Fig 1.
Experimental design and tracking of ASC tdTomato+.
Panel A shows the experimental design of the study. C57BL/6 mice were infected with S. mansoni (Day 0), treated orally with PZQ (Day 45; PZQ group), and then injected with ASC (3x105) (Day 75; PZQ/ASC group) or PBS (PZQ). The mice from PZQ and PZQ/ASC groups were analyzed on Day 90. In parallel, other group treated with PZQ was injected with 3x105 tdTomato-expressing ASC derived from ROSAmT/mG mice. After 1 day, 7 days, and 15 days, the animals were euthanized, and their liver and lungs collected for confocal analysis. Note: an infected not treated group (NT group) was followed until day 90-post infection as a control for the PZQ treatment. Panel B shows representative confocal cell images of the lung and the liver 1, 7 and 15 days after ASCtdTomato+ injection. Cryosections were counterstained with DAPI (blue fluorescence). Red fluorescence refers to ASCtdTomato+. Images were captured using the Ti-E C2 plus microscope confocal with magnification of 100x, and analyzed using NIS-Elements (Nikon).
Fig 2.
ASC effect in S. mansoni infection.
Panels A-E (mean ± standard deviation) show the liver weight (A), number of helminths recovered from mesenteric vessels (B), percentage of viable eggs (C), number (D) and volume (E) of granulomas in the liver from Schistosoma-infected C57BL/6 mice treated with PZQ coadministered with ASC. Dashed lines represent the mean of non-infected mice. The horizontal lines indicate differences statistically significant (p<0.05) between groups. Representative images of the liver (F), helminths recovered (G), viable/nonviable eggs (H) and granulomas (I) stained with HE. NT = infected not treated group; PZQ = infected treated with praziquantel group; PZQ/ASC = infected treated with praziquantel plus adipose tissue-derived stem cells group.
Fig 3.
ASC therapy effect on granulomas and ALT levels.
Panel A shows a schematic representation of the phases and cellular composition of granulomas. The exudative phase is characterized by the establishment and organization of the granulomatous reaction with focal destruction of the vessel walls involved and/or adjacent parenchymal and abundant presence of eosinophils, macrophages and other immunologic cells. The early phase is characterized by the presence of giant cells and the dissociation of collagen fibers. An increase in collagen characterizes the advanced phase. Fibrosis is characterized by a significant decrease in cellular composition and thickening of the collagen layer. Panel B shows the comparative analysis of the phase and cellular composition of the granulomas in PZQ and PZQ/ASC groups. Panel C shows representative images from the liver granulomas stained with Masson’s trichrome. Panel D shows fibrotic area in the liver. Panel E shows the levels of ALT in sera of Schistosoma-infected C57BL/6 mice treated with PZQ or PZQ/ASC, expressed as the mean ± standard deviation. The horizontal lines indicate statistically significant differences between the groups (p<0.05). PZQ = Praziquantel-treated group. PZQ/ASC = Praziquantel plus adipose tissue-derived stem cells-treated group.
Fig 4.
T cell activation before and after ASC therapy.
Frequency of CD4+ and CD8+ T lymphocytes expressing CD69, CD25, CD28 and CTLA-4 in spleen (A) and liver (B) cells. The data are shown as means ± standard deviation. Horizontal lines indicate statistically significant differences (p<0.05). PZQ = Praziquantel-treated group; PZQ/ASC = Praziquantel plus adipose tissue-derived stem cells-treated group.
Fig 5.
Cytokine profile before and after ASC therapy.
Panel A shows cytokine levels in situ (liver) and systemic (in serum) from Schistosoma-infected C57BL/6 mice with PZQ only or PZQ/ASC treatment. Panel B shows radar charts summarizing the percentage of high cytokine producers of each group in comparison to the global median value of whole universe of data (NI + PZQ + PZQ/ASC) for each cytokine (S2 Fig). When the frequency of high producers was greater than 50% (on a scale of 0–100%), the result was highlighted. The data are shown as means ± standard deviation. The horizontal lines indicate statistically significant differences (p<0.05). PZQ = Praziquantel-treated group; PZQ/ASC = Praziquantel plus adipose tissue-derived stem cells-treated group.