In vitro growth kinetics, differentiation and morphological characterisation of Tunisian Leishmania infantum parasites

In this study, a negative peanut agglutinin (PNA) selection was used as a marker for promastigote differentiation to compare the in vitro growth and differentiation kinetics of two visceral and two cutaneous Leishmania (Leishmania) infantum parasites. All parasites had different growth and differentiation kinetics. Cultures initiated with PNA+ parasites purified during the early stationary phase (Day 4), when PNA− (non-agglutinating) parasites peaked, yielded a high PNA− percent. Further morphological analysis at this time point showed that 60-86% of PNA+ forms were procyclics, whilst PNA− forms were composed of 53-71% leptomonads. Nectomonads were present both in PNA− and PNA+ promastigote fractions at nearly equivalent proportions, suggesting that they constitute a transition state in the Leishmania development process, with a fraction of them sharing common constituents of the surface coat with procyclics and the other with leptomonads. Obtaining a high density of promastigotes undergoing developmental differentiation may be useful for further molecular and biochemical identification of developmental stage-specific markers.