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Elucidating in vitro and in vivo phenotypic behaviour of L. infantum/L. major natural hybrids

  • S. Cortes (a1), A. Albuquerque-Wendt (a1), C. Maia (a1), M. Carvalho (a1) (a2), I. A. Lima (a3), L. A. R. de Freitas (a3), W. L. C. dos-Santos (a3) and L. Campino (a1)...


The clinical manifestation and course of Leishmania infections depend on factors such as species, virulence and host-immunity. Although trypanosomatids are considered to have clonal propagation, genetic hybridization has produced successful natural hybrid lineages. Hybrids displaying strong selective advantages may have an impact on pathogenesis and the eco-epidemiology of leishmaniasis. Thus, characterization of phenotypic properties of Leishmania hybrids could bring significant insight into the biology, infectivity, pathogenicity and transmission dynamics of these atypical strains. The present study focuses on phenotypic features and survival capacity of Leishmania infantum/Leishmania major hybrid isolates as compared with representative putative parental species, L. infantum and L. major. In vitro assays (growth kinetics, susceptibility to different conditions) and in vivo infection (parasite detection and histopathological alterations) showed that hybrids present higher growth capacity and decreased susceptibility to reactive oxygen species. Furthermore, evaluation of infected spleen tissue suggests that hybrids induce a stronger immune reaction than their putative parents, leading to the development of white pulp hyperplasia in B-lymphocyte compartments. Overall, these hybrids have shown high plasticity in terms of their general behaviour within the different phenotypic parameters, suggesting that they might have acquired genetic features conferring different mechanisms to evade host cells.


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Author for correspondence: S. Cortes, E-mail:


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These authors contributed equally to this work.

Present address: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, OX1 3RE, Oxford, UK



