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Active tachyzoite migration through 3D collagen I/fibronectin matrix and expansion of its infection in human intestinal MPS generated with Caco-2 double-lumens.

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posted on 2025-02-04, 18:40 authored by Carlos J. Ramírez-Flores, Nicole D. Hryckowian, Andrew N. Gale, Kehinde Adebayo Babatunde, Marcos Lares, David J. Beebe, Sheena C. Kerr, Laura J. Knoll

(A) Schematic representation of bradyzoite/tachyzoite infection niche in human intestinal MPS. (B) Fluorescent images showing the coexistence of EGS LDH2p-GFP/SAG1p-mCherry bradyzoites and tachyzoites in the infected intestinal lumen at 3 days post-infection, with insets highlighting the localization of bradyzoites in the 3D collagen I/fibronectin matrix. Fluorescent images tracking the migration in collagen and Caco-2 infection by EGS LDH2p-GFP/SAG1p-mCherry parasites after exteriorization from Caco-2 cells in the bottom Caco-2 lumen shown in (C) and (D). (C) Micrographs show tachyzoites localizing in the 3D collagen I/fibronectin matrix and actively migrating towards the top Caco-2 lumen by day 5 post-infection. (D) Right micrographs show tachyzoites expanding the infection to the adjacent top Caco-2 lumen. Punctuated circles show parasitic infection to the top Caco-2 lumen. Punctuated lines in white represent the edges of the lumen and its boundary with the matrix. Lumens were infected with 5 × 104 bradyzoites of the EGS LDH2p-GFP/SAG1p-mCherry strain. Magnified photograph in (B) shows different parasites captured in different planes than those in unmagnified photograph. Prominent areas in (C) and (D) are the result from foci of infection. Schematic representation in (A) was created with BioRender.com.

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