Immunology and Infectious Disease Commons^™

Development Of Wiskott-Aldrich Syndrome Knock Out Protocol For Drug Substance Assay Development, Julia C. Hanna

Master's Theses

Wiskott-Aldrich Syndrome (WAS) is a rare X-linked primary immunodeficiency affecting approximately 1 in 100,000 live XY births in North America and is caused by a mutation to the WAS gene which is expressed across hematopoietic lineages. The WAS protein (WASp) plays a role in regulating actin polymerization. On a cellular level, there are a variety of effects of a lack of WASp or expression of a dysfunctional WASp protein for patients including issues with migration, adhesion, chemotactic response, phagocytosis, activation, and proliferation across different cell types in addition to reduced platelet size and output. This can lead to several systematic …

Sensitivity Of A Lamp Assay For Detection Of The Dinoflagellate Amyloodinium Ocellatum In Simulated Field Conditions And Freeze Tolerance Of The Parasite, Robert Gonzales

Master's Theses

The obligate parasitic dinoflagellate Amyloodinium ocellatum causes amyloodiniosis in warm water marine fishes. The prolific parasite, which has a direct, three-stage life cycle, is highly infectious and can cause heavy losses in aquaculture. Prevention, biosecurity, and early detection are vital for control. In this work, microscopy and a loop-mediated isothermal amplification (LAMP) assay were compared for early diagnosis of A. ocellatum in cultured stocks, and the freeze tolerance of tomonts was assessed to determine if frozen wild fish used as fish food can serve as a potential vector for the parasite.

The lowest dinospore concentration that could be detected by …

The Role Of Ly49 Receptors In Maintaining Healthy Murine Pregnancy, Michelle Belmont

Master's Theses

Natural Killer (NK) cells are abundant in pregnancy and contribute to the health of the fetus. Uterine NK (uNK) cells provide less cytotoxic function than their conventional NK (cNK) cell counterparts. Various combinations of Killer Immunoglobulin Receptor (KIR)- Human Leukocyte Antigen (HLA) on maternal NK and fetally derived cells, respectively, can have different effects on the health of pregnancy in humans. Murine NK cells contain Ly49 receptors, an equivalent to KIRs. Therefore, Ly49 receptor interactions with fetally-derived MHC-I molecules can be a valuable model for studying the role of NK cells in pregnancy. The process of spiral artery remodeling can …

Phenotypic Analysis Of Murine Gut Associated Lymphocytes, Madison Flood

Master's Theses

γδ T cells are important in the role of immune homeostasis, especially in relation to the gut. It has been shown that there is a higher proportion of γδ T cells in the intestines than in circulation. With this knowledge, our lab focused on understanding the mechanisms of these cells, specifically regarding immune tolerance. In our preliminary research, we identified a subset of γδ T cells that express Foxp3, a transcription factor essential for the functioning of regulatory T cells, in the neonatal (5 day old) intestine. We hypothesized that these Foxp3+ γδ T cells have immunosuppressive functions upon exposure …

Plasmodium Tlap-2 Is A Microtubule-Associated Protein With Microtubule Stabilization Properties, Grifin Berge

Master's Theses

Microtubules facilitate major cellular and morphological changes in eukaryotic cells, including the protozoan parasite Plasmodium. Microtubule-associated proteins (MAPs), only very few of which have been characterized in the malaria parasite, regulate the highly dynamic microtubules. Thioredoxin-like associated protein 2 (TLAP2) is a MAP unique to the phylum apicomplexa, which in addition to Plasmodium includes the important human pathogen Toxoplasma. Here we report on the cloning and microtubule-binding activity of TLAP2 of the rodent malaria parasite Plasmodium berghei (PbTLAP2). PbTLAP2 is coded by a single exon gene and consists of 598 amino acids. The protein contains a putative C-terminal microtubule-binding domain. …

Effects Of Infection Of The Protist Parasite, Dermomycoides Sp., In Dusky Gopher Frog Tadpoles, Jaime Smith

Master's Theses

Infections of the protist parasite, Dermomycoides sp. are thought to have caused several years of low recruitment in the dusky gopher frog (Rana sevosa) populations. I evaluated the effects of density of the infective zoospores, host developmental stage, and tadpoles' ability to acquire resistance to Dermomycoides sp. on dusky gopher frog tadpoles. Tadpoles were exposed to zoospore densities of 0, 250, 500, and 750 zoospores/µL at Gosner stage 25, and we found no significant differences among treatments in tadpole mortality. In evaluating susceptibility by development stage, I exposed R. sevosa to 50 zoospores/µL as eggs, embryos, hatchlings, and …