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Akopyants, NS, Kimblin, N, Secundino, N, Patrick, R, Peters, N, Lawyer, P, Dobson, DE, Beverley, SM and Sacks, DL (2009) Demonstration of genetic exchange during cyclical development of Leishmania in the sand fly vector. Science 324, 265.10.1126/science.1169464
Almeida, MC, Cuba, CA, de Sa, CM, Pharoah, MM, Howard, KM and Miles, MA (1993) Metacyclogenesis of Leishmania (viannia) braziliensis in vitro: evidence that lentil lectin is a marker of complement resistance and enhanced infectivity. Transactions of the Royal Society of Tropical Medicine and Hygiene 87, 325329.10.1016/0035-9203(93)90150-O
Bañuls, AL, Jonquieres, R, Guerrini, F, Le Pont, F, Barrera, C, Espinel, I, Guderian, R, Echeverria, R and Tibayrenc, M (1999) Genetic analysis of Leishmania parasites in Ecuador: are Leishmania (viannia) panamensis and Leishmania (v.) Guyanensis distinct taxa? American Society of Tropical Medicine and Hygiene 61, 838845.
Belli, AA, Miles, MA and Kelly, JM (1994) A putative Leishmania panamensis/leishmania braziliensis hybrid is a causative agent of human cutaneous leishmaniasis in Nicaragua. Parasitology 109(Pt4), 435442.
Calvo-Álvarez, E, Álvarez-Velilla, R, Jiménez, M, Molina, R, Pérez-Pertejo, Y, Balaña-Fouce, R and Reguera, RM (2014) First evidence of intraclonal genetic exchange in trypanosomatids using two Leishmania infantum fluorescent transgenic clones. PLoS Neglected Tropical Diseases 8, e3075.10.1371/journal.pntd.0003075
Chargui, N, Amro, A, Haouas, N, Schönian, G, Babba, H, Schmidt, S, Ravel, C, Lefebvre, M, Bastien, P, Chaker, E, Aoun, K, Zribi, M and Kuhls, K (2009) Population structure of Tunisian Leishmania infantum and evidence for the existence of hybrids and gene flow between genetically different populations. International Journal for Parasitology 39, 801811.10.1016/j.ijpara.2008.11.016
Chemkhi, J, Souguir, H, Ali, IB, Driss, M, Guizani, I and Guerbouj, S (2015) Natural infection of Algerian hedgehog, Atelerix algirus (Lereboullet 1842) with Leishmania parasites in Tunisia. Acta Tropica 150, 4251.
Ciháková, J and Volf, P (1997) Development of different Leishmania major strains in the vector sandflies Phlebotomus papatasi and P. duboscqi. Annals of tropical medicine and parasitology 91, 267279.
Cortes, S, Esteves, C, Maurício, I, Maia, C, Cristovão, JM, Miles, M and Campino, L (2012) In vitro and in vivo behaviour of sympatric Leishmania (v.) braziliensis, L. (V.) peruviana and their hybrids. Parasitology 139, 191199.
De Lima, VM, Fattori, KR, de Souza, F, Eugenio, FR, dos Santos, PS, Rozza, DB and Machado, GF (2012) Apoptosis in T lymphocytes from spleen tissue and peripheral blood of L. (L.) chagasi naturally infected dogs. Veterinary Parasitology 184, 147153.
Delgado, O, Cupolillo, E, Bonfante-Garrido, R, Silva, S, Belfort, E, Grimaldi Júnior, G and Momen, H (1997) Cutaneous Leishmaniasis in Venezuela caused by infection with a new hybrid between Leishmania (viannia) braziliensis and l. (V.) guyanensis. Memórias Instituto Oswaldo Cruz 92, 581582.
Dujardin, JC, Bañuls, AL, Llanos-Cuentas, A, Alvarez, E, De Doncker, S, Jacquet, D, Le Ray, D, Arevalo, J and Tibayrenc, M (1995) Putative Leishmania hybrids in the Eastern Andean valley of Huanuco, Peru. Acta Tropica 59, 293307.
El Tai, NO, Osman, OF, el Fari, M, Presber, W and Schönian, G (2000) Genetic heterogeneity of ribosomal internal transcribed spacer in clinical samples of Leishmania donovani spotted on filter paper as revealed by single-strand conformation polymorphisms and sequencing. Transactions of the Royal Society of Tropical Medicine and Hygiene 94, 575579.
Garin, YJ, Sulahian, A, Pratlong, F, Meneceur, P, Gangneux, JP, Prina, E, Dedet, JP and Derouin, F (2001) Virulence of Leishmania infantum is expressed as a clonal and dominant phenotype in experimental infections. Infection and Immunity 69, 73657373.
Gelanew, T, Hailu, A, Schönian, G, Lewis, MD, Miles, MA and Yeo, M (2014) Multilocus sequence and microsatellite identification of intra-specific hybrids and ancestor-like donors among natural Ethiopian isolates of Leishmania donovani. International Journal of Parasitology 44, 751757.
