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Background:    Leishmania major  and  Leishmania tropica  are the main causative agents of cutaneous leishmaniasis. Proteomics as a novel approaches could be used to evaluate protein expression levels in different stages of  Leishmania  species. We compare the protein contents of amastigote-like forms in  L. tropica  and  L. major  using two-dimensional gel electrophoresis (2-DE) and bioinformatics methods.Materials and Methods:   Leishmania  parasites were isolated from the lesions of Iranian patients and identified using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). Five isolates of each two species were cultured in specific media to obtain amastigote-like forms to be prepared for proteomics study. Total protein contents were separated using 2-DE. The gels were stained by silver nitrate and scan was imaged. The protein spots with different expression changes in each gel were analyzed using Progenesis SameSpots software.Results:   A total of 354 protein spots were detected in both amastigote-like forms. Comparative analysis of protein spots with different expressions in the two amastigote-like form species showed 173 highly expressed spots of which 74  L. tropica  and 99  L. major  proteins were spotted with fold≥2. Also, 16 and 20 new protein spots were uniquely found in  L. tropica  and  L. major , respectively. Clustering of different detected proteins using correlation analysis divided the proteins into two clusters based on their expression level. Furthermore, clustering results were confirmed by principal component analysis.Conclusion:   Using proteomics methods specially 2-DE and statistical analysis demonstrated significant changes in protein expression levels in amastigote-like forms of  L. tropica  and  L. major  isolates.

Comparative Two-dimensional Gel Electrophoresis Maps for Amastigote-like Proteomes of Iranian Leishmania Tropica and Leishmania major Isolates