Characterizing Erad And Antioxidant Response In Ixodes Scapularis Ise6 Cells During Borrelia Infection, Latoyia Downs

Master's Theses

Ticks are hematophagous ectoparasites that transmit a multitude of diseases to humans. Borrelia burgdorferi (BB) and Borrelia miyamotoi (BM) are both tick-borne pathogens that cause disease in humans and are transmitted by the black-legged tick (Ixodes scapularis). A byproduct of blood digestion generates reactive oxygen species that are toxic and cause oxidative stress which promotes cellular damage and dysfunction. The endoplasmic reticulum (ER) is especially affected by oxidative stress, resulting in a buildup of improperly folded proteins in the ER lumen called ER stress. To prevent cellular damage, the tick utilizes an antioxidant system to neutralize ROS and mechanisms …

Development Of An Attenuated Zika Virus By Editing The 5’ Untranslated Region, E. Ashley Thompson

Master's Theses

Zika Virus (ZIKV) is a mosquito-transmitted flavivirus that usually causes no symptoms to mild febrile in humans, and it has been regarded as an insignificant pathogen to public health. However, recent outbreaks of ZIKV infection have revealed that ZIKV can cause severe neurological effects in adults, such as Guillain-Barre Syndrome (GBS), and in infants whose mothers acquired the virus during pregnancy, causing Congenital Zika Syndrome (CZS). Currently, no approved vaccine is available and there is a critical need to develop an effective and safe vaccine. While most vaccine developmental strategies target the viral prM-E protein of ZIKV, we aimed to …

The Characterization Of Inflammatory Extracellular Vesicles, Barak Balva

Master's Theses

One of the driving factors behind innate immunity is the early response of inflammation. While beneficial in controlling infections, when uncontrolled it can lead to a plethora of disorders. Thus, it is essential to understand the driving factors behind inflammation. a cell's ability to recycle, discard, or transfer proteins is integral for survival and intercellular communication. It has been appreciated that inflammatory components, such as caspase-1, play a huge role in driving protein secretion. Thus, inflammation also plays a role in the distribution of proteins released from cells. These proteins are released via non-conventional secretory pathways, and our lab is …

Transforming Growth Factor Beta Suppression Of Cd8+ T Cell Proliferation, Shena Geisinger

Master's Theses

Transforming Growth Factor Beta (TGF-beta) is highly suppressive to both CD4+ and CD8+ T cell proliferation and function. in tumor microenvironments, TGF-beta has been described as immune suppressive, particularly to CD8+ T cells, however, the molecular mechanism behind how TGF-beta signaling controls T cell growth is not fully understood. Here, we report that TGF-beta inhibits CD8+ T cell proliferation and reduces expression of the CARMA1/BCL10/MALT1 (CBM) signalosome complex in activated CD8+ T cells. the CBM signalosome is an essential scaffold that forms after T cell receptor (TCR) stimulation, leading to the activation of NF-kB and AP-1. This observed reduction in …

Characterization Of Caspase 1 Substrate Specificity Utilizing Luminescent Reporter Constructs, Nicholas Rohlfes

Master's Theses

Inflammation is the bodies first response to defend itself against foreign pathogens and damage. Cellular sensors are constantly monitoring the body and checking for homeostatic disruption. Microbial pathogens and tissue damage are sensed by pattern recognition receptors (PRRs), which sense pathogen associated molecular patterns and damage associated molecular patterns, termed PAMPs and DAMPs respectively. Within the family of PRRs are sensors that can induce the formation of the multi-protein complex termed the inflammasome following activation. the inflammasome complex is a molecular platform for which caspase 1, a cysteine protease, is incorporated and is responsible for the maturation of pro-inflammatory cytokines …

Enabling The Clinical Detection Of Human Papilloma Virus (Hpv) To Enhance Effective Diagnosis And Research, Jennifer Sackey

Master's Theses

Human Papilloma Virus (HPV) is one of the most common sexually transmitted infections. There are over 150 different genotypes of HPV. Some cause benign warts, while others cause several types of cancer. Subunit vaccines comprised of the capsid protein from an increasing number of HPV genotypes have been developed; however, the vaccines are prophylactic and are documented to protect against only 9 of the HPV genotypes. Importantly, there are also significant limitations in the clinical detection of HPV. Current clinical tests can only specifically identify HPV16 and HPV18. One of the test also detects 12 of the other high-oncogenic risk …

