MS, Kent State University, 2018, College of Arts and Sciences / School of Biomedical Sciences

Vaccine-induced, Th1-polarized immunity can have a positive impact on early HER-2pos breast cancer, especially when combined with drugs that block estrogen signaling. This suggests that pharmacological targeting of additional intracellular signaling pathways could also potentiate vaccine effects. We therefore tested the chief cytokine effectors of Th1 immunity (IFN-gamma; and TNF-a) in combination with targeted drugs including palbociclib (cyclin-dependent kinase 4/6 inhibitor) and sunitinib (broad spectrum receptor tyrosine kinase inhibitor) on a panel of human breast cancer cell lines (MCF-7, SKBR-3, JIMT-1, HCC-1419, MDA-MB-468 and MDA-MB-231). After multiday exposure, we examined the impact of combinations of these agents on a wide variety of cellular characteristics including metabolic activity (Alamar Blue assay), cell viability (Trypan Blue staining), loss of mitochondrial membrane potential (TMRE staining), and indicators of apoptotic cell death (Annexin V/propidium iodide staining, and cytochrome c release). The overall findings were that combined treatments with Th1 cytokines plus individual targeted drugs had significantly greater impact on breast cancer cell function than either cytokines or drug alone. These data suggest that palbociclib and sunitinib should be further investigated as possible adjuvants to vaccine therapy for breast cancer.

Committee: Gary Koski Ph.D (Advisor) Subjects: Biomedical Research

Master of Science, The Ohio State University, 2006, Graduate School

Committee: Not Provided (Other) Subjects:

Doctor of Philosophy, The Ohio State University, 2024, Animal Sciences

An estimated 70% of adults in the U.S. have experienced at least one traumatic event in their lives. Equine-assisted interactions (EAI) have been reported to contribute to positive outcomes for a variety of populations. However, little is known of the how previous trauma may influence these outcomes. EAI is a comprehensive term used to refer to a variety of services that utilize horses and other equine to benefit people. Although positive outcomes have been observed among EAI participants, research reports mixed results with few studies utilizing a multifaceted approach to assess responses. Additionally, few studies have sought to investigate the impact of EAI on the equine participant. Limited evidence suggests there is little to no effect of EAI on the equine involved but these studies lack a multifaceted approach which prevents a more comprehensive understanding of the equine experience. Therefore, the purpose of this study was to 1) assess the influence of equine assisted interactions (EAI) on human biopsychosocial responses, 2) assess the influence of previous trauma type on biopsychosocial responses to EAI, and 3) to assess the influence of EAI on equine participants using a biobehavioral approach. Participants (n = 36) interacted with the same equine (n = 10) in four consecutive, weekly 30-min EAI sessions. Human biological measures included heart rate (HR), heart rate variability (HRV), and salivary analytes oxytocin (OT), TNF- α, IL-1β, IL-6, and IL-8. Human psychological responses included participants' perceived quality of life, PTSD symptoms, momentary anxiety, and psychological distress. Human behavioral responses including standing, locomotion, sitting, and touch were assessed via scan sampling every 30 sec from video recorded every session. EAI sessions were video recorded to assess behavioral responses via scan sampling. Previous trauma was classified as TBI, PTSD, BOTH (PTSD/TBI), or NONE. Equine biological responses included heart rate (HR), heart (open full item for complete abstract)

Committee: Kimberly Cole (Advisor); Joseph Guada (Committee Member); Kathryn Lenz (Committee Member); Aubrey Fine (Committee Member) Subjects: Animal Sciences; Animals; Behavioral Sciences; Immunology; Psychobiology

