Single Cell Study of Malaria and Antimalarial Drug Impacts on Platelets and Mast Cells

Abstract

Platelets and mast cells are among the first responders against foreign substances within a biological system. Understanding how the fundamental mechanisms of these two cell types act in a diseased vs. healthy environment is particularly important for any therapeutic approach. By using an electrochemical single cell technique, carbon-fiber microelectrode amperometry (CFMA), platelets and mast cells were studied during malaria infection. Lastly, platelets were studied for their response to two common antimalarial drugs using the same single cell technique. Chapter 1 gives an overview of single cell analysis techniques (optical,electrochemical, mass spectrometric, and microfluidic methods) and their recent progress in biological applications. Single cell analysis techniques allow not only for quantification of intracellular biological molecules, but also for characterization of dynamic cellular processes with ultrahigh spatiotemporal resolution. Thus, the obtained information allows for analysis of fundamental mechanisms underlying cellular behavior and function. Chapter 2 focuses on platelets and malaria and established the malaria model that willalso be used for mast cells and malaria studies in Chapter 3. Chapter 2 starts with an emphasis on creating a live malaria-infected mouse model system and establishing two distinct stages of infection (ascending and descending parasitemia). Then, the chapter goes into detail about the use of CFMA to capture information on how platelets from malaria-infected conditions were impacted compared to platelets from malaria-free mice. Chapter 3 focuses on mast cells and malaria and a comparison to platelets data in Chapter 2. The work in this chapter first involves obtaining mast cells from malaria-infected and healthy mice and co-culturing the mast cells with fibroblast cells to achieve successful primary culture for single cell studies. Then, the chapter details the use of CFMA to obtain relevant biological information about mast cells in the context of malaria. Finally, the chapter concludes with a comparison to the information obtained from platelets. Chapter 4 focuses on platelets and their response to two select antimalarial drugs.The selected drugs are commonly used drugs to treat malarial infection by targeting the lifecycle of the malarial parasites. The work in this chapter involves using CFMA and a bulk electrochemical technique to study the effect of chloroquine and quinine on platelets and to obtain relevant biological information about platelet performance during malaria treatment. In all, CFMA as a single cell technique was able to reveal that platelets and mast cells were affected at the fundamental level in the context of malaria and antimalarial drugs.