Induction Of Zika And Dengue Virus-Specific Cd8+ T Cells By Means Of Dna Vaccine, Natalie Jachym

Master's Theses

Zika virus (ZIKV) and Dengue virus (DENV) are flaviviruses that circulate in the same endemic regions and can co-infect the same individual. While primary infection by DENV leads to life-long immunity to the infecting serotype, it does not provide immunity to other serotypes. Moreover, ZIKV and DENV antibodies can be cross-reactive and potentially enhance infection of the other virus through a process known as antibody-dependent enhancement (ADE). In this mechanism, antibodies weakly bind the infecting virus and direct virions to immunoglobulin Fc receptors on macrophages. This promotes viral uptake and infection rather than neutralize the virus. Due to this mechanism, …

Development Of A Prolyl Endopeptidase Expression System In Lactobacillus Reuteri To Reduce The Clinical Manifestation Of Celiac Disease, Kara Lynn Jew

Master's Theses

Celiac Disease (CD) is an autoimmune disorder that emerges due to the ingestion of gluten, a protein found in a variety of common grains such as wheat, rye, and barley. Approximately 1 in 100 individuals in the US suffer from CD, making it the most commonly diagnosed gastrointestinal disorder (Ciclitira et. al., 2005). These proline-rich gluten peptides are resistant to proteolysis and accumulate in the duodenum of the small intestine. Once in the duodenum, these peptides illicit an autoimmune response resulting in villous atrophy. Current treatment for CD requires a rigorous adherence to a gluten-free diet. Nevertheless, gluten-containing grains are …

Attenuated Responses To Inflammatory Cytokines In Mouse Embryonic Stem Cells: Biological Implications And The Molecular Basis, Bohan Chen

Master's Theses

Embryonic stem cells (ESCs) have attracted intense interest due to their great potential for regenerative medicine. However, their immune property is an overlooked but a significant issue that needs to be thoroughly investigated not only to resolve the concern for therapeutic applications but also for further understanding the early stage of organismal development. Recent studies demonstrated that ESCs are deficient in innate immune responses to viral/bacterial infections and inflammatory cytokines. Inflammatory conditions generally inhibit cell proliferation, which could be detrimental to ESCs, since cell proliferation is their dedicated task during early embryogenesis. Thus, I hypothesize that the attenuated innate immunity …

Induction Of Zika Virus-Specific Cd8+ T Cells By Means Of Dna Vaccine, Frank Gambino Jr.

Master's Theses

Zika virus (ZIKV) is a member of the virus family flaviviridae and is transmitted via Aedes aegypti mosquitos. Monoclonal antibodies against dengue virus have been found to cross-react with ZIKV but show no ability to neutralize it. Further studies have shown that Stat2-/- mice given plasma from dengue positive donors exhibit a heightened disease phenotype when challenged with ZIKV. Antibody dependent enhancement is theorized to cause such effect. The goal of this study was to design a plasmid which, when encoded, creates an antigen that will be used to stimulate exclusively CD8+ T cells. The gene sequence of NS3 was …

Cell Intrinsic Characteristics Of Cord Blood Naive Cd4 T Cells, Taylor Keller

Master's Theses

The neonatal immune system is functionally distinct from the adult immune system. Neonatal immune responses are less reactive than their adult counterparts, and as such, have an increased susceptibility to infection, resulting in increased duration and severity of disease. There is evidence that T-cells are partially responsible for the observed immunological differences between neonates and adults. Murine studies on neonatal immunity suggest that Th2 like responses predominate the neonatal immune system, an observation not found in the adult immune system. However, studies on human cord blood revealed that human neonates do not exhibit elevated Th2 like cytokines (with the exception …

Lidocaine Attenuates An Induced Inflammatory Response, Stephanie Zack

Master's Theses

Inflammation is common to the pathogenesis of a variety of diseases. Therapeutics which can regulate and reduce damaging inflammation are therefore valuable in the treatment of inflammation and inflammatory diseases.

One commonly used analgesic, recently identified as limiting inflammation, is lidocaine. Additionally, alternative, less toxic therapeutics like the snake venom peptides, Zep 3 and Zep 4, are thought to have anti-inflammatory effects. Yet a well-defined mechanism or cellular target for lidocaine or Zep peptides' anti-inflammatory effects has not been proposed.