Bachelor of Science (BS), Ohio University, 2023, Biological Sciences

Type 1 diabetes mellitus (T1D) is an autoimmune disease characterized by the loss of the only insulin secreting cells in the human body, the pancreatic beta-cells. Exogenous insulin is used to control the resulting hyperglycemia and replace the lack of insulin, but it is not a cure. To cure T1D, the autoimmune destruction of beta-cells must cease, and insulin secretory capacity must be restored. We have identified a novel compound that both protects against cytokine-induced cell death and stimulates insulin secretion from mouse islets. Importantly, I demonstrate that this compound exhibits protection from overnight cytokine treatment and enhances insulin secretion from human islets. Surprisingly, acute exposure to this compound severely reduced intracellular calcium. These findings conflict with observed effects on insulin secretion. However, the onset of enhanced insulin secretion appears between 2 and 4 hours with a maximal effect around 12 hours. In addition, the insulin secretory effects are shown to remain and enhance the response to glucose stimulation even after washout as treated islets showed increased insulin secretion in high glucose compared to control and glipizide-treated islets. We also found that our compound was one out of two compounds tested that augmented overnight insulin secretion. Testing of the amplifying pathway of insulin secretion revealed that this compound does enhance insulin secretion through this pathway. Finally, our lead experimental compound shows varying protection against ER stressors thapsigargin, brefeldin A, and tunicamycin. Overall, these findings suggest the small-molecule compounds we are developing and testing may hold therapeutic potential for treating type 1 diabetes through their dual action.

Committee: Craig S Nunemaker (Advisor) Subjects: Biology; Biomedical Research; Cellular Biology

Master of Science, Miami University, 2022, Microbiology

The signaling mechanism of SLAMF9 is not well defined but has been shown to promote proinflammatory cytokine production, which occurs through toll-like receptor (TLR) stimulation. To begin deducing the signaling mechanism, we looked at the potential of SLAMF9 positive cells to produce type-I IFNs and proinflammatory cytokines upon TLR stimulation in-vivo and in bone marrow-derived dendritic cells (BMDCs). Wild-type (WT) and Slamf9 knockout (KO) BMDCs were stimulated with different TLR agonists, to determine if SLAM9 positive cells require interaction with other cells types to affect proinflammatory cytokine production. We found that SLAMF9 does not have a cell-intrinsic effect on TLR-induced type-I interferon (IFN) production but might have a regulating role in TNF⍺ and CXCL1 production. We also looked at type-I IFN production from sorted SLAMF9 positive cells and negative cells from WT mice that were injected with CpG1585, TLR 9 agonist, to determine if SLAMF9 positive cells were major producers of type-I IFNs. We found that in both the liver and spleen that SLAMF9+ cells were the major producers of type-I IFNs. Overall, SLAMF9 does not have a strong effect on early type I IFN production, in vitro, but might regulate TNF⍺ and CXCL1.

Committee: Timothy Wilson (Advisor) Subjects: Immunology

Master of Science, The Ohio State University, 2022, Dentistry

Background and Objectives: The etiopathogenesis of peri-implant diseases is not well understood. Although dental plaque biofilm related bacteria may be the major player, the longterm integrity of implant devices made from titanium alloys and its effect on peri-implant pathogenesis have been recently questioned. Our group previously reported the presence of titanium ion (Ti) within peri-implant and adjacent gingival crevicular fluid (PICF and GCF) not only at sites diagnosed with peri-implant diseases but also with peri-implant health. We also published on peri-implant tissue response to surgical trauma following implant placement surgery by studying PICF and adjacent GCF cytokine content. The current study aims to evaluate inflammatory response and possible Ti contamination during early phases of peri-implant wound healing following surgical placement of a dental implant fixture via two different surgical protocols (one versus two stage). Materials and Methods: Patients scheduled to receive their first dental implant placement surgery at an edentulous site were recruited. Demographic information and clinical measurements of patient's periodontal status were documented at initial visit. Saliva, GCF, PICF and gingival biopsy samples were collected prior to (saliva, GCF, PICF) and during (gingival biopsy) surgery, 1 week, 4 weeks (saliva, GCF, PICF) and at 4 months (saliva, GCF, PICF from all sites, gingival biopsy only from implant sites placed with two stage protocol) post-operatively. The GCF and PICF samples were analyzed for inflammatory mediator content using V- plex assays and GCF, PICF, saliva samples for titanium release using inductively coupled plasma-mass spectrometry [ICP-MS]. Gingival biopsies were processed for H&E and IHC staining to study inflammatory infiltrates and macrophage phenotypes. Multivariable regression analysis was conducted to investigate possible associations between cytokine content, Ti contamination and possible clinical determinant (open full item for complete abstract)

