Proteomics

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Proteomic analysis of T-cruzi experimentally infected hamster cheeck pouch


ABSTRACT: Mast cells (MCs) were implicated in tissue remodelling in hamster models of heart inflammatory diseases. Here we asked whether MCs promote inflammatory neovascularization in hamster cheek pouch (HCP) parasitized by Trypanosoma cruzi, the protozoan that causes Chagas disease. Intravital microscopy (IVM) performed 3 days after inoculation of Dm28c trypomastigote (GFP-TCTs) showed a subtle infiltration of rhodamine-labeled leukocytes. Interestingly, the baseline levels of FITC-dextran leakage (3 dpi) were consistently increased, raising the possibility that intracellular amastigotes may coopt the low-grade inflammation to facilitate the delivery of plasma-borne nutrients during the period in which host cells must metabolically sustain parasite division. Although proangiogenic indexes were only mildly stimulated at 3 dpi, we found a positive correlation with transcription of proinflammatory cytokines (pro-IL- and IFN-). Having established 3 dpi as a timepoint in which vascular remodeling was already measurable, we next compared the proteomic profiles of parasitized HCP with those of internal controls (PBS injected 3 days earlier in the contralateral pouch) we found 87 proteins upregulated, and 17 were downregulated in the parasitized HCP. Minor changes in protein expression were observed in response to PBS injection (2 upregulated and 12 downregulated proteins. Congruent with the increased density of MCs observed in parasitized HCP (histological analysis), the MC protease 1 (chymase) stood out as the most abundant sequence. Interestingly, toluidine blue staining appointed clusters of MCs degranulating at 3 dpi. Noteworthy, two plasma proteins (complement C3 and serum albumin) were also identified as major proteins, further suggesting that leaky capillaries irrigated the HCP at early stages (3 dpi) of infection . Detection of cytokines or classical proangiogenic factors was beyond the sensitivity of our methods.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Trypanosoma Cruzi Mesocricetus Auratus (golden Hamster)

TISSUE(S): Cheek Pouch

SUBMITTER: Proteomics Unit  

LAB HEAD: Fabio Nogueira

PROVIDER: PXD026905 | Pride | 2022-04-04

REPOSITORIES: Pride

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Publications

Sheltered in Stromal Tissue Cells, <i>Trypanosoma cruzi</i> Orchestrates Inflammatory Neovascularization via Activation of the Mast Cell Chymase Pathway.

Vellasco Lucas L   Svensjö Erik E   Bulant Carlos Alberto CA   Blanco Pablo Javier PJ   Nogueira Fábio F   Domont Gilberto G   de Almeida Natália Pinto NP   Nascimento Clarissa Rodrigues CR   Silva-Dos-Santos Danielle D   Carvalho-Pinto Carla Eponina CE   Medei Emiliano Horácio EH   Almeida Igor C IC   Scharfstein Julio J  

Pathogens (Basel, Switzerland) 20220129 2


Microangiopathy may worsen the clinical outcome of Chagas disease. Given the obstacles to investigating the dynamics of inflammation and angiogenesis in heart tissues parasitized by <i>Trypanosoma cruzi</i>, here we used intravital microscopy (IVM) to investigate microcirculatory alterations in the hamster cheek pouch (HCP) infected by green fluorescent protein-expressing <i>T. cruzi</i> (GFP-<i>T. cruzi</i>). IVM performed 3 days post-infection (3 dpi) consistently showed increased baseline lev  ...[more]

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