Inbar, E, Akopyants, NS, Charmoy, M, Romano, A, Lawyer, P, Elnaiem, DE, Kauffmann, F, Barhoumi, M, Grigg, M, Owens, K, Fay, M, Dobson, DE, Shaik, J, Beverley, SM and Sacks, D (2013) The mating competence of geographically diverse Leishmania major strains in their natural and unnatural sand fly vectors. PLoS Genetics 9, e1003672.10.1371/journal.pgen.1003672
Kato, H, Cáceres, AG and Hashiguchi, Y (2016) First evidence of a hybrid of Leishmania (viannia) braziliensis/l. (V.) peruviana DNA detected from the phlebotomine sand fly lutzomyia tejadai in Peru. PLos Neglected Tropical Diseases 10, e0004336.
Keenan, CM, Hendricks, LD, Lightner, L and Johnson, AJ (1984) Visceral leishmaniasis in the German shepherd dog. II. Pathology. Veterinary Pathology 21, 8086.
Lewis, MD, Llewellyn, MS, Yeo, M, Acosta, N, Gaunt, MW and Miles, MA (2011) Recent, independent and anthropogenic origins of Trypanosoma cruzi hybrids. PLoS Neglected Tropical Diseases 5, e1363.
Maia, C, Rolão, N, Nunes, M, Gonçalves, L and Campino, L (2007) Infectivity of five different types of macrophages by Leishmania infantum. Acta Tropica 103, 150155.
Maia, C, Nunes, M, Marques, M, Henriques, S, Rolão, N and Campino, L (2013) In vitro drug susceptibility of Leishmania infantum isolated from humans and dogs. Experimental Parasitology 135, 3641.
Méndez, S, Fernández-Pérez, FJ, Santín, M, De La Fuente, C, Cuquerella, M, Gómez-Muñoz, MT and Alunda, JM (2001) Correlation between in vitro and in vivo infectivity of Leishmania infantum clones. The Journal of Eukaryotic Microbiology 48, 616621.
Miles, MA, Yeo, M and Mauricio, IL (2009) Genetics. Leishmania exploit sex. Science 324, 187189.
Moreira, D, Santarem, N, Loureiro, I, Tavares, J, Silva, AM, Amorim, AM, Ouaissi, A, Cordeiro-da-Silva, A and Silvestre, R (2012) Impact of continuous axenic cultivation in Leishmania infantum virulence. PLoS Neglected Tropical Disease 6, e1469.10.1371/journal.pntd.0001469
Motazedian, MH, Parhizkari, M, Mehrabani, D, Hatam, G and Asgari, Q (2010) First detection of Leishmania major in rattus norvegicus from Fars Province, Southern Iran. Vector Borne and Zoonotic Diseases 10, 969975.10.1089/vbz.2008.0214
Nolder, D, Roncal, N, Davies, CR, Llanos-Cuentas, A and Miles, MA (2007) Multiple hybrid genotypes of Leishmania (viannia) in a focus of Mucocutaneous leishmaniasis. American Society of Tropical Medicine and Hygiene 76, 573578.10.4269/ajtmh.2007.76.573
Odiwuor, S, De Doncker, S, Maes, I, Dujardin, JC and Van der Auwera, G (2011) Natural Leishmania donovani/leishmania aethiopica hybrids identified from Ethiopia. Infection, Genetics and Evolution 11, 21132118.
Ravel, C, Cortes, S, Pratlong, F, Morio, F, Dedet, JP and Campino, L (2006) First report of genetic hybrids between two very divergente Leishmania species: Leishmania infantum and Leishmania major. International Journal for Parasitology 36, 13831388.10.1016/j.ijpara.2006.06.019
Rogers, MB, Downing, T, Smith, BA, Imamura, H, Sanders, M, Svobodova, M, Volf, P, Berriman, M, Cotton, JA and Smith, DF (2014) Genomic confirmation of hybridisation and recent inbreeding in a vector-isolated Leishmania population. PLoS Genetics 10, e1004092.10.1371/journal.pgen.1004092
Romano, A, Inbar, E, Debrabant, A, Charmoy, M, Lawyer, P, Ribeiro-Gomes, F, Barhoumi, M, Grigg, M, Shaik, J, Dobson, D, Beverley, SM and Sacks, DL (2014) Cross-species genetic exchange between visceral and cutaneous strains of Leishmania in the sand fly vector. Proceedings of the National Academy of Sciences of the United States of America 111, 1680816813.
Rougeron, V, De Meeûs, T and Bañuls, AL (2015) A primer for Leishmania population genetic studies. Trends in Parasitology 31, 5259.
Rougeron, V, De Meeûs, T and Bañuls, AL (2017) Reproduction in Leishmania: a focus on genetic exchange. Infection, Genetics and Evolution 50, 128132.
