"key":"61662_CR46","doi-asserted-by":"publisher","first-page":"5486","DOI":"10.4049\/jimmunol.176.9.5486","volume":"176","author":"M Ato","year":"2006","unstructured":"Ato, M. et al. Loss of dendritic cell migration and impaired resistance to Leishmania donovani infection in mice deficient in CCL19 and CCL21. J. immunology 176, 5486\u201393 (2006).","journal-title":"J. immunology"},{"key":"61662_CR47","doi-asserted-by":"publisher","first-page":"1927","DOI":"10.1084\/jem.167.6.1927","volume":"167","author":"MJ McElrath","year":"1988","unstructured":"McElrath, M. J., Murray, H. W. & Cohn, Z. A. The dynamics of granuloma formation in experimental visceral leishmaniasis. J. Exp. Medicine. 167, 1927\u201337, https:\/\/doi.org\/10.1084\/jem.167.6.1927 (1988).","journal-title":"J. Exp. Medicine."},{"key":"61662_CR48","doi-asserted-by":"crossref","first-page":"2231","DOI":"10.4049\/jimmunol.156.6.2231","volume":"156","author":"ME Wilson","year":"1996","unstructured":"Wilson, M. E. et al. Local suppression of IFN-gamma in hepatic granulomas correlates with tissue-specific replication of Leishmania chagasi. J. Immunology. 156, 2231\u20139 (1996).","journal-title":"J. Immunology."},{"key":"61662_CR49","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1128\/CMR.18.2.293-305.2005","volume":"18","author":"M Olivier","year":"2005","unstructured":"Olivier, M., Gregory, D. J. & Forget, G. Subversion mechanisms by which Leishmania parasites can escape the host immune response: a signaling point of view. Clin. Microbiol. Rev. 18, 293\u2013305, https:\/\/doi.org\/10.1128\/CMR.18.2.293-305.2005 (2005).","journal-title":"Clin. Microbiol. Rev."},{"key":"61662_CR50","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1016\/0014-4894(89)90176-8","volume":"69","author":"DL Sacks","year":"1989","unstructured":"Sacks, D. L. Metacyclogenesis in Leishmania promastigotes. Exp. Parasitology. 69, 100\u20133, https:\/\/doi.org\/10.1016\/0014-4894(89)90176-8 (1989).","journal-title":"Exp. Parasitology."},{"key":"61662_CR51","doi-asserted-by":"publisher","first-page":"2802","DOI":"10.1128\/iai.55.11.2802-2806.1987","volume":"55","author":"R da Silva","year":"1987","unstructured":"da Silva, R. & Sacks, D. L. Metacyclogenesis is a major determinant of Leishmania promastigote virulence and attenuation. Infect. immunity. 55, 2802\u20136 (1987).","journal-title":"Infect. immunity."},{"key":"61662_CR52","doi-asserted-by":"publisher","first-page":"1041","DOI":"10.1038\/ni1102-1041","volume":"3","author":"D Sacks","year":"2002","unstructured":"Sacks, D. & Sher, A. Evasion of innate immunity by parasitic protozoa. Nat. Immunology. 3, 1041\u20137, https:\/\/doi.org\/10.1038\/ni1102-1041 (2002).","journal-title":"Nat. Immunology."},{"key":"61662_CR53","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1046\/j.1365-2958.1998.00689.x","volume":"27","author":"PB Joshi","year":"1998","unstructured":"Joshi, P. B., Sacks, D. L., Modi, G. & McMaster, W. R. Targeted gene deletion of Leishmania major genes encoding developmental stage-specific leishmanolysin (GP63). Mol. Microbiology. 27, 519\u201330, https:\/\/doi.org\/10.1046\/j.1365-2958.1998.00689.x (1998).","journal-title":"Mol. Microbiology."},{"key":"61662_CR54","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1046\/j.1462-5822.2001.00135.x","volume":"3","author":"PG McKean","year":"2001","unstructured":"McKean, P. G., Denny, P. W., Knuepfer, E., Keen, J. K. & Smith, D. F. Phenotypic changes associated with deletion and overexpression of a stage-regulated gene family in Leishmania. Cell. Microbiology. 