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In Vivo Infection with Leishmania amazonensis to Evaluate Parasite Virulence in Mice
Published on: February 20, 2020
A Leishmania (L.) amazonensis ATP diphosphohydrolase isoform and potato apyrase share epitopes: antigenicity and
E S Coimbra1, S C Gonçalves-da-Costa, B L S Costa
1Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil.
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.
Insights
Researchers identified a shared antigen between Leishmania (Leishmania) amazonensis ATP diphosphohydrolase and potato apyrase. This cross-reactivity helps differentiate stages of experimental leishmaniasis in mice.
Area of Science:
- Immunology
- Parasitology
- Biochemistry
Background:
- Leishmania (Leishmania) amazonensis is a protozoan parasite causing leishmaniasis.
- ATP diphosphohydrolase is an enzyme involved in parasite survival and virulence.
- Identifying parasite-specific antigens is crucial for diagnostic development.
Purpose of the Study:
- To investigate the antigenicity of a Leishmania (Leishmania) amazonensis ATP diphosphohydrolase isoform.
- To explore cross-reactivity between the parasite enzyme and potato apyrase.
- To assess the potential of this cross-reactivity in diagnosing different stages of leishmaniasis.
Main Methods:
- Partial purification of ATP diphosphohydrolase from Leishmania (Leishmania) amazonensis promastigotes.
- SDS-PAGE and Western blotting using anti-potato apyrase antibodies.
- ELISA and Western blotting with serum from infected and uninfected mice.
- Monitoring antibody responses (IgG2a, IgG1) over a 90-day infection period.
Main Results:
- A ~58-63 kDa ATP diphosphohydrolase isoform was identified, potentially glycosylated.
- Significant IgG antibody levels against potato apyrase in infected mice confirmed shared epitopes.
- Serum from infected mice recognized both potato apyrase and the parasite enzyme.
- Distinct IgG2a and IgG1 antibody profiles correlated with early and late stages of experimental leishmaniasis.
Conclusions:
- The Leishmania (Leishmania) amazonensis ATP diphosphohydrolase isoform is antigenic.
- Cross-immunoreactivity with potato apyrase exists, indicating shared epitopes.
- This cross-reactivity can serologically differentiate stages of experimental leishmaniasis in mice.
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