Hostname: page-component-586b7cd67f-t7czq Total loading time: 0 Render date: 2024-12-01T03:05:00.276Z Has data issue: false hasContentIssue false
  • English
  • Français

Metacyclogenesis of Leishmania (Viannia) guyanensis: a comprehensive study of the main transformation features in axenic culture and purification of metacyclic promastigotes by negative selection with Bauhinia purpurea lectin

Published online by Cambridge University Press:  27 December 2018

Bárbara Pinheiro Mendes ,
Ildefonso Alves da Silva Jr. Open the ORCID record for Ildefonso Alves da Silva Jr. [Opens in a new window] ,
Jarina Pena DaMata ,
Thiago Castro-Gomes Open the ORCID record for Thiago Castro-Gomes [Opens in a new window] ,
Leda Quercia Vieira Open the ORCID record for Leda Quercia Vieira [Opens in a new window] ,
Fátima Ribeiro-Dias Open the ORCID record for Fátima Ribeiro-Dias [Opens in a new window]  and
Maria Fátima Horta Open the ORCID record for Maria Fátima Horta [Opens in a new window]
Bárbara Pinheiro Mendes
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Ildefonso Alves da Silva Jr.
Affiliation:
Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goias, 74605-050, Goiânia, GO, Brazil
Jarina Pena DaMata
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Thiago Castro-Gomes
Affiliation:
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Leda Quercia Vieira
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Fátima Ribeiro-Dias
Affiliation:
Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goias, 74605-050, Goiânia, GO, Brazil
Maria Fátima Horta*
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
*
Author for correspondence: M. Fátima Horta, E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Leishmania (Viannia) guyanensis is one species that causes cutaneous leishmaniasis in the New World. The incidence of infections with this parasite is probably underestimated and few studies exist on this species, despite its epidemiological importance. In particular, there are no studies concerning L. guyanensis metacyclogenesis and no technique for obtaining metacyclic promastigotes for this species is presently available. Here, we have studied L. guyanensis metacyclogenesis in axenic culture, describing the main changes that occur during this process, namely, in morphology and size, sensitivity to complement-mediated lysis, surface carbohydrates and infectivity to macrophages. We have shown that metacyclogenesis in L. guyanensis promastigotes is basically complete on the 4th day of culture, as determined by decreased body size, increased flagellum length, resistance to complement-mediated lysis and infectivity. We have also found that only a fraction of the parasites is agglutinated by Bauhinia purpurea lectin. The non-agglutinated parasites, which also peaked on the 4th day of culture, had all morphological traits typical of the metacyclic stage. This is the first report describing metacyclogenesis in L. guyanensis axenic promastigotes and a simple and efficient method for the purification of metacyclic forms. Furthermore, a model of human macrophage infection with L. guyanensis was established.

Keywords

Bauhinia purpureaLeishmania guyanensismetacyclogenesis
Type
Research Article
Information
Parasitology , Volume 146 , Issue 6 , May 2019 , pp. 716 - 727
Check for updates
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Almeida, MC, Cuba, CA, de Sá, 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, 325–329.Google Scholar
Almeida, MC, Vilhena, V, Barral, A and Barral-Netto, M (2003) Leishmanial infection: analysis of its first steps: a review. Memórias do Instituto Oswaldo Cruz 98, 861–870.Google Scholar
Bates, PA (2008) Leishmania sand fly interaction: progress and challenges. Current Opinion in Microbiology 11, 340–344.Google Scholar
Bogdan, C, Gessner, A, Solbach, W and Röllinghoff, M (1996) Invasion, control and persistence of Leishmania parasites. Current Opinion in Immunology 8, 517–525.Google Scholar
Brittingham, A, Morrison, CJ, McMaster, WR, McGwire, BS, Chang, KP and Mosser, DM (1995) Role of the Leishmania surface protease gp63 in complement fixation, cell adhesion, and resistance to complement-mediated lysis. Journal of Immunology 155, 3102–3111.Google Scholar
