Immunology and Infectious Disease Commons^™

Examining The Ligands, Location, And Function Of Toll-Like Receptor 10, Kiefer Keanu Tam

Master's Theses

Toll-like receptors are a class of innate immune pattern recognition receptors that detect pathogen-associated molecular patterns and regulate downstream inflammatory responses. Toll-like receptor 10 remains the only human TLR whose ligand specificity, cellular localization, and immunological function are not fully defined. Emerging evidence suggests that TLR10 may play an anti-inflammatory role and can heterodimerize with TLR2 to modulate immune signaling. Previous studies from our laboratory demonstrated that stimulation with heat-killed Mycobacterium tuberculosis promotes TLR2-TLR10 heterodimerization and trafficking to the cell surface. In this study, we expanded our ligand panel to include additional gram-positive bacteria, particularly anti-inflammatory probiotic species, to further …

Stability Insights From Modeling Chronic Myelogenous Leukemia, Giovani Thai

Master's Theses

This thesis centers around a model for chronic myelogenous leukemia (CML) as it behaves under imatinib treatment, a common medication for CML patients, and the anti-leukemia immune response. The dynamics are represented with a system of nonlinear delay-differential equations first constructed by Kim et al. in 2008, capturing population changes of T-cells and various CML growth stages. We investigate stability in both the clinical and mathematical sense. Through numerical simulations, we computationally incorporate a supplementary treatment plan to determine its effectiveness in aiding immune response and medication in achieving remission and full elimination. The primary goal is to conduct a …

Impact Of Temperature And Microbial Infection On The Honey Bee Hemocytes, Michael E. Oeth

Master's Theses

Introduction

Honey bees (Apis mellifera) rely on hemocytes for immunity, wound healing, and hemolymph homeostasis. While 32°C reflects optimal hive conditions, suboptimal temperatures (22°C) and stressors like pathogens (Varroa destructor, bacteria) or pesticides impair hemocyte function. Using clodronate liposomes (CLD) to deplete hemocytes, we assessed how immune suppression interacts with these stressors.

Methods

Newly emerged (5-day-old) and older (15-day-old) bees were assigned to: CLD, control liposomes (LP), PBS, injection wound, or no treatment. Cohorts were exposed to 32°C (optimal) or 22°C (stressful for non-immunosuppressed bees). Fifteen-day-old bees were challenged with Escherichia coli or Staphylococcus aureus. Survival, hemolymph volume, and hemocyte …

Lack Of Inflammatory Response In Mouse Embryonic Cells To Listeria Infection, Damilola Oyebode

Master's Theses

Approximately 1 in 5 pregnancies in the United States results in miscarriage, with a significant percentage of these losses attributed to failed implantation due to immunological challenges encountered by the early embryo. Recent research has shown that key cell types within mouse blastocysts, including embryonic stem cells (ESCs) and trophoblast stem cells (TSCs), exhibit remarkable resilience against the cytotoxic effects associated with viral infections and embryotoxic cytokines, such as TNFα and IFNγ, both of which are known to hinder blastocyst development. However, the immune properties of the preimplantation stage embryo, specifically the blastocyst, remain poorly understood. Using mouse cell model, …

Testing Strategies To Develop A Mucosal T Cell Inducing Vaccine For Hiv-1, Reynolds Boone

Master's Theses

Despite numerous efforts, HIV vaccine is still not available. Because HIV Envelop protein varies among subtypes and isolates, it is impossible for one vaccine against HIV Env to induce neutralizing antibodies against all subtypes and isolates. Previous vaccine attempts inducing HIV-specific CD4+ T cells have even been shown to enhance the HIV infection because HIV preferentially infects activated CD4+ T cells. It is known that CD8+ cytotoxic T cells (CTLs) play an important role in controlling HIV infections. We hypothesize that a vaccine inducing only CD8+ CTLs but not CD4+ T cells will be effective in preventing HIV infections. Thus, …

The Anti-Inflammatory Properties Of Exopolysaccharide From Bacillus Subtilis, Nikolay Todorov

