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Leishmania amazonensis  is the causative agent of American cutaneous leishmaniasis, an important neglected tropical disease. Once  Leishmania amazonensis  is inoculated into the human host, promastigotes are exposed to the extracellular matrix (ECM) of the dermis. However, little is known about the interaction between the ECM and  Leishmania  promastigotes. In this study we established  L. amazonensis  promastigote culture in a three-dimensional (3D) environment mainly composed of Collagen I (COL I). This 3D culture recreates  in vitro  some aspects of the human host infection site, enabling the study of the interaction mechanisms of  L. amazonensis  with the host ECM. Promastigotes exhibited “freeze and run” migration in the 3D COL I matrix, which is completely different from the conventional  in vitro  swimming mode of migration. Moreover,  L. amazonensis  promastigotes were able to invade, migrate inside, and remodel the 3D COL I matrix. Promastigote trans-matrix invasion and the freeze and run migration mode were also observed when macrophages were present in the matrix. At least two classes of proteases, metallo- and cysteine proteases, are involved in the 3D COL I matrix degradation caused by  Leishmania . Treatment with a mixture of protease inhibitors significantly reduced promastigote invasion and migration through this matrix. Together our results demonstrate that  L. amazonensis  promastigotes release proteases and actively remodel their 3D environment, facilitating their migration. This raises the possibility that promastigotes actively interact with their 3D environment during the search for their cellular “home”—macrophages. Supporting this hypothesis, promastigotes migrated faster than macrophages in a novel 3D co-culture model.

peerj

<i>Leishmania amazonensis</i>promastigotes in 3D Collagen I culture: an<i>in vitro</i>physiological environment for the study of extracellular matrix and host cell interactions

Leishmania amazonensispromastigotes in 3D Collagen I culture: anin vitrophysiological environment for the study of extracellular matrix and host cell interactions