Brodin, TN, Heath, S and Sacks, DL (1992) Genes selectively expressed in the infectious (metacyclic) stage of Leishmania major promastigotes encode a potential basic-zipper structural motif. Molecular and Biochemical Parasitology 52, 241–250.Google Scholar
Camus, D, Zalis, MG, Vannier-Santos, MA and Banic, DM (1995) The art of parasite survival. Brazilian Journal of Medical and Biological Research 28, 399–413.Google Scholar
Chaves, CS, Soares, DC, Silva, RP and Saraiva, EM (2003) Characterization of the species- and stage-specificity of two monoclonal antibodies against Leishmania amazonensis. Experimental Parasitology 103, 152–159.Google Scholar
Courret, N, Prina, E, Mougneau, E, Saraiva, EM, Sacks, DL, Glaichenhaus, N and Antoine, JC (1999) Presentation of the Leishmania antigen LACK by infected macrophages is dependent upon the virulence of the phagocytosed parasites. European Journal of Immunology 29, 762–773.Google Scholar
Cunningham, ML, Titus, RG, Turco, SJ and Beverley, SM (2001) Regulation of differentiation to the infective stage of the protozoan parasite Leishmania major by tetrahydrobiopterin. Science 292, 285–287.Google Scholar
Da Silva, R and Sacks, DL (1987) Metacyclogenesis is a major determinant of Leishmania promastigote virulence and attenuation. Infection and Immunity 55, 2802–2806.Google Scholar
Da Silva, IA Jr, Morato, CI, Quixabeira, VB, Pereira, LI, Dorta, ML, de Oliveira, MA, Horta, MF and Ribeiro-Dias, F (2015) In vitro metacyclogenesis of Leishmania (viannia) braziliensis and Leishmania (leishmania) amazonensis clinical field isolates, as evaluated by morphology, complement-resistance, and infectivity to human macrophages. Biomedical Research International 2015, 393049.Google Scholar
Dutta, A, Bandyopadhyay, S, Mandal, C and Chatterjee, M (2005) Development of a modified MTT assay for screening antimonial resistant field isolates of Indian visceral leishmaniasis. Parasitology International 54, 119–122.Google Scholar
Franke, ED, McGreevy, PB, Katz, SP and Sacks, DL (1985) Growth cycle-dependent generation of complement-resistant Leishmania promastigotes. Journal of Immunology 134, 2713–2718.Google Scholar
Gamboa, D, Torres, K, De Doncker, S, Zimic, M, Arevalo, J and Dujardin, JC (2008) Evaluation of an in vitro and in vivo model for experimental infection with Leishmania (viannia) braziliensis and L. (V.) peruviana. Parasitology 135, 319–326.Google Scholar
Gazola, KC, Ferreira, AV, Anacleto, C, Michalick, MS, Andrade, AF and Moreira, ES (2001) Cell surface carbohydrates and in vivo infectivity of glucantime-sensitive and resistant Leishmania (viannia) guyanensis cell lines. Parasitology Research 87, 935–940.Google Scholar
Gaur, U, Showalter, M, Hickerson, S, Dalvi, R, Turco, SJ, Wilson, ME and Beverley, SM (2009) Leishmania donovani lacking the Golgi GDP-Man transporter LPG2 exhibit attenuated virulence in mammalian hosts. Experimental Parasitology 122, 182–191.Google Scholar
Giannini, MS (1974) Effects of promastigote growth phase, frequency of subculture, and host age on promastigote-initiated infections with Leishmania donovani in the golden hamster. Journal of Protozoology 21, 521–527.Google Scholar
Howard, MK, Sayers, G and Miles, MA (1987) Leishmania donovani metacyclic promastigotes: transformation in vitro, lectin agglutination, complement-resistance, and infectivity. Experimental Parasitology 64, 147–156.Google Scholar
Jennings, YL, de Souza, AA, Ishikawa, EA, Shaw, JJ, Lainson, R and Silveira, F (2014) Phenotypic characterization of Leishmania spp. causing cutaneous leishmaniasis in the lower Amazon region, western Pará state, Brasil, reveals a putative hybrid parasite, Leishmania (viannia) guyanensis x Leishmania (viannia) shawi shawi. Parasite 21, 39.Google Scholar
Jokiranta, TS, Jokipii, L and Meri, S (1995) Complement-resistance of parasites. Scandinavian Journal of Immunology 42, 9–20.Google Scholar
Lainson, R, Shaw, JJ, Miles, MA and Póvoa, M (1982) Leishmaniasis in Brazil: XVII enzymic characterization of a Leishmania from the armadillo, Dasypus novemcinctus (Edentata), from Pará State. Transactions of the Royal Society of Tropical Medicine and Hygiene 76, 810–811.Google Scholar
Lincoln, LM, Ozaki, M, Donelson, JE and Beetham, JK (2004) Genetic complementation of Leishmania deficient in PSA (GP46) restores their resistance to lysis by complement. Molecular and Biochemical Parasitology 137, 185–189.Google Scholar