3, 511\u201323, https:\/\/doi.org\/10.1046\/j.1462-5822.2001.00135.x (2001).","journal-title":"Cell. Microbiology."},{"key":"61662_CR55","doi-asserted-by":"publisher","first-page":"e1833","DOI":"10.1371\/journal.pntd.0001833","volume":"6","author":"TD Serafim","year":"2012","unstructured":"Serafim, T. D. et al. Leishmania metacyclogenesis is promoted in the absence of purines. PLoS Negl. Trop. Dis. 6, e1833, https:\/\/doi.org\/10.1371\/journal.pntd.0001833 (2012).","journal-title":"PLoS Negl. Trop. Dis."},{"key":"61662_CR56","doi-asserted-by":"publisher","first-page":"285","DOI":"10.1126\/science.1057740","volume":"292","author":"ML Cunningham","year":"2001","unstructured":"Cunningham, M. L., Titus, R. G., Turco, S. J. & Beverley, S. M. Regulation of differentiation to the infective stage of the protozoan parasite Leishmania major by tetrahydrobiopterin. Science. 292, 285, https:\/\/doi.org\/10.1126\/science.1057740 (2001).","journal-title":"Science."},{"key":"61662_CR57","doi-asserted-by":"publisher","first-page":"13837","DOI":"10.1073\/pnas.0600843103","volume":"103","author":"G van Zandbergen","year":"2006","unstructured":"van Zandbergen, G. et al. Leishmania disease development depends on the presence of apoptotic promastigotes in the virulent inoculum. Proc. Natl Acad. Sciences. 103, 13837, https:\/\/doi.org\/10.1073\/pnas.0600843103 (2006).","journal-title":"Proc. Natl Acad. Sciences."},{"key":"61662_CR58","doi-asserted-by":"publisher","first-page":"e65960","DOI":"10.1371\/journal.pone.0065960","volume":"8","author":"SH Sheehy","year":"2013","unstructured":"Sheehy, S. H. et al. Optimising controlled human malaria infection studies using cryopreserved P. falciparum parasites administered by needle and syringe. PLoS one. 8, e65960, https:\/\/doi.org\/10.1371\/journal.pone.0065960 (2013).","journal-title":"PLoS one."},{"key":"61662_CR59","doi-asserted-by":"publisher","first-page":"502","DOI":"10.1038\/nature01152","volume":"420","author":"Y Belkaid","year":"2002","unstructured":"Belkaid, Y., Piccirillo, C. A., Mendez, S., Shevach, E. M. & Sacks, D. L. CD4+CD25+ regulatory T cells control Leishmania major persistence and immunity. Nature. 420, 502\u20137, https:\/\/doi.org\/10.1038\/nature01152 (2002).","journal-title":"Nature."},{"key":"61662_CR60","doi-asserted-by":"publisher","first-page":"114","DOI":"10.3389\/fimmu.2013.00114","volume":"4","author":"S Awate","year":"2013","unstructured":"Awate, S., Babiuk, L. A. & Mutwiri, G. Mechanisms of action of adjuvants. Front. Immunol. 4, 114, https:\/\/doi.org\/10.3389\/fimmu.2013.00114 (2013).","journal-title":"Front. Immunol."},{"key":"61662_CR61","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1016\/j.femsim.2004.05.008","volume":"42","author":"P Tewary","year":"2004","unstructured":"Tewary, P., Mehta, J., Sukumaran, B. & Madhubala, R. Vaccination with Leishmania soluble antigen and immunostimulatory oligodeoxynucleotides induces specific immunity and protection against Leishmania donovani infection. FEMS Immunology Med. Microbiology. 42, 241\u20138, https:\/\/doi.org\/10.1016\/j.femsim.2004.05.008 (2004).","journal-title":"FEMS Immunology Med. Microbiology."},{"key":"61662_CR62","doi-asserted-by":"publisher","first-page":"14006","DOI":"10.1074\/jbc.M610869200","volume":"282","author":"AA Capul","year":"2007","unstructured":"Capul, A. A., Barron, T., Dobson, D. E., Turco, S. J. & Beverley, S. M. Two functionally divergent UDP-Gal nucleotide sugar transporters participate in phosphoglycan synthesis in Leishmania major. J. Biol. Chem. 282, 14006\u201317, https:\/\/doi.org\/10.1074\/jbc.M610869200 (2007).","journal-title":"J. Biol. Chem."