Development of a Novel Vaccine to Specifically Treat Autoimmune Diseases

Maya Dahlgren, Fanny Astruc-Diaz, Celine Beamer, Shelby Cole, Joanna Kreitinger, and David Shepherd

Over the last few decades, there has been a significant increase in the amount of people with autoimmune diseases, leading to frequent morbidity and mortality. Immune-mediated diseases, such as lupus, multiple sclerosis, and many others, involve dis-regulation of the immune system, which causes inappropriate attacking of self tissue and organs. Current treatments for these disorders are inadequate and cause many adverse effects, creating a need for a better therapy that can better target and regulate the immune system. Through research we have identified a protein, called the Aryl hydrocarbon Receptor (AhR), that binds PCBs and dioxins which leads to systemic immune suppression. An important population of immune cells that helps to regulate immune responses, called Dendritic cells (DC), express AhR and are highly responsive to AhR activations. From this information, it was hypothesized that if we could deliver activators of AhR to specific immune cells, we could create a drug that is highly specific to suppress unwanted autoimmunity. In order to test this theory, we generated a drug delivery system using liposomal nanoparticles containing AhR activating drugs, autoimmune disease specific peptides, a fluorescent diagnostic marker, and targeting antibodies for mouse DCs. Our results in vitro have demonstrated the utility and selectivity of this system in mouse DCs. Preliminary experiments in vivo have indicated appropriate bio-distribution of the drug-containing nanoparticles when administered to mice. Future experiments are designed to further evaluate the efficacy of this novel immunotherapeutic approach to suppress unwanted autoimmune responses. Ultimately our goal is to develop a highly targeted therapy that can effectively treat autoimmune diseases without compromising the entire immune system.

Twelve species of bats are thought to overwinter in Montana, but surveys have found few individuals hibernating within caves and mines. However, bats are consistently recorded on acoustic detectors year round across the state indicating that they are hibernating in other places, Talus slopes may be a potential hibernacula as there is anecdotal evidence of bats using talus slopes for roosting and foraging. This is a time sensitive issue as disease called White Nose Syndrome, a pathogenic fungus, has been found nearby in Washington. White Nose Syndrome caused severe population declines on the East coast by impacting hibernation behavior. Knowing hibernacula is essential to monitoring and assessing for White Nose Syndrome. Pilot work was conducted during summer 2017 to identify active summer roosts and document species within talus slopes. To continue this work and assess what species are hibernating within these features, we placed ten acoustic recorders on talus slopes to record ultrasonic calls during winter in Northwestern Montana. In warm weather during winter, bats emerge from hibernation to drink, so winter activity may indicate hibernacula in proximity to the detectors. These acoustic data will help us estimate species diversity and relative use at each site. Additionally, we assessed attributes that may influence hibernacula suitability including: aspect, talus depth, talus size, air flow from underneath the slope, and daily temperature and humidity. Out of the ten recorders deployed, only three came back with long-term recordings. These three recorders showed swarming in the fall months and a few calls at each site during the winter months. Both 20khz and 40 khz bats were recorded showing some species diversity and they were predominantly recorded at duck, but also during the night. While there were not enough recordings to complete a microclimate analysis, this data shows that bats are hibernating in close proximity to the detector. This data helps us to establish a baseline understanding of where our bats are hibernating before White Nose Syndrome comes, so we can better assess and mitigate the fungus.

The bacterium Borrelia burgdorferi is the causative agent of Lyme disease, the most common arthropod-borne disease in the Northern Hemisphere. This ailment affects 400,000 people annually, and can cause symptoms such as fever, joint stiffness, fatigue, carditis, and neurological issues. One of the most fundamental processes in any cell, including B. burgdorferi, is the transcription of DNA into an RNA messenger, which is later translated into proteins. Transcription is carried out by a multisubunit molecular complex called RNA polymerase, which is recruited onto the DNA strand and synthesizes RNA by reading the DNA. RpoD, also known as σ⁷⁰, is the subunit of RNA polymerase in bacteria that recognizes the correct binding site on DNA. In all bacteria studied to date, the size of RpoD is 70 kilodaltons (kDa). Although the rpoD gene in B. burgdorferi appears to encode a 70-kDa subunit, two lines of evidence suggest that only a 50-kDa protein is produced in these bacteria. In order to biochemically dissect the function of this truncated RpoD in B. burgdorferi, I have overexpressed and purified it to near homogeneity using recombinant DNA methodologies. The putative RpoD protein was then assayed for transcriptional activity using an in vitro transcription assay. Future directions for this project would include obtaining a crystal structure of the truncated RpoD from B. burgdorferi and analyzing its regulation.