Sadlova, J, Yeo, M, Seblova, V, Lewis, MD, Maurício, I, Volf, P and Miles, MA (2011) Visualisation of Leishmania donovani FluorescentHybrids during early stage development in the Sand FlyVector. PLoS ONE 6, e19851.
Salguero, FJ, Garcia-Jimenez, WL, Lima, I and Seifert, K (2018) Histopathological and immunohistochemical characterisation of hepatic granulomas in Leishmania donovani-infected BALB/c mice: a time-course study. Parasites & Vectors 11, 73.
Santana, CC, Vassallo, J, de Freitas, LA, Oliveira, GG, Pontes-de-Carvalho, LC and dos-Santos, WL (2008) Inflammation and structural changes of splenic lymphoid tissue in visceral leishmaniasis: a study on naturally infected dogs. Parasite Immunology 30, 515524.
Schönian, G, Nasereddin, A, Dinse, N, Schweynoch, C, Schallig, HD, Presber, W and Jaffe, CL (2003) PCR diagnosis and characterization of Leishmania in local and imported clinical samples. Diagnostic Microbiology and Infectious Disease 47, 349358.
Silva-O'Hare, J, de Oliveira, IS, Klevorn, T, Almeida, VA, Oliveira, GG, Atta, AM, de Freitas, LA and Dos-Santos, WL (2016) Disruption of splenic lymphoid tissue and plasmacytosis in canine visceral leishmaniasis: changes in homing and survival of plasma cells. PLoS ONE 11, e0156733.
Smelt, SC, Engwerda, CR, McCrossen, M and Kaye, PM (1997) Destruction of follicular dendritic cells during chronic visceral leishmaniasis. Journal of Immunology 158, 38133821.
Tomás-Pérez, M, Khaldi, M, Riera, C, Mozo-León, D, Ribas, A, Hide, M, Barech, G, Benyettou, M, Seghiri, K, Doudou, S and Fisa, R (2014) First report of natural infection in hedgehogs with Leishmania major, a possible reservoir of zoonotic cutaneous leishmaniasis in Algeria. Acta Tropica 135, 4449.
Torrico, MC, De Doncker, S, Arevalo, J, Le Ray, D and Dujardin, JC (1999) In vitro promastigote fitness of putative Leishmania (viannia) braziliensis/leishmania (Viannia) peruviana hybrids. Acta Tropica 72, 99110.
Vanaerschot, M, Maes, I, Ouakad, M, Adaui, V, Maes, L, De Doncker, S, Rijal, S, Chappuis, F, Dujardin, JC and Decuypere, S (2010) Linking in vitro and in vivo survival of clinical Leishmania donovani strains. PLoS ONE 5, e12211.
Veress, B, Abdalla, RE and El Hassan, AM (1983) Visceral spreading depletion of thymus-dependent regions and amyloidosis in mice and hamsters infected intradermally with Leishmania isolated from Sudanese cutaneous leishmaniasis. Brasilian Journal of Experimental Pathology 64, 505514.
Villaseñor-Cardoso, MI, Salaiza, N, Delgado, J, Gutiérrez-Kobeh, L, Pérez-Torres, A and Becker, I (2008) Mast cells are activated by Leishmania mexicana LPG and regulate the disease outcome depending on the genetic background of the host. Parasite Immunology 30, 425434.
Volf, P, Benkova, I, Myskova, J, Sadlova, J, Campino, L and Ravel, C (2007) Increased transmission potential of Leishmania major/leishmania infantum hybrids. International Journal for Parasitology 37, 589593.
WHO (2010) Control of the leishmaniasis: report of a meeting of the WHO Expert Committee on the Control of Leishmaniases. WHO Technical Report Series no. 949. World Health Organization. Geneva, Switzerland.
WHO (2015) Investing to overcome the global impact of neglected tropical diseases: third WHO report on neglected tropical diseases. Third WHO Report on neglected tropical diseases. Geneva, Switzerland.
Zamora-Chimal, J, Fernández-Figueroa, EA, Ruiz-Remigio, A, Wilkins-Rodríguez, AA, Delgado-Domínguez, J, Salaiza-Suazo, N, Gutiérrez-Kobeh, L and Becker, I (2017) NKT cell activation by Leishmania mexicana LPG: description of a novel pathway. Immunobiology 222, 454462.
Zilberstein, D and Shapira, M (1994) The role of pH and temperature in the development of Leishmania parasites. Annual Review of Microbiology 48, 449470.


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Elucidating in vitro and in vivo phenotypic behaviour of L. infantum/L. major natural hybrids

  • S. Cortes (a1), A. Albuquerque-Wendt (a1), C. Maia (a1), M. Carvalho (a1) (a2), I. A. Lima (a3), L. A. R. de Freitas (a3), W. L. C. dos-Santos (a3) and L. Campino (a1)...


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