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Tissue Engineering

Tissue Engineering - Poster Session B

Poster L18 - Evaluating Leishmania Donovani Infection Efficiency Using MATLAB Image Analysis

Thursday, October 24, 2024
2:45 PM – 3:45 PM EST
Location: Exhibit Hall E, F & G

Introduction:  Visceral Leishmaniasis, a tropical disease caused by the Leishmania donovani (Ld) parasite, affects over 600,000 people annually, causing liver and spleen enlargement in infected hosts. Despite its 95% mortality rate, the disease remains understudied, and the mechanisms of disease progression are currently unknown. Characterization of the liver’s structure before and after infection revealed an increase in extracellular matrix (ECM) remodeling as the disease progresses. These results identified hepatic stellate cells (HSCs), the ECM producing cells of the liver, as possible role players in Ld infection. We hypothesize that HSCs become more activated, thus producing more ECM, as Ld infection progresses.

Materials and

Methods:  To test our hypothesis, we selected the LX-2 hepatic stellate cell line due to its widespread use and cost efficiency relative to other HSC lines. HSC samples were infected with Ld parasites for 24, 48, and 72 hours. HSC activity was assessed at each time point by quantifying the levels of alpha-Smooth Muscle Actin (α-SMA), a protein associated with cell activation. In order to evaluate the success of the infection, we developed a custom MATLAB script to determine the infection rate of our samples.

Results, Conclusions, and Discussions: Our MATLAB script is able to automatically identify parasite and HSC nuclei, and compares their locations to determine infection rates. We saw an initial increase in α-SMA between the uninfected control and the 24 hour sample. However, we saw a decrease in α-SMA between the 24 and 72 hour timepoints, suggesting activity may decrease over the course of the infection . To support the data from this experiment, we will quantify other markers for cell activity, using ELISA and proliferation assays. In all, the results of this study provided us with a better understanding of how Ld affects the liver in infected hosts, and will inform future experiments that investigate the mechanisms of Leishmaniasis progression.

Acknowledgements (Optional):  We thank Peter Kima and Javier Rosero for their contribution to the project