A bacteriophage integrase regulates virulence factor production in Pseudomonas aeruginosa

Pseudomonas aeruginosa is a bacterial pathogen that causes hospital-acquired infections and is difficult to kill with antibiotics. This is especially true when P. aeruginosa grows as a biofilm—a complex community of bacteria encased in a protective extracellular matrix. P. aeruginosa biofilms produce large amounts of filamentous Pf bacteriophage, which are viruses that infect P. aeruginosa. Pf bacteriophage are known to increase the virulence of P. aeruginosa; however, the underlying mechanisms are unknown. Our preliminary results suggest that when the Pf bacteriophage integrase gene, intP, is overexpressed, production of the virulence factor pyocyanin is enhanced. When intP is deleted, pyocyanin production is repressed in P. aeruginosa. Like other bacteriophage integrases, intP inserts bacteriophage DNA into the bacterial chromosome. We hypothesize that intP integrates bacteriophage DNA into bacterial genes that regulate pyocyanin production. To test this hypothesis, we will use genetic approaches to disable the integrase activity of intP followed by biochemical assays to measure pyocyanin production. Understanding how Pf bacteriophage manipulate the virulence of P. aeruginosa may result in new ways to treat or prevent infections caused by this ubiquitous bacterial pathogen.

An important component of understanding the dynamics and stability of communities is to elucidate how interspecific species interactions respond to various ecological disturbances. We address this issue by removing a dominant floral resource, Helianthella quinquenervis (Asteraceae) from an alpine meadow and observing the effects on plant-pollinator interactions. Specifically, we examined how this removal effected pollinator visitation, pollinator community structure, and co-flowering plant reproduction. We addressed three primary questions: (1) How do pollinator visitation rates vary between unmanipulated floral communities and those where the dominant floral resource is removed? (2) How does pollinator diversity and community composition differ between natural and manipulated communities? and 3) How does variation in pollinator visitation influence the reproduction of the two other co-dominant, flowering plant species? To study these components, we conducted paired plant-pollinator observations in control and removal plots for 30 hours per week for 5 weeks. We then analyzed differences in pollinator visitation rates between control and removal plots using paired t-tests and linear mixed effects models. At the community level, we found that overall pollinator visitation rates increased in response to resource availability. At the species level, the removal increased visitation to one common co-flowering plant species, and decreased visitation to another. Our findings illustrate how the removal of a dominant resource can alter the dynamics of plant-pollinator interactions and provide insight into how ecological communities respond to disturbance.

The Northern Goshawk is listed as a management indicator species for the Minidoka Ranger District of the Sawtooth National Forest. This distinction has enhanced research interest on goshawk population health in the region. For raptors, annual adult turnover is considered a crucial metric of population health; providing insights into mortality, fidelity, and population disturbances. Over the past 25 years of studying goshawks, the Intermountain Bird Observatory (IBO) has observed abnormally high female turnover as compared to other places the species has been studied. Their estimations are based on banding and resighting birds, and may be biased high due to undetected marked birds and unknown age of birds when banded. To increase accuracy of IBO’s turnover data, we conducted parentage analyses using blood samples collected from goshawks in 2012-2016. We analyzed 32 samples from nine nest territories by examining shared alleles between adults and nestlings. With this analysis, we identified previously unknown turnover and fidelity events, increased known ages of banded birds, and quantified and removed bias from IBO’s turnover estimations. Our work indicated that band-resight alone may be insufficient to produce accurate turnover estimates, and the inclusion of genetic analyses may mitigate inaccuracies. In addition, our results fundamentally altered IBO’s understanding of goshawk population dynamics within the forest.