Master's Theses

Commensal microorganisms can be used therapeutically to ameliorate a variety of diseases. The laboratory of Dr. Katherine L. Knight studies these therapeutic properties in the commensal microorganism known as Bacillus subtilis. Previously, the Knight lab has demonstrated that B. subtilis can protect against colitis caused by Citrobacter rodentium suggesting that B. subtilis possesses anti-inflammatory properties. These properties are specific to the polysaccharide capsule of the microorganism known as exopolysaccharide or EPS. EPS can be produced and purified for study. EPS has been demonstrated to be protective in a variety of diseases such as allergic eosinophilia, sepsis, Graft versus Host disease, …

Characterization Of Cd11c-Expressing Gut-Associated T Cells, Ana Cvetkovic

Master's Theses

Our immune system is a remarkable defense network that protects our bodies from a range of threats. Its’ intricate machinery comprises of messenger and effector cells that work together to coordinate an effective immune response. Traditionally, T lymphocytes have been considered effectors: they receive instructions from messengers, known as Antigen Presenting Cells (APCs), that alter T-cell activation state and behavior. In the gut, Dendritic Cells (DCs) are the APCs that most often instruct T cells. To accomplish their messenger role, DCs express CD11c at high levels, a surface marker involved in cell-to-cell signaling and phagocytosis. As such, CD11c expression has …

Impact Of Toll-Like Receptor 10 Mutations On Tuberculosis Infections, Adriana Nicole Rafael, Sangman Kim

Master's Theses

Tuberculosis (TB), caused by the intracellular pathogen Mycobacterium tuberculosis (M. tb), is a leading global health crisis causing millions of cases and deaths each year. Though it affects individuals of any race, ethnicity, or cultural group, research shows the disease disproportionately affects individuals with Asian ancestry. Immunity to M. tb starts with its recognition by pattern recognition receptors such as Toll-like receptors (TLRs), which are key players in immune responses. TLR-10, an understudied toll-like receptor due to its inability to be studied through mouse models, is thought to play a key role in the clearance of inflammatory diseases. SNP I775V, …

The Role Of B Cell Activation State And Sex In Aryl Hydrocarbon Receptor Mediated Induction Of Chemokine Receptor 9 And Alpha4beta7 Expression In Vitro, Logan Bauerle

Master's Theses

Defense of mucosal tissues from microbial infection and allergy is reliant on continual production of antibodies. The aryl hydrocarbon receptor (AhR) is known to regulate B cell development and is associated with suppression of systemic humoral immunity. Recent attention has been paid to the role of the AhR in altering expression of cell adhesion molecules (CAMs). B cells express CAMs and chemokine receptors to migrate around the body for localized secretion of antibodies. AhR agonists promote B cell migration to the small intestine through upregulation of chemokine receptor 9 (CCR9) and integrin α₄β₇. Both the AhR …

Role Of Toll-Like Receptor 2 In Macrophage Recognition And Response To Borrelia Burgdorferi, Yukiye A. Koide

Master's Theses

Lyme disease is the most common vector-borne disease in the US and will experience an uptick as the insect host, the ixodid tick, gains habitat with climate change. The causative agent of Lyme disease, Borrelia burgdorferi, is recognized by Toll-like receptor 2 (TLR2), which initiates the innate immune response. Here, I used ELISA and SEAP detection assays to determine that knocking out TLR2 in THP-1 macrophages reduces the amount of NFkB activation, as well as IL-10 and IL-1β secretion. Then, I used fluorescence microscopy and Incucyte assays to quantify the amount of phagocytosis performed by wild-type and knock-out THP-1 cells. …

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 …

Lactobacillus Metabolite Nc As An Inflammation Inhibitor, Drew Hahnnah Landrowski

Master's Theses

The human body contains a diverse ecosystem of microbes. Commensal bacteria have numerous mutually-beneficial relationships with their host. Not only do they assist in digestion and gut homeostasis, they are critical for development and function of the immune system. As such, hosts and microbes alike have evolved mechanisms to foster symbiosis. Gut barrier function, toll-like receptor signaling, and metabolites are all examples of how hosts interact with microbes to promote this mutually-beneficial relationship. Metabolites from commensal bacteria promote the generation of regulatory T-cells, capable of ameliorating inflammation. A number of inflammatory, allergic, and autoimmune conditions are associated with dysbiosis. Therefore, …

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 …

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 …

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 …

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 …

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 …

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 …

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 …

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 …