title: A Leishmania (L.) amazonensis ATP diphosphohydrolase isoform and potato apyrase share epitopes: antigenicity and correlation with disease progression.
abstract: A Leishmania (Leishmania) amazonensis ATP diphosphohydrolase isoform was partially purified from plasma membrane of promastigotes by preparative non-denaturing polyacrylamide gel electrophoresis. SDS-PAGE followed by Western blots developed with polyclonal anti-potato apyrase antibodies identified diffuse bands of about 58-63 kDa, possibly glycosylated forms of this protein. By ELISA technique, a significantly higher total IgG antibody level against potato apyrase was found in serum from promastigote-infected mice, as compared to the uninfected mice, confirming both the existence of shared epitopes between the parasite and vegetable proteins, and the parasite ATP diphosphohydrolase antigenicity. By Western blotting, serum from amastigote-infected BALB/c mice recognizes both potato apyrase and this antigenic ATP diphosphohydrolase isoform isolated from promastigotes, suggesting that it is also expressed in the amastigote stage. The infection monitored along a 90-day period in amastigote-infected mice showed reactivity of IgG2a antibody in early steps of infection, while the disappearance of the IgG2a response and elevation of IgG1 antibody serum levels against that shared epitopes were associated with the progression of experimental leishmaniasis. This is the first observation of the antigenicity of a L. (L.) amazonensis ATP diphosphohydrolase isoform, and of the ability of cross-immunoreactivity with potato apyrase to differentiate serologically stages of leishmaniasis in infected mice.
authors.affiliation: Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil.
authors.forename: E S
authors.initials: ES
authors.lastname: Coimbra
authors.forename: S C
authors.initials: SC
authors.lastname: Gonçalves-da-Costa
authors.forename: B L S
authors.initials: BL
authors.lastname: Costa
authors.forename: N L L
authors.initials: NL
authors.lastname: Giarola
authors.forename: F A
authors.initials: FA
authors.lastname: Rezende-Soares
authors.forename: M R
authors.initials: MR
authors.lastname: Fessel
authors.forename: A P
authors.initials: AP
authors.lastname: Ferreira
authors.forename: C S F
authors.initials: CS
authors.lastname: Souza
authors.forename: A L
authors.initials: AL
authors.lastname: Abreu-Silva
authors.forename: E G
authors.initials: EG
authors.lastname: Vasconcelos
mesh_terms: D000818:Animals; D000940:Antigenic Variation; D001081:Apyrase / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western; D003429:Cross Reactions; D018450:Disease Progression; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005260:Female; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D007894:Leishmania mexicana / Q000201:enzymology*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D051379:Mice; D008807:Mice, Inbred BALB C; D011198:Solanum tuberosum / Q000201:enzymology*
chemical_list: D000939:Epitopes; D007527:Isoenzymes; D001081:Apyrase