Imperfect detection is ubiquitous among wildlife research and can affect research conclusions and management. Accordingly, detection probability is often included in observation-based models. Here, we leveraged long-term research of gray wolves (Canis lupus) in northern Yellowstone National Park to evaluate how the probability of sighting radio-collared wolf packs from ground-based locations was affected by the characteristics of each spatial location (i.e., distance from the road and viewability [a combination of landscape “openness” and whether visible from a viewshed created from the road and nearby observation points]). To do so, we used 2,681 unique, daily observations of 17 wolf packs collected during 44 unique 30-day winter monitoring periods from 1995 – 2017 and used matched-case control logistic regression with a 1:1 sampling design between observed and random locations. We found that the probability of wolf sightings declined as wolves were farther from the road and increased when wolves were in open, viewable areas. We then evaluated whether these conclusions were affected by wolf group size or whether wolves were feeding at a carcass and found that the probability of sightings only clearly decreased when smaller groups of wolves were farther from the road. Ultimately, we used our results to build spatial predictions for seeing radio-collared wolves in northern Yellowstone National Park. These predictions are useful to managers by identifying “hot-spots” of wolf observations, and can also be incorporated into research related to wolf ecology and predator-prey dynamics that relies on ground-based observations of wolves.

Sexual selection theory states that male ornamentation may evolve if it helps males obtain more matings by means of male-male competition or female preference. Dominant males can monopolize limited resources to attract mates, increasing variance in male reproductive success and strengthening the effects sexual selection. While there have been several studies examining the function of ornaments in intraspecific contests, less is understood about the role of ornamentation in interspecific competition. In a population of mountain bluebirds near Ronan, MT, tree swallows arrive after mountain bluebirds are nesting, and compete directly for access to nest-boxes. A successful tree swallow intrusion often results in total brood mortality for bluebirds, so the ability of a male mountain bluebird to defend a territory is directly linked to his and his mate’s fitness. Male mountain bluebirds have structural UV-blue plumage and there is evidence that more saturated coloration is associated with increased success in defending territories. We conducted 30 minute observations at active bluebird nests and recorded the parental behavior of bluebirds such as nest attendance and feeding rate. Observations also recorded the length of tree swallow intrusions, number of intruders, and bluebird behavioral response. Through this research, we aim to explore the relationship of male mountain bluebird coloration to his and his mate’s behavior and the characteristics of tree swallow intrusions. Our findings not only provide insight into the effect of ornamentation on interspecific competition, but also serve to support hypotheses regarding status-signal honesty.

I will investigate the club and suffrage work of Waltrene Willis, a resident of Thomson Falls, Glasgow and Garfield Montana between 1900 and 1920, who worked as the recording secretary for the Montana Equal Suffrage State Central Committee. I conducted research in federal census records, publications of the Glasgow Courier and The Suffrage Daily News, and the contemporary publication of the Montana Woman. My research on Waltrene Willis illustrates the range of activities that were conducted by Progressive-era women in Montana. Involved in everything from the Sunflower Art Club to Glasgow’s Political Equality League, Willis’s career serves as an example of the work women did to achieve suffrage. Understanding the activities of the women devoted to the cause of suffrage in Montana is critical in furthering our knowledge of Montana women’s history.

This research examines the life and work of Margaret Jones Sounders, a Montana woman active during the suffrage movement in the state. Souders was identified as a key suffragist in the state of Montana in the History of Woman Suffrage book, originally published in 1881. The research for this project was conducted through the use of genealogy sites Ancestry and Heritage Quest, historical Montana newspapers, digitized materials included in the Montana Memory Project, and documents from the Montana Historical Society. Little if any research has been conducted about the majority of individuals included in the History of Woman Suffrage. We learn more about Souders’s life and suffrage work by tracing her movement across the state of Montana and her interactions with various clubs and organizations. Through this project we can understand that Souders played a role as a figure in the Montana Federated Women’s Clubs and encouraged more women to get involved Her life reveals previously hidden aspects of the suffrage movement at the state level.

Calyn Hitchcock

Margaret Jane Rozsa, A Montana Suffragist

I will explore the life and activism of Margaret Jane Rozsa, a resident of Butte, Montana. Rozsa was not only an advocate of woman suffrage but also a strong proponent of accessible education and social services. She also was involved in the Progressive-era anti-prostitution movement. To learn about her life, I conducted research in federal census records, Montana newspapers, the California Death Index, and U.S. City Directories. My research on Margaret Jane Rozsa highlights many of the issues women in America cared about during the Progressive Era. By focusing on a Montana suffragist, it also offers a glimpse behind the scenes of the national suffrage movement.