Mike Czapka

Home School: Loyola University-Chicago
Loyola Mentor: Dr. Wiethoff/Kathleen McGuire

Kelsey Flood

Home School: University of Notre Dame
Loyola Mentor: Dr. Caroline LePoole

This summer I had the opportunity to work in the lab of Dr. Caroline Le Poole who studies the role of HSP70 in vitiligo development. Vitiligo is an autoimmune disorder characterized by depigmentation as a result of CTL killing of skin melanocytes. My project involved quantifying HSP70 secretion in healthy and vitiligo melanocytes and determining its cellular localization alongside the graduate student Jeff Mosenson. Using techniques such as ELISA I will measure levels of HSP70 in healthy and vitiligo melanocytes in response to bleaching phenols. Moreover, I used four-color confocal microscopy to observe colocalization of HSP70 within melanosomes by staining for TRP-1, MART1 and HMB45. To further investigate whether HSP70 is present within melanosomes, I loaded melanocyte cell gradients on a SDS-PAGE gel and stained for HSP70 and various melanosomal markers via Western Blot. Finally, I am detecting the presence of HSP70 in vitiligo skin sections via immunohistochemistry.

Therese Geraghty

Home School: Michigan State University
Loyola Mentor: Dr. Iwashima/Dr. Love

I worked in Dr. Love’s lab this summer looking at gene expression in mice lung transplant models. We’ve found that if levels of IL-17, a pro-inflammatory cytokine in Th17 responses that causes airway remodeling, are treated with OmentalStromal Cells (OSC) that inflammation is reduced. This was found in mice lung tissue and so my project was to study the gene expression in BAL lavage fluid. The results we were getting showed no pattern and we concluded that what we were seeing in the lung tissue is probably not being released in the lung airway and that’s why we’re not seeing the same thing in the lavage fluid. So I moved on to a new project involving the COL5A1 gene. Our lab has previously found that COL5A1 is implicated in allograft rejection and dysfunction in lung transplantation. My part in this project was to analyze donor recipient matches through Restriction Fragment Length Polymorphisms (RFLPs). I used a similar protocol involving isolating genomic DNA, running PCR’s for each of the donor and recipients, restricting fragments through enzyme digestion and separating those fragments on an agarose gel using a 100bp ladder. Once the data I collected is analyzed, our lab will be able to complete the statistical analysis and determine whether protein pattern expression differs in response to host tissue injury.

Leandra Knapp

Home School: University of Notre Dame
Loyola Mentor: Dr. Knight/ Greg Robbins

This summer I worked with PhD candidate Pehga Johnston in Dr. Knight’s lab on studying Clostridium difficile, which is a largely antibiotic-resistant colon bacterium. Pehga’s previous experimentation has shown that mice re-challenged with C. difficile, after recovering from a previous episode, are resistant to the harmful effects of the toxins released by C. difficile, suggesting there is a protective immune response. My project was centered around the following question: How does anti-toxin antibody mediate cell protection (against toxin)? It has been commonly thought that anti-toxin antibody merely neutralizes Toxin, but recent research has shown that the FcR may be required. The first step to answering my question was to purify Toxin using protein precipitation and anion exchange chromatography, and then use MTT assays to confirm enzymatic activity. In my last couple of weeks in the lab, I will be testing the “neutralizing” hypothesis by using MTT assays with anti-toxin antibody added to the assay, and if this is shown to not be effective, further experimentation will be done in vivo to determine whether a complement receptor or FcR is required.

Colin Linke

Home School: Loyola University-Chicago
Loyola Mentor: Dr. Karen Visick

Madeline Lyons

Home School: Duke University
Loyola Mentor: Dr. Lanning

This summer, I had the pleasure of working under Dr. Dennis Lanning to study the pathway of development of the neonatal rabbit antibody repertoire. Based on previous experiments, we knew that different intestinal microbiota interacted with B cells in the follicles of the rabbit appendix to either stimulate or not stimulate proliferation of B cells, leading to antibody diversification. Our goal this summer was to develop a method by which we could visualize the uptake of bacteria into the rabbit appendix to better understand how the B cells and the bacteria interact and how different bacteria can differentially cause B cell proliferation. Through in situ hybridization and immunohistochemistry, we aimed to visualize the interaction between these B cells and the intestinal commensals of interest. With this method of visualization, we will be able to study important aspects of the B cell proliferation pathway.