We aimed to determine whether lidocaine and Zep peptides reduce the release of the proinflammatory cytokine, IL-1β, from activated human …

The Characterization Of Rna Content And Biogenesis Pathways Of Extracellular Vesicles That Have Been Implicated In The Pathogenesis Of Hiv-1 Associated Neurocognitive Disorders, Virginia Elizabeth Zwikelmaier

Master's Theses

Patients infected with HIV-1 adhering to Anti-Retroviral Therapy (ART) still suffer from a variety of neurocognitive, behavioral, and motor dysfunctions, collectively referred to has HIV-1 Associated Neurocognitive Disorders (HAND). Current research suggests the manifestations of HAND might in part be due to the dissemination of toxic HIV-1 components throughout the brain via extracellular vesicles (EVs). Our goal is to characterize these pathogenic EVs to begin to gain a better understanding of this EV-mediated pathology.

We took the first steps in developing a technique to assess the microRNA content of individual EVs but it needs to be heavily optimized to eliminate …

The Novel Characterization Of Extracellular Vesicles Containing Proteins Which Have Been Implicated In The Pathogenesis Of Hiv Associated Neurocognitive Disorders, Grace Ispas

Master's Theses

Although Anti-Retroviral Therapy (ART) is practiced, HIV-1 positive individuals still experience HIV-Associated Neurocognitive Disorders (HAND), collectively described as the presentation of neurocognitive, behavioral and motor dysfunctions that decrease the quality of life, while increasing the mortality in ART suppressed HIV-1 positive patients.

Current literature suggests that extracellular vesicles are involved in the pathogenesis of HAND as they are believed to be transferring HIV-1 proteins to nearby neuronal cells. Although most studies assume homogeneity among populations, characterizing cellular proteins or RNA levels in bulk, we hypothesize that distinct populations of extracellular vesicles are released and that environmental conditions, including viral infections, …

Dexamethasone Treatment Effects On H3k27me3 Chromatin Organization Is Related To Nk Cell Immune Dysregulation, Michael Sarafeno Misale

Master's Theses

It is well-established that psychological stress reduces natural killer (NK) cell immune function. This reduction is mediated by stress-induced release of glucocorticoids (GC), which can suppress immune function. Associated with suppression of a particular immune function are GC induced histone-epigenetic marks. Histone-epigenetic marks are responsible for the organization and compartmentalization of genomes into transcriptionally active euchromatin domains that are localized to the interior of the nucleus. Transcriptionally silent heterochromatic domains are enriched with methylated epigenetic marks and are localized to the nuclear periphery. The purpose of this investigation was to assess the influence of GC on H3K27me3 chromatin organization by …

Multifunctional Regulatory T Cells From Human Umbilical Cord Blood And The Role Of Tumor Necrosis Factor In Immune Homeostasis, Alexander Nelson

Master's Theses

Regulatory T cells (Tregs) are required to suppress inflammation and prevent autoimmunity. During fetal development Tregs are crucial to maintain tolerance between mother and child. After birth, neonates require tolerance to avoid harmful immune responses to foreign antigens in food and allow colonization with commensal microbes. We demonstrate a propensity for T cells in human umbilical cord blood to differentiate into Tregs in response to antigen receptor stimulation ex vivo. Cord blood-derived Tregs potently suppress T cell proliferation, but also produce pro-inflammatory cytokines known to activate innate immune responses. These results suggest that antigen exposure during early life results in …

Characterization Of Il-22-Producing Cells In The Human Thymus And The Mechanism By Which Il-22 Expression Is Modulated By Thymic Epithelial Cells., Nadine N. Morgan

Master's Theses

Thymic epithelial cells (TECs) are indispensable for T cell development and maturation. Therefore, damages to the thymic epithelial cells are detrimental to thymic function and immune response. In young healthy individuals, TECs have a high regenerative potential and are capable of renewal from serious damage; however, the molecular mechanism of this recovery is unclear. A recent study has shown that IL-22-producing ILCs are present in the mouse thymus and can regenerate thymic epithelial cells following radiation induced injuries. However, it is unknown whether IL-22-producing cells are also found in the human thymus. Thus, the goal of this study is to …

A Study Of Immune-Regulation Of Microbiota Supernatant On Macrophage Function, An N. Nguyen

Master's Theses

The diverse human microbial ecosystem colonizes many different body sites. The gastrointestinal (GI) tract, especially, is a major region that that harbors a complex microbial community composed of trillions of commensal bacteria. Interactions between the commensal bacteria and the host immune system begin at birth and continue through the human lifespan. In the gut, commensal bacteria help the host to ensure the maintenance of intestinal homeostasis as well as the mucosal barrier structure through host stimulation of mucus and antimicrobial molecules. The secretion of these host factors functions to inhibit pathogenic invasion and maintain the population of commensal bacteria. The …

