Leukosorb Project (2016)
Undergraduate: Preston Setzer
Faculty Advisor: Ilona Jaspers
Department: Biology
Elevated secretions of cytokines and chemokines, like IL-8 and IL-6, in the nasal mucosa can be used as biomarkers of innate immune response to pathogens or pollutants. Nasal lavage is the current gold standard to evaluate these biomarkers in clinical settings. However, this sample collection method is lacking in several areas: samples are diluted, samples can be contaminated by blood, storage requires refrigeration or freezing, and reliable, consistent collection can be difficult in pediatric populations. The goal for this study is to validate a new method using leukosorb paper (Pall Corporation) for collection of proteins present in the nasal mucosa. Leukosorb paper is a highly wettable, fibrous matrix designed specifically for leukocyte absorption, which we cut into thin strips. To test the viability of leukosorb strips for cytokine evaluation, we first wanted to determine whether leukosorb strips would absorb cytokines from the nasal passage and variability in protein recovery from repeated leukosorb strip sampling in the nasal passage. To test this we administered about 150 microliters of aerosolized 0.9% saline solution into each nasal passage, inserted leukosorb strips into each nare of 3 human subjects and clamped the nose for one minute. We repeated this process three consecutive times with 4 minute recovery periods in-between sampling on three separate days. Once all samples were collected, we stored them in a -20 degree C freezer, and evaluated cytokine levels via ELISA. Our next step was to determine the variability in protein recovery from leukosorb strips. To do this, we exposed multiple leukosorb strips to a known concentration of IL-8, stored them in a -20 degree C freezer, and evaluated Il-8 levels via ELISA. Preliminary results suggest that leukosorb strips have the ability to extract nasal proteins from human subjects reliably on the first sample. We also determined that Il-8 can be reliably recovered from leukosorb strips. Future directions for t