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Background  The present study analyzed whether or not the  in vitro  cultivation for long periods of time of pre-isolated  Leishmania amazonensis  from lesions of chronically infected BALB/c mice was able to interfere in the parasites' infectivity using  in vivo  and  in vitro  experiments. In addition, the proteins that presented a significant decrease or increase in their protein expression content were identified applying a proteomic approach.    Methodology/Principal Findings  Parasites were cultured  in vitro  for 150 days. Aliquots were collected on the day 0 of culture (R0), as well as after ten (R10; 50 days of culture), twenty (R20; 100 days of culture), and thirty (R30; 150 days of culture) passages, and were used to analyze the parasites'  in vitro  and  in vivo  infectivity, as well as to perform the proteomic approach. Approximately 837, 967, 935, and 872 spots were found in 2-DE gels prepared from R0, R10, R20, and R30 samples, respectively. A total of 37 spots presented a significant decrease in their intensity of expression, whereas a significant increase in protein content during cultivation could be observed for 19 proteins (both cases >2.0 folds). Some of these identified proteins can be described, such as diagnosis and/or vaccine candidates, while others are involved in the infectivity of  Leishmania . It is interesting to note that six proteins, considered hypothetical in  Leishmania , showed a significant decrease in their expression and were also identified.    Conclusions/Significance  The present study contributes to the understanding that the cultivation of parasites over long periods of time may well be related to the possible loss of infectivity of  L. amazonensis . The identified proteins that presented a significant decrease in their expression during cultivation, including the hypothetical, may also be related to this loss of parasites' infectivity, and applied in future studies, including vaccine candidates and/or immunotherapeutic targets against leishmaniasis.

Identification of Differentially Expressed Proteins from Leishmania amazonensis Associated with the Loss of Virulence of the Parasites