Defining The Role Of A Putative Peptidase In Staphylococcus Aureus Quorum Sensing And Pathogenesis, Chelsea Rose White

Master's Theses

The pathogenic bacterium Staphylococcus aureus encounters a variety of immune cells that restrict bacterial growth and dissemination during infection. However, S. aureus counters these immune cell functions by producing myriad virulence factors that facilitate evasion of host defenses. Therefore, it is important to understand S. aureus-immune cell interactions and the virulence factors that perturb normal immune function to promote pathogenesis. In a genetic screen designed to identify S. aureus secreted factors that modulate macrophage activity our lab identified a transposon insertion mutant in the gene SAUSA300_1984, encoding a putative membrane spanning peptidase. To investigate the role of 1984 in pathogenesis, …

Detection Of Hcmv Viral Il-10 (Vil-10) In Healthy Blood Donors, Vivian P. Young

Master's Theses

Human cytomegalovirus (HCMV) is widespread in the general population and can establish lifelong latency with expression of a limited subset of viral genes. The UL111A gene is expressed during both lytic and latent infection, and at least two protein isoforms are produced. During lytic infection, the full length transcript yields cmvIL-10, a potent immunosuppressive viral ortholog of human IL-10 (hIL-10). Alternative splicing of the UL111A transcript yields a truncated protein, LAcmvIL-10, which is expressed during both lytic and latent infection but with a limited range of immunosuppressive functions. The two viral cytokines, collectively termed viral IL-10 (vIL-10), are identical in …

Role Of Heat Shock Proteins In Maintenance Of The Gut Barrier Following Burn Injury, Abigail Rhea Cannon

Master's Theses

Introduction: Burn injury remains a prominent clinical problem. Patients suffering from burns often succumb to secondary infectious complications leading to sepsis and widespread tissue injury ending in Multiple Organ Dysfunction. However, the mechanism behind the onset of these extraneous symptoms following burn injury is not fully understood. The integrity of the gut barrier is of critical importance as it harbors the largest bacterial reservoir in the body. Following burn injury, we observed a breakdown of the gut barrier resulting in increased gut leakiness and bacterial translocation. Under homeostatic conditions, heat shock proteins (HSPs) stabilize tight junction proteins. Particularly, HSP72 is …

Tcr Modifications To Enhance Expression, Chain Pairing, And Antigen Recognition For Adoptive T Cell Transfer, Kendra Foley

Master's Theses

T cell receptor (TCR) gene modified T cells for adoptive T cell transfer therapy have been shown to have clinical success in treating melanoma and other malignancies by redirecting the specificity of peripheral blood lymphocytes (PBL) to recognize tumor and/or viral associated antigens of choice. One of the challenges in using TCR gene modified T cells is the potential mispairing between endogenous and introduced alpha and beta TCR chains, allowing for unwanted off-target reactivity, autoimmunity, or impaired therapeutic efficacy. One approach to augment proper TCR chain pairing and to enhance T cell function involves the modification of the introduced TCR …

Characterization Of Exopolysaccharide (Eps) Produced By Bacillus Subtilis Mutants, Alexander Argianas

Master's Theses

Bacillus subtilis is a probiotic bacterium that can protect against the murine-

specific attaching and effacing (A/E) pathogen Citrobacter rodentium. C. rodentium displays many features observed with the human pathogen enteropathogenic E. coli (EPEC). Protection by B. subtilis is mediated by exopolysaccharide (EPS), which is encoded by the epsA-O operon that contains 15 genes. Disruption of one of these genes, epsH, which appears to encode a putative glycosyltransferase, leads to the loss of protective capacity by B. subtilis and by EPS. EPS is a major component of the B. subtilis biofilm and the ΔepsH mutant is unable to form a …

Role Of The Intestinal Microbiota In Gut Barrier Dysfunction Following Burn Injury, Zachary Earley

Master's Theses

Burn injury represents a major medical problem with half a million cases requiring medical attention and 4,000 deaths reported annually. Sepsis and multiple organ failure remain the leading causes of death following injury, and may be brought on by bacterial infections or toxins. The gastrointestinal tract contains approx. 100 trillion microbes; therefore, the indigenous commensal microbiota may play a role in leading to these complications or infections in burn patients. The overall objective of this project is to identify a potential mechanism whereby changes in gut bacteria may lead to intestinal inflammation or bacterial translocation--key factors which may lead to …