Soluble factor signaling between immune cells and fibroblasts is critical in regulating biological processes. However, it is often dysregulated in diseases and leads to physiological changes, including airway inflammation in asthma and allergies. One immune cell type that can be attributed to airway inflammation is eosinophils (EOS). When activated by interleukin-3 and heat-aggregated immunoglobulin G, EOS release certain soluble factors associated with the activation of lung fibroblasts. To investigate the interactions between human lung fibroblasts (HLFs) and EOS, we used the open microfluidic coculture device. This device has two chambers, in which two types of cells can be cocultured in the shared media while being physically separated by a half wall. We found that HLFs in coculture with activated EOS had the highest levels of proinflammatory gene expressions and proinflammatory cytokines. However, the exact mediators responsible for promoting these biological processes are still uncertain. We hypothesize that EOS secrete a cytokine, interleukin-1 alpha (IL-1a), and a protein, transforming growth factor alpha (TGFa), to be consumed by HLFs, triggering proinflammatory responses of HLFs. The goal of this study was to elucidate the roles of IL-1a and TGFa in airway inflammation. HLF-EOS cocultures are seeded in the microfluidic coculture device, then IL-1a, TGFa, and their respective cellular receptors are neutralized using antibodies. Enzyme-linked immunosorbent assays are used to measure the level of EOS-derived neurotoxins after their activation. Then, reverse transcription quantitative-polymerase chain reactions are used to quantify gene expression levels relevant to proinflammatory responses of HLFs, in addition to multiplex immunoassays to analyze the secreted soluble factors from both cell types. We anticipate that HLF-EOS cocultures treated with neutralizing antibodies have lower expression levels of proinflammatory genes than cocultures without antibodies. Findings from this study will help us better understand the key regulators that promote proinflammatory behaviors of HLFs in airway inflammation.

North America is experiencing more frequent and intense wildland fires. Most public health research into exposure of wildland fire smoke is retrospective (backward-looking) examining emergency room visits, hospitalization, and emergency medication usage. In 2021 the Theberge Lab enrolled participants (n=64) in a research opportunity to map and study the inflammatory response of wildland smoke exposure. Participants self-draw liquid whole blood for baseline, smoke events, and 3- and 6-month post season follow-ups. A key advantage of this prospective study is the ability to compare participants’ exposure samples and surveys back to their own pre-exposure baselines. Blood samples are taken by the participants in the comfort of their home using technology developed in our lab called the homeRNA kit. The single use kit allows for a small whole blood sample (100-500 μL) to be withdrawn using the Tasso-SST™, stabilized with RNAlater™ , and mailed to our lab for processing and analysis. While providing samples a series of survey questions are answered. As study coordinator, I assembled sample kits, triggered when to send exposure kits by tracking multiple maps for smoke, shipped and received sample packages, and interfaced with participants. After all samples and surveys are completed, I examined the survey answers in relation to self-reported symptoms, self-reported well-being, general usability, and willingness to use the homeRNA kit during a disaster event. This research intends to provide real-time symptom data from exposure to smoke and to test the feasibility of the homeRNA kit. In the future, the homeRNA kit could be deployed for disaster events, research that requires frequent blood samples, and as a mail-in diagnostic tool between clinics and patients.

The CandyCollect is a lollipop-inspired saliva collection device initially developed to aid in strep throat testing. The device utilizes open, plasma-treated microfluidic channels to collect pathogens in saliva in a noninvasive manner. The device’s isomalt candy coating aids with saliva production, and acts as a built-in timer to ensure adequate oral sampling time. In our previous studies, multiple institutional review board (IRB) approved and IRB exempt studies have been performed to test the functionality of the CandyCollect devices with human subjects. We have also demonstrated the device’s detection of oral bacteria such as Streptococcus pyogenes and Staphylococcus aureus. We noted throughout these studies that the candy’s intended 3 minute dissolving time was longer than expected due to the coating’s large mass. Thus, we are also developing a 1 and 2 minute version to more accurately match the necessary length of sampling times for various pathogens without being unnecessarily uncomfortable for the user. Our goal for this study is to identify the relationship between candy mass, dimensions, and dissolving time so that we can use the candy as a tunable “timer”. We are modifying the depth, diameter, and mass of the device to develop new CandyCollects targeted to take, on average, 1, 2, and 3 minutes to dissolve. These new CandyCollects are more suitable for specific, higher-concentration oral pathogen collections. By reducing the candy’s mass on the devices, we expect the modified devices will decrease the required dissolving time to make the user experience of oral pathogen collection more pleasant. We plan to recruit participants (>18 years) within the university to test these modified devices, and they will provide feedback on their experience and dissolving time. Using these results, we will be able to improve the CandyCollect device to provide a more comfortable sampling experience.