Committee: Binnaz Leblebicioglu (Advisor); Dimitris Tatakis (Committee Member); Damian Lee (Committee Member) Subjects: Dentistry

Bachelor of Science (BS), Ohio University, 2022, Biological Sciences

Among the complex factors contributing to beta-cell failure is inflammation. Elevated levels of proinflammatory cytokines in obese individuals increases the risk of developing type 2 diabetes (T2D), and there is evidence that these low levels of circulating cytokines lead to pancreatic islet dysfunction in T2D-prone individuals. Iron and zinc are closely connected to both the inflammatory response and diabetes; however, the interrelation is unclear. This thesis investigates changes in iron and zinc distribution within beta-cells under conditions of low-grade inflammation. Analysis of gene microarray data revealed iron- and zinc-regulating genes in islets to be sensitive to cytokines modeling low-grade inflammation. Namely, Hamp, Steap4, and Lcn2 were strikingly upregulated in cytokine-treated islets, and these genes are all involved in cellular iron accumulation and/or retention. Beta-cells scanned by synchrotron X-ray fluorescence showed iron in discrete structures that were significantly smaller and more iron-dense in cytokine-treated beta-cells, consistent with the sequestering function of identified iron-associated genes. These data suggest a network of iron-regulating genes in beta-cells plays a role in sequestering iron in response to low-grade inflammation. Fluorescence microscopy indicated the labile zinc pool is significantly reduced in beta-cells treated with low doses of cytokines (48 hr) compared to control beta-cells. During acute cytokine exposure (0-15hr), zinc was released into the labile pool, followed by an abrupt reduction in detectible zinc. Additionally, 48 hr cytokine exposure induced significant redistribution of labile zinc characterized by the loss of zinc from ER. This thesis presents insight into the ongoing study of the beta-cell metallome, particularly under conditions of obesity-induced inflammation.

Committee: Craig S. Nunemaker (Advisor); Soichi Tanda (Advisor) Subjects: Biology; Biomedical Research; Cellular Biology

Master of Science (MS), Wright State University, 2021, Pharmacology and Toxicology

Cytokines play a pivotal role in regulating inflammation, which is a condition that makes the tissue vulnerable to different pathological and physiological conditions. Thus, how cytokines are regulated is an important area of study. Skin that receives ultraviolet B radiation (UVB), a major pro-oxidative stressor, results in the release of multiple cytokines and chemokines like tumor necrosis factor (TNF)-alpha and interleukin (IL)-8. Previous studies from our group and others have demonstrated synergistic release of TNF-alpha when UVB is combined with IL-1 or the lipid mediator Platelet-activating factor (PAF). Of interest, subcellular microvesicle particles (MVP) have been proposed to play an important role in intercellular communication. Moreover, UVB and PAF agonists cause MVP release in keratinocytes. Therefore, we believe that understanding the role of MVP in these inflammatory responses could be insightful for photosensitivity mechanisms and to suppress inflammation. The current study focuses on the combination of low concentrations of PAF agonist and UVB in-vitro and ex-vivo to observe potential synergism in the release of cytokines and MVP. We also studied the effects of acid sphingomyelinase (aSMase) inhibitor imipramine, for its ability to modulate both MVP and cytokine release. The application of aSMase inhibitor inhibited the synergistic response of MVP and cytokines allows us to conclude the potential involvement of MVP in the exaggerated response of cytokines from combining UVB and PAF. These studies have potential relevance in understanding abnormal skin reactions such as photosensitivity.