},{"key":"61662_CR63","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1016\/0166-6851(91)90210-W","volume":"46","author":"CM Coburn","year":"1991","unstructured":"Coburn, C. M., Otteman, K. M., McNeely, T., Turco, S. J. & Beverley, S. M. Stable DNA transfection of a wide range of trypanosomatids. Mol. Biochemical Parasitology. 46, 169\u201379, https:\/\/doi.org\/10.1016\/0166-6851(91)90210-W (1991).","journal-title":"Mol. Biochemical Parasitology."},{"key":"61662_CR64","doi-asserted-by":"crossref","first-page":"5505","DOI":"10.4049\/jimmunol.163.10.5505","volume":"163","author":"A St-Denis","year":"1999","unstructured":"St-Denis, A., Caouras, V., Gervais, F. & Descoteaux, A. Role of Protein Kinase C-\u03b1 in the Control of Infection by Intracellular Pathogens in Macrophages. J. Immunology. 163, 5505\u201311 (1999).","journal-title":"J. Immunology."},{"key":"61662_CR65","doi-asserted-by":"publisher","first-page":"364","DOI":"10.1016\/j.actatropica.2006.01.004","volume":"97","author":"S Wyllie","year":"2006","unstructured":"Wyllie, S. & Fairlamb, A. H. Refinement of techniques for the propagation of Leishmania donovani in hamsters. Acta Tropica. 97, 364\u20139, https:\/\/doi.org\/10.1016\/j.actatropica.2006.01.004 (2006).","journal-title":"Acta Tropica."},{"key":"61662_CR66","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1016\/j.ijpara.2018.10.003","volume":"49","author":"CM Elton","year":"2019","unstructured":"Elton, C. M., Rodriguez, M., Ben Mamoun, C., Lobo, C. A. & Wright, G. J. A library of recombinant Babesia microti cell surface and secreted proteins for diagnostics discovery and reverse vaccinology. Int. J. Parasitol. 49, 115\u201325, https:\/\/doi.org\/10.1016\/j.ijpara.2018.10.003 (2019).","journal-title":"Int. J. Parasitol."}],"updated-by":[{"DOI":"10.1038\/s41598-021-87190-2","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2021,4,7]],"date-time":"2021-04-07T00:00:00Z","timestamp":1617753600000}}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61662-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61662-3","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61662-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,28]],"date-time":"2023-09-28T16:13:32Z","timestamp":1695917612000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61662-3"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,13]]},"references-count":66,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["61662"],"URL":"https:\/\/doi.org\/10.1038\/s41598-020-61662-3","relation":{"correction":[{"id-type":"doi","id":"10.1038\/s41598-021-87190-2","asserted-by":"object"}]},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,13]]},"assertion":[{"value":"14 January 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"27 February 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"13 March 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 April 2021","order":4,"name":"change_date","label":"Change Date","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Correction","order":5,"name":"change_type","label":"Change Type","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"A Correction to this paper has been published:","order":6,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"https:\/\/doi.org\/10.1038\/s41598-021-87190-2","URL":"https:\/\/doi.org\/10.1038\/s41598-021-87190-2","order":7,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"4689"}}