Lira, R, Méndez, S, Carrera, L, Jaffe, C, Neva, F and Sacks, DL (1998) Leishmania tropica: the identification and purification of metacyclic promastigotes and use in establishing mouse and hamster models of cutaneous and visceral disease. Experimental Parasitology 89, 331–342.Google Scholar
Louassini, M, Adroher, FJ, Foulquié, MR and Benítez, R (1998) Investigations on the in vitro metacyclogenesis of a visceral and a cutaneous human strain of Leishmania infantum. Acta Tropica 70, 355–368.Google Scholar
Mahoney, AB, Sacks, DL, Saraiva, E, Modi, G and Turco, SJ (1999) Intra-species and stage-specific polymorphisms in lipophosphoglycan structure control Leishmania donovani-sand fly interactions. Biochemistry 38, 9813–9823.Google Scholar
Marín, M, Muskus, C, Ramírez, JR, Arbelaez, LF, Alzate, JF and Berberich, C (2000) The gene encoding the metacyclogenesis-associated transcript Mat-1 is conserved in the genus Leishmania and shows a tendency to form dimmers upon protein expression. Parasitology Research 86, 431–435.Google Scholar
McConville, MJ, Turco, SJ, Ferguson, MA and Sacks, DL (1992) Developmental modification of lipophosphoglycan during the differentiation of Leishmania major promastigotes to an infectious stage. EMBO Journal 11, 3593–3600.Google Scholar
McKean, PG, Denny, PW, Knuepfer, E, Keen, JK and Smith, DF (2001) Phenotypic changes associated with deletion and overexpression of a stage-regulated gene family in Leishmania. Cellular Microbiology 3, 511–523.Google Scholar
Mosser, DM and Brittingham, A (1997) Leishmania, macrophages and complement: a tale of subversion and exploitation. Parasitology 115, 9–23.Google Scholar
Muskus, C, Segura, I, Oddone, R, Turco, SJ, Leiby, DA, Toro, L, Robledos, S and Saravia, NG (1997) Carbohydrate and LPG expression in Leishmania viannia subgenus. Journal of Parasitology 83, 671–678.Google Scholar
Nieves, E and Pimenta, PFP (2000) Development of Leishmania (viannia) braziliensis and Leishmania (leishmania) amazonensis in the sand fly Lutzomyia migonei (Diptera: Psychodidae). Journal of Medical Entomology 37, 134–140.Google Scholar
Pinto-da-Silva, LH, Camurate, M, Costa, KA, Oliveira, SM, Da Cunha-e-Silva, NL and Saraiva, EM (2002) Leishmania (Viannia) braziliensis metacyclic promastigotes purified using Bauhinia purpurea lectin are complement-resistant and highly infective form macrophages in vitro and hamsters in vivo. International Journal for Parasitology 32, 1371–1377.Google Scholar
Pinto-da-Silva, LH, Fampa, P, Soares, DC, Oliveira, SM, Souto-Padron, T and Saraiva, EM (2005) The 3A1-La monoclonal antibody reveals key features of Leishmania (l) amazonensis metacyclic promastigotes and inhibits procyclics attachment to the sand fly midgut. International Journal for Parasitology 35, 757–764.Google Scholar
Pires, AS, Borges, AF, Cappellazzo-Coelho, A, Dorta, ML, Lino-Junior, R de S, Pereira, LI, Pinto, SA, Pelli-de-Oliveira, MA, de Matos, GG, Abrahamsohn, IA, Uliana, SR, Lima, GM and Ribeiro-Dias, F (2015) Identification and biological characterization of Leishmania (viannia) guyanensis isolated from a patient with tegumentary leishmaniasis in Goiás, a no endemic area for this species in Brazil. Biomedical Research International 2015, 350764.Google Scholar
PubMed. https://www.ncbi.nlm.nih.gov/pubmed (Accessed 8 May 2018).Google Scholar
Puentes, SM, Sacks, DL, Silva, RP and Joiner, KA (1988) Complement biding by two developmental stages of Leishmania major promastigotes varying in expression of a surface lipophosphoglycan. Journal of Experimental Medicine 167, 887–902.Google Scholar
Ramos, CS, Franco, FAL, Smith, DF and Uliana, SRB (2004) Characterization of a new Leishmania META gene and genomic analysis of the META cluster. FEMS Microbiology Letters 238, 213–219.Google Scholar
Ramos, CS, Yokoyama-Yasunaka, JKU, Guerra-Giraldez, C, Price, HP, Mortara, RA, Smith, DF and Uliana, SRB (2011) Leishmania amazonensis META2 protein confers protection against heat shock and oxidative stress. Experimental Parasitology 127, 228–237.Google Scholar
Sacks, DL (1989) Metacyclogenesis in Leishmania promastigotes. Experimental Parasitology 69, 100–103.Google Scholar