Committee: Jeffrey B. Travers M.D., Ph.D. (Advisor); Mike Kemp Ph.D. (Committee Member); Ji Chen Bihl M.D., Ph.D. (Committee Member) Subjects: Pharmacology

Master of Science, The Ohio State University, 2021, Dentistry

Introduction: Microorganisms normally colonize the periodontal tissues, forming a biofilm, and changes in this biofilm can lead to colonization by opportunistic pathogens and lead to periodontal disease. Although we know that orthodontic appliances increase plaque retention, there are limited reports in the literature on how tooth movement affects the subgingival biofilm. The pressure tension theory states that orthodontic force causes pressure and tension sites in the periodontium, where there is bone resorption occurring in the pressure sites and bone formation occurring in the tension sites. The cytokine profile is also affected, as well as the amount of blood flow. Currently there are no studies describing whether the changes that occur during orthodontic tooth movement affect the composition of the subgingival biofilm, especially over the course of comprehensive orthodontic treatment, which lasts between one to two years for most patients. Objectives: The aim of this study was to understand how orthodontic tooth movement affects subgingival biofilm development. The secondary aims were to examine whether there is any difference in subgingival biofilm and cytokine profile in anchor teeth vs. non-anchor teeth, and in pressure vs. tension sites. Methods: Nine healthy subjects between the ages of 13 and 19 who were treatment planned for 1st maxillary premolar extractions were followed for 8 to 12 months. At each regularly scheduled adjustment appointment (once every 4 to 6 weeks), subgingival plaque samples and gingival crevicular fluid samples were taken from the mesiofacial and distofacial sites of the upper right canine (pressure and tension sites) and upper right first/second molar (anchor sites). Probing pocket depths, plaque index, and gingival index were recorded for the same teeth. Bacteria present in the samples were identified using 16S sequencing (Illumina MiSeq). Cytokine analysis was completed using the V-PLEX Proinflammatory Panel 1 Human Kit. (open full item for complete abstract)

Committee: Purnima Kumar (Advisor); Shareef Dabdoub (Committee Member); Do-Gyoon Kim (Committee Member) Subjects: Dentistry

Master of Science, Miami University, 2020, Microbiology

SLAMF9 is a cell surface protein expressed on the surface of a wide variety of cells. It is a peculiar but compelling molecule due to its lack of a known receptor, ligand, or signaling adapter. While there is little known about this protein, there is evidence suggesting SLAMF9 enhances clearance of bacterial pathogens but makes the host more susceptible to viral pathogens. This dual phenotype is thought to be the result of inflammatory cytokine crosstalk. Stimulation of SLAMF9 Knockdown and Control THP-1 cells with lipopolysaccharide (LPS) mimicking an early response to infection resulted in higher proinflammatory cytokine production in cells with normal SLAMF9 expression. A proteomic analysis was performed to determine which proteins, other than cytokines, were differentially expressed between the SLAMF9 Knockdown and Control THP-1 cells. It was determined that the expression of SLAMF9 influences several components of Ras signaling, which is a complex network regulating proliferation, survival signals, and cytokine transcription. A combination of differential proteomics and a phosphosite analysis indicated other proteins of interest to pursue in future research. Determining new regulatory mechanisms may give rise to new potential drug targets to resolve inflammatory disease.

Committee: Timothy Wilson PhD (Advisor); Eileen Bridge PhD (Committee Chair); Xin Wang PhD (Committee Member) Subjects: Immunology; Microbiology

Master of Science (MS), Wright State University, 2020, Pharmacology and Toxicology

Ventilator associated pneumonia (VAP) is the most common nosocomial infection leading to increased mortality in ICU patients. The main goal of this project was to evaluate and analyze a number of different biomarkers of disease that may be involved in the transition to pneumonia in patients on ventilators. Patients in this study were in the ICU, required greater than or equal to 48 hours of mechanical ventilation, and had a Clinical Pulmonary Infection Score greater than or equal to 6. For this study we recruited 16 subjects who were on ventilator for more than 48 hours, obtained serum and sputum from each that was frozen at -80C and stored until use. Six patients developed pneumonia (VAP) and the rest were considered Non-VAP. Our goal was to analyze cytokine levels, Platelet-activating factor acteylhydrolase (PAFAh), procalcitonin, vitamin D, and pentraxins, in the sputum as well as serum samples from these patients. PAFAh is an enzyme that inactivates platelet activating factor, which is involved in cytokine release. Pentraxin3 is an early marker of lung injury. Using the Bio-Rad BioPlex system, we analyzed 27 cytokines in sputum and serum. Of these, we found that 6 were significantly increased in the sputum of VAP patients, while only 1 was significantly increased in the serum from VAP patients. The PAFAh assay revealed no significant difference between VAP and non-VAP patient sputum or serum, though there was a trend for lower PAFAh in VAP patients compared to non-VAP. Analysis for Pentraxin3 indicated that it was present in both sputum and serum though there was no significant difference. Sequencing of fragmented peptides from the sputum also revealed a peptide from pneumonia bacteria. In conclusion, the results strongly suggest that a larger study is needed, in addition to cytokines, other enzymes and proteins need to be explored as potential markers for early diagnosis and potential new treatments of VAP.

