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Promastigote form of  Leishmania,  an intracellular pathogen, delays phagosome maturation and resides inside macrophages. But till date limited study has been done to manipulate the phagosomal machinery of macrophages to restrict  Leishmania  growth. Attenuated  Leishmania  strain exposed RAW 264.7 cells showed a respiratory burst and enhanced production of pro-inflammatory mediators. The augmentation of pro-inflammatory activity is mostly attributed to p38 MAPK and p44/42 MAPK. In our study, these activated macrophages are found to induce phagosome maturation when infected with pathogenic  Leishmania donovani . Increased co-localization of carboxyfluorescein succinimidyl ester labeled pathogenic  L. donovani  with Lysosome was found. Moreover, increased co-localization was observed between pathogenic  L. donovani  and  l ate phagosomal markers  viz.  Rab7, Lysosomal Associated Membrane Protein 1, Cathepsin D, Rab9, and V-ATPase which indicate phagosome maturation. It was also observed that inhibition of V-type ATPase caused significant hindrance in attenuated  Leishmania  induced phagosome maturation. Finally, it was confirmed that p38 MAPK is the key player in acidification and maturation of phagosome in attenuated  Leishmania  strain pre-exposed macrophages. To our knowledge, this study for the first time reported an approach to induce phagosome maturation in  L. donovani  infected macrophages which could potentiate short-term prophylactic response in future.

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Attenuated Leishmania induce pro-inflammatory mediators and influence leishmanicidal activity by p38 MAPK dependent phagosome maturation in Leishmania donovani co-infected macrophages