Sacks, DL and Da Silva, RP (1987) The generation of infective stage Leishmania major promastigotes is associated with the cell surface expression and release of a developmentally regulated glycolipid. Journal of Immunology 139, 3099–3106.Google Scholar
Sacks, DL and Perkins, PV (1984) Identification of an infective stage of Leishmania promastigotes. Science 223, 1417–1419.Google Scholar
Sacks, DL and Perkins, PV (1985) Development of infective stage Leishmania promastigotes within phlebotomine sandflies. American Journal of Tropical Medicine and Hygiene 34, 456–459.Google Scholar
Sacks, DL, Brodin, TN and Turco, SJ (1990) Developmental modification of the lipophosphoglycan from Leishmania major promastigotes during metacyclogenesis. Molecular and Biochemical Parasitology 42, 225–233.Google Scholar
Sacks, DL, Pimenta, PF, McConville, MJ, Schneider, P and Turco, SJ (1995) Stage-specific binding of Leishmania donovani to the sand fly vector midgut is regulated by conformational changes in the abundant surface lipophosphoglycan. Journal of Experimental Medicine 181, 685–697.Google Scholar
Sádlová, J, Price, HP, Smith, BA, Votýpka, J, Volf, P and Smith, DF (2000) The stage-regulated HASPB and SHERP proteins are essential for differentiation of the protozoan parasite Leishmania major in its sand fly vector, Phlebotomus papatasi. Cellular Microbiology 12, 1765–1779.Google Scholar
Santos, MG, Silva, MF, Zampieri, RA, Lafraia, RM and Floeter-Winte, LM (2011) Correlation of meta 1 expression with culture stage, cell morphology and infectivity in Leishmania (leishmania) amazonensis. Memórias do Instituto Oswaldo Cruz 106, 190–193.Google Scholar
Saraiva, EM, Pimenta, PF, Brodin, TN, Rowton, E, Modi, GB and Sacks, DL (1995) Changes in lipophosphoglycan and gene expression associated with the development of Leishmania major in Phlebotomus papatasi. Parasitology 111, 275–287.Google Scholar
Saraiva, EM, Pinto-da-Silva, LH, Wanderley, JLM, Bonomo, AC, Barcinski, MA and Moreira, MEC (2005) Flow cytometric assessment of Leishmania spp metacyclic differentiation: validation by morphological features and specific markers. Experimental Parasitology 110, 39–47.Google Scholar
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, 349–358.Google Scholar
Serafim, TD, Figueiredo, AB, Costa, PAC, Marques-da-Silva, EA, Gonçalves, R, de Moura, SA, Gontijo, NF, da Silva, SM, Michakick, MS, Meyer-Fernandes, JR, de Carvalho, RP, Uliana, SR, Fietto, JL and Afonso, LC (2012) Leishmania metacyclogenesis is promoted in the absence of purines. PLoS Neglected Tropical Diseases 6, e1833.Google Scholar
Soares, RP, Cardoso, TL, Barron, T, Araujo, MS and Turco, SJ (2005) Leishmania braziliensis: a novel mechanism in the lipophosphoglycan regulation during metacyclogenesis. International Journal for Parasitology 35, 245–253.Google Scholar
Sousa-Franco, J, Araújo-Mendes, E, Silva-Jardim, I, Lima-Santos, J, Faria, DR, Dutra, WO and Horta, MF (2006) Infection-induced respiratory burst in BALB/c macrophages kills Leishmania guyanensis amastigotes through apoptosis: possible involvement in resistance to cutaneous leishmaniasis. Microbes and Infection 8, 390–400.Google Scholar
Späth, GF and Beverley, SM (2001) A lipophosphoglycan-independent method for isolation of infective Leishmania metacyclic promastigotes by density gradient centrifugation. Experimental Parasitology 99, 97–103.Google Scholar
Späth, GF, Garraway, LA, Turco, SJ and Beverley, SM (2003) The role(s) of lipophosphoglycan (LPG) in the establishment of Leishmania major infections in mammalian hosts. Proceedings of the National Academy of Sciences USA 100, 9536–9541.Google Scholar
Turco, SJ and Descoteaux, A (1992) The lipophosphoglycan of Leishmania parasites. Annual Review of Microbiology 46, 65–94.Google Scholar
Uliana, SRB, Goyal, N, Freymüller, E and Smith, DF (1999) Leishmania: overexpression and comparative structural analysis of the stage-regulated meta 1 gene. Experimental Parasitology 92, 1983–1191.Google Scholar
Yao, C, Chen, Y, Sudan, B, Donelson, JE and Wilson, ME (2008) Leishmania chagasi: homogenous metacyclic promastigotes isolated by buoyant density are highly virulent in a mouse model. Experimental Parasitology 118, 129–133.Google Scholar
Zakai, HA, Chance, ML and Bates, PA (1998) In vitro stimulation of metacyclogenesis in Leishmania braziliensis, L. donovani, L. major and L. mexicana. Parasitology 116, 305–309.Google Scholar