The COVID-19 pandemic demonstrated the importance of screening large numbers of individuals for respiratory pathogens. However, existing sample collection methods, including throat and nasal swabs are unpleasant and invasive, especially for younger patients. The CandyCollect device was designed to alleviate these barriers to efficient sample collection by utilizing specially engineered and surface-treated polystyrene lollipop sticks to capture pathogens from saliva. Prior studies by our team demonstrated the CandyCollect’s effectiveness in collecting commensal bacteria in samples kept stable through ambient temperature shipping, which were then eluted from the CandyCollect and detected and quantified with standard quantitative Polymerase Chain Reaction (qPCR) assays. Here we aim to augment this functionality by testing CandyCollect’s effectiveness with viruses in addition to bacteria. In our Institutional Review Board (IRB) approved clinical study STUDY00013842, we recruited 25 participants from the general public nationwide who were presenting symptoms of any respiratory disease, and sent each a testing kit containing CandyCollect devices in addition to a selection of the current gold standard sampling devices: an oral swab, nasal swab, and a spitting tube. Participants self-collected samples using provided devices and returned kits to our lab for qPCR analysis. Analysis revealed the detection of pathogens Streptococcus pneumoniae, respiratory syncytial virus (RSV A), and rhinovirus (the virus causing the common cold). Of the first five participants with positive signals on at least one of the conventional methods studied, all had concurrent positive CandyCollect signals, suggesting that the rate of successful sample retention of the CandyCollect for these common infection vectors is comparable to that of the currently employed sampling methods. Additional samples continue to be tested with qPCR, and further analysis of these results will be conducted. With this initial effectiveness of pathogen detection, we plan to further streamline the CandyCollect device for younger users, and improve efficiency in clinical settings.

The CandyCollect is a lollipop-inspired device that utilizes microfluidic channels to capture oral bacteria in saliva. The original intent of this device was to successfully collect oral Streptococcus pyogenes (S. pyogenes) from saliva samples and screen for strep throat. Using in vitro experiments, we were able to capture and elute S. pyogenes using the CandyCollect. To explore the functionality of the device, we performed in vitro experiments to capture and elute two different strains of commensal bacteria, Staphylococcus aureus (S. aureus) and Streptococcus mutans (S. mutans). Collection of bacteria using the CandyCollect would allow us to sample from the general population rather than strictly individuals infected with S. pyogenes, allowing the CandyCollect to be used in various applications. The method of bacteria elution we performed was compatible with polymerase chain reaction analysis. We conducted an IRB (institutional review board) approved study to compare the CandyCollect to two other standard saliva collection methods (ESwab™ and SpeciMAX Stabilized Saliva Collection Kit™). Our results showed that the CandyCollect was able to capture both of these commensal bacteria strains when present. We then performed a human subjects study to compare the detection of the CandyCollect to the other two methods. We focused on detection of S. mutans and S. aureus due to their high prevalence in healthy adults. Our results showed that for participants in which a given bacterium (S. mutans or S. aureus) was detected in one or both of the commercially available methods, CandyCollect devices had a 100% concordance with those results. Surveys were sent out to participants to assess the comfort of the sampling methods. The CandyCollect was the preferred method of sampling. Based on the results of this study, we are hoping to one day incorporate the CandyCollect into clinics for strep throat diagnosis to replace more invasive current methods.