title: Leishmania enriettii: biochemical characterisation of lipophosphoglycans (LPGs) and glycoinositolphospholipids (GIPLs) and infectivity to Cavia porcellus.
abstract: BACKGROUND
  Leishmania enriettii is a species non-infectious to man, whose reservoir is the guinea pig Cavia porcellus. Many aspects of the parasite-host interaction in this model are unknown, especially those involving parasite surface molecules. While lipophosphoglycans (LPGs) and glycoinositolphospholipids (GIPLs) of Leishmania species from the Old and New World have already been described, glycoconjugates of L. enriettii and their importance are still unknown.
  
  
  METHODS
  Mice peritoneal macrophages from C57BL/6 and knock-out (TLR2 -/-, TLR4 -/-) were primed with IFN-γ and stimulated with purified LPG and GIPLs from both species. Nitric oxide and cytokine production were performed. MAPKs (p38 and JNK) and NF-kB activation were evaluated in J774.1 macrophages and CHO cells, respectively.
  
  
  RESULTS
  LPGs were extracted, purified and analysed by western-blot, showing that LPG from L88 strain was longer than that of Cobaia strain. LPGs and GIPLs were depolymerised and their sugar content was determined. LPGs from both strains did not present side chains, having the common disaccharide Gal(β1,4)Man(α1)-PO4. The GIPL from L88 strain presented galactose in its structure, suggestive of type II GIPL. On the other hand, the GIPL of Cobaia strain presented an abundance of glucose, a characteristic not previously observed. Mice peritoneal macrophages from C57BL/6 and knock-outs (TLR2 -/- and TLR4 -/-) were primed with IFN-γ and stimulated with glycoconjugates and live parasites. No activation of NO or cytokines was observed with live parasites. On the other hand, LPGs and GIPLs were able to activate the production of NO, IL-6, IL-12 and TNF-α preferably via TRL2. However, in CHO cells, only GIPLs were able to activate TRL2 and TRL4. In vivo studies using male guinea pigs (Cavia porcellus) showed that only strain L88 was able to develop more severe ulcerated lesions especially in the presence of salivary gland extract (SGE).
  
  
  CONCLUSIONS
  The two L. enriettii strains exhibited polymorphisms in their LPGs and GIPLs and those features may be related to a more pro-inflammatory profile in the L88 strain.
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. larissafparanaiba@gmail.com.|Laboratório de Fisiologia de Insetos Hematófagos, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil. larissafparanaiba@gmail.com.
authors.forename: Larissa Ferreira
authors.initials: LF
authors.lastname: Paranaíba
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. rafael.assis@cpqrr.fiocruz.br.
authors.forename: Rafael Ramiro
authors.initials: RR
authors.lastname: de Assis
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. paulamn@cpqrr.fiocruz.br.|Laboratory of Cellular and Molecular Parasitology, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Av. Augusto de Lima 1715, Belo Horizonte, Minas Gerais, 30190-002, Brazil. paulamn@cpqrr.fiocruz.br.
authors.forename: Paula Monalisa
authors.initials: PM
authors.lastname: Nogueira
authors.affiliation: Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários, Departamento de Ciências Biológicas, Campus Diadema, Universidade Federal de São Paulo, UNIFESP, São Paulo, SP, Brazil. ana.trocoli@gmail.com.
authors.forename: Ana Claúdia
authors.initials: AC
authors.lastname: Torrecilhas
authors.affiliation: Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários, Departamento de Ciências Biológicas, Campus Diadema, Universidade Federal de São Paulo, UNIFESP, São Paulo, SP, Brazil. joao.heima@gmail.com.
authors.forename: João Henrique
authors.initials: JH
authors.lastname: Campos
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. amandacardoso@cpqrr.fiocruz.br.
authors.forename: Amanda Cardoso de Oliveira
authors.initials: AC
authors.lastname: Silveira
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. oamfilho@cpqrr.fiocruz.br.
authors.forename: Olindo Assis
authors.initials: OA
authors.lastname: Martins-Filho
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. natalinha_lima@hotmail.com.
authors.forename: Natalia Lima
authors.initials: NL
authors.lastname: Pessoa
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. marcoasc@cpqrr.fiocruz.br.
authors.forename: Marco Antônio
authors.initials: MA
authors.lastname: Campos
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. parreiras@cpqrr.fiocruz.br.
authors.forename: Patrícia Martins
authors.initials: PM
authors.lastname: Parreiras
authors.affiliation: Laboratório de Biologia de Leishmania, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil. normamello@gmail.com.
authors.forename: Maria Norma
authors.initials: MN
authors.lastname: Melo
authors.affiliation: Laboratório de Fisiologia de Insetos Hematófagos, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil. nelder@icb.ufmg.br.
authors.forename: Nelder de Figueiredo
authors.initials: Nde F
authors.lastname: Gontijo
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. rsoares@cpqrr.fiocruz.br.|Laboratory of Cellular and Molecular Parasitology, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, Av. Augusto de Lima 1715, Belo Horizonte, Minas Gerais, 30190-002, Brazil. rsoares@cpqrr.fiocruz.br.
authors.forename: Rodrigo Pedro Pinto
authors.initials: RP
authors.lastname: Soares
mesh_terms: D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D004197:Disease Reservoirs; D006017:Glycolipids / Q000378:metabolism*; D006028:Glycosphingolipids / Q000378:metabolism*; D006168:Guinea Pigs; D018313:Leishmania enriettii / Q000502:physiology*; D007896:Leishmaniasis / Q000469:parasitology*; D017737:Macrophages, Peritoneal / Q000469:parasitology; D008297:Male; D051379:Mice; D009569:Nitric Oxide; D010743:Phospholipids / Q000378:metabolism*; D011576:Psychodidae / Q000469:parasitology
chemical_list: D006017:Glycolipids; D006028:Glycosphingolipids; D010743:Phospholipids; C008290:lipophosphonoglycan; D009569:Nitric Oxide