Scouring the Lungs for Macrophage Gold

Hey funders! It's been a month since my last update, so I thought I would let folks know what has been going on! Over the past four weeks, our small, intrepid group in the Intensive Care Department has been working on optimizing all aspects of our alveolar macrophage isolation protocol. As a reminder, these are the cells that work to scan our lungs for foreign pathogens (i.e. bacteria, virus, fungi), and initiate a response to the pathogen if need be. For those interested in diving head first into the world of the alveolar macrophage, I'd recommend this review.  

The process of isolating the macrophages requires a few steps. First, we inhale an increasingly salty solution using a nebulizer, and coupled with some breathing exercises (which we are still perfecting), we cough up sputum plugs from deep in the lungs. The saline mobilizes cells, mucus, and other components from deep within the alveoli, and with the breathing exercises and coughing, out comes our desired cells! I've completed the induced sputum protocol twice, and there is a certain pioneering thrill to conducting an experiment on yourself. Next, we have to break apart the viscous and decidedly gross sputum plugs. Using a combination of mechanical dissociation and chemical degradation, we create something akin to a single cell solution, which is necessary for our downstream applications, which includes flow cytometry and cell stimulation assays. Flow cytometry allows us to see what sort of cell populations are present in our sample, which help us ensure that we are working with our desired cell population (the alveolar macrophage). I was rather inexperienced with flow cytometry prior to starting this research project, but with the help of the wonderful faculty and other students at the Radboud umc, I have started to gain the skills needed to utilize this tool effectively! Our end goal is to have a pure population of alveolar macrophages, so that we can stimulate these cells with various pathogens. By trending their responses to pathogen (via production of inter-cellular communiques called cytokines), we can learn more about the functionality of the alveolar macrophages in the context of our sepsis model (the human endotoxemia model), and most importantly, in the context of the different pathologies that bring patients to the intensive care units. Hope you've enjoyed!