Nathan Manthey

Home School: Bethel University
Loyola Mentor: Dr. Iwashima/Dr. Love

This summer I worked in the Thoracic CV Surgery lab, where my project focused on omentum. My project involved obtaining omental cells (OCs) from mice and treating them with various media to differentiate them into lung epithelium-like cells. For my project, I dealt with a sorted magnetic bead culture. More or less, the sorting protocol for this culture helped me separate the three different subpopulations (CD45+, CD45-/CD34+, and CD45-/CD34-) of omentum. The main focus for my study was to determine the subset of omentum cells that differentiate into lung epithelium-like cells. To demonstrate that a subset of cells differentiates, I took weekly cell counts for 32 days. My hypothesis was that if a single subset divided by CD45/CD34 expression is responsible for lung epithelium cell differentiation, then the single isolated OC subset is sufficient to develop into cells expressing CCSP and Surfactant Protein A & C. Overall, the cell counts revealed that the CD45-/CD34+ subpopulation does differentiate into lung epithelium-like cells. For future work on this project would be to obtain western blot results that show the expression of various proteins typical of lung, such as CCSP and Surfactant Proteins A & C. These results would confirm that the CD45-/CD34+ subpopulation was, in fact, differentiating into lung epithelium-like cells.

Home School: Waubonsie Valley High School
Loyola Mentor: Dr. Lanning

I worked with Dr. Dennis Lanning this summer to research the transcription factor Foxn1 in mice. Foxn1 maturates TECs and promotes T cell development, yet the levels of Foxn1 deplete with age, causing effects such as a lower naïve T-cell output, a lower number of T cell progenitors, and a lower number of T cell precursors. It is known that the lack of Foxn1 in mice causes the loss of hair follicles (a nude mouse) and athymia. The mice Dr. Lanning, Dr. Phong Le, and I have worked with over express Foxn1 and thereby do not manifest the age related symptoms of regular mice. My experiment is centered around the hypothesis that Foxn1 is produced in the same cells in over expressed mice and wild type mice. To do this, I first created a riboprobe specific to the Foxn1 transcription factor; now, I am performing in situ hybridization on MsFoxn1 OE thymus and skin tissues to fluoresce localized Foxn1.

Ashley Schmitz

Home School: Marian University
Loyola Mentor: Dr. Zeleznik-Le

Throughout this summer I have been studying MLL leukemia with the assistance of Alyson Lokken, a PhD student in Dr. Zeleznik-Le’s oncology lab. We have been focusing on identifying areas of methylation on fibroblast cells within a time range of 0-13 days, and methylation, when present, plays a crucial in the prevention of the binding of activating proteins. Within this project I was able to gain experience performing many experimental techniques, such as splitting and maintaining cell colonies, using vectors to infect cells with specific regions of RNA through both chemical and electroporation, using PCR to amplify certain regions of DNA and RNA, and other invaluable skills and knowledge that I will certainly use during my graduate education and after. I have grown immensely as an aspiring scientist and am so grateful for this experience and everyone I worked with.

Stephen Small

Home School: Butler University
Loyola Mentor: Dr. Knight/Dr. Jones

I had the opportunity to work with Dr. Sara Jones in the Knight Lab studying why certain species of Bacillus bacteria have probiotic effects in mice. Citrobacter rodentium causes an enteric disease in mice that is similar to Enteropathogenic and Enterohemorrhagic E. coli infections in humans.  It has been found that Bacillus subtilis and B. licheniformis spores protect mice from C. rodentium infection whereas B. formis spores do not. Also, it has been shown that these bacteria must possess flagella and exopolysaccharide in order to be protective. To better understand the differing degrees of protection between these species, my research focused on studying how fast these Bacillus species are able to produce protective flagella and exopolysaccharide.  This was done by measuring and comparing the rate of biofilm formation and spore germination—in vitro and in vivo—for each species.  While preliminary, our results suggest that B. subtilis and B. licheniformis spores produce these protective factors more rapidly by germinating faster than B. formis spores.

Home School: Cedarville University
Loyola Mentor: Dr. Thomas Gallagher

Virus infections are confronted by mobilizing a defensive arsenal that is orchestrated by the interferons (IFNs).  There are hundreds of IFN-stimulated genes whose products restrict virus infections at virtually every conceivable stage of the infection cycle.  Amongst the broadly – acting IFN-induced restriction factors are the IFN-Induced Trans Membrane proteins (IFITMs).   These IFITMs are small membrane proteins that restrict the entry of a wide variety of enveloped viruses including alpha, flavi, rhabdo, filo, paramyxo, orthomyxo, retro and coronaviruses. This summer, my mentor Dr. Thomas Gallagher and I had the opportunity to further research the role that IFITM3 plays in the virus entry restriction. My research asked the specific question, “Do IFITMs restrict murine and human coronaviruses at the level of membrane fusion?” To address this question, I used array of techniques to measure virus-cell and cell-cell fusion at the plasma membrane. By constructing viruses and cells with or without IFITMs, and by measuring the effects of the IFITMs on membrane fusion events, I was able to further reveal how these small membrane proteins restrict coronavirus entry.  Further advancing the scientific community’s understanding of virus restriction, this research also translates to clinical medicine where it provides valuable information to biomedical researchers interested in anti-viral drug development.