Committee: David R. Cool Ph.D. (Advisor); Ravi P. Sahu Ph.D. (Committee Member); Jeffery B. Travers M.D., Ph.D. (Committee Member) Subjects: Pharmacology

PHD, Kent State University, 2020, College of Arts and Sciences / Department of Biological Sciences

The dysregulation of pro-inflammatory cytokines as a result of stress exposure has become widely implicated in the complex etiology of depression. Patients with depression have been shown to have elevated levels of pro-inflammatory cytokines both in plasma and cerebral spinal fluid. During times of stress pro-inflammatory cytokines are primarily regulated by glucocorticoids and catecholamines that work in opposition to one another to inhibit (glucocorticoid receptor) or stimulate (beta-adrenergic receptor) pro-inflammatory cytokine production. While chronic stress can lead to heightened levels of both glucocorticoids and catecholamines, it is not known how the dysregulation of these systems affect the regulation of pro-inflammatory cytokines. This dissertation focused its work through three aims. Aim 1 explored norepinephrine's regulation of microglial pro-inflammatory cytokines. Aim 2 examined sex differences in the regulation of brain IL-1 beta by glucocorticoids and catecholamines following chronic stress. Finally, aim 3 sought to uncover glucocorticoids effect on norepinephrine turnover. Together, these aims help to better understand how chronic stress alters the regulation of brain pro-inflammatory cytokines with implications for depression.

Committee: John Johnson (Advisor) Subjects: Biology; Neurosciences

MS, University of Cincinnati, 2019, Allied Health Sciences: Nutrition

Objectives: To examine the relationship primarily between dietary intake of polyunsaturated fatty acids (total omega 6 and total omega 3 fatty acids), but also of fruits, vegetables, and selected vitamins and cytokines in children with asthma and allergic disease. Additionally, a preliminary assessment of the relationship between serum cytokine levels, intake of the dietary variables, and measures of disease control was explored. Methods: A secondary data analysis was conducted using data collected from the Biomarkers of Environmental Exposures in Asthmatic and Allergic Children study by researchers from Cincinnati Children's Hospital Medical Center. A total of 329 children, 5-18 years of age being treated for asthma or allergic disease were enrolled in the study. Data were collected and analyzed in two batches: batch 1 (n=65) in September 2010 and batch 2 (n=264) in December 2013. For each patient, a fasting blood sample (25-30mL) was collected and serum levels of IL-4, IL-6, IL-10, IL-13, IL-17 and TNF-a were measured. Subjects also completed a Block Food Frequency questionnaire (BFFQ), Asthma Control Test (ACT) and Respiratory Symptom Score test (RSS). For the ACT, higher scores indicate greater asthma control. While for the RSS, lower scores indicate greater asthma control. Multiple logistic regression analyses were conducted to determine the relationship between dietary variables and cytokine levels, as well as bivariate correlations to determine relationships between dietary variables and measures of disease control. Results: Cytokine data were significantly different between batches, and were therefore analyzed independently. There were no significant relationships observed between polyunsaturated fatty acid intake and serum cytokine levels, but both omega-3 and omega-6 intake were significantly, negatively associated with ACT score, (indicating that higher intake of these fatty acids was associated with lower disease control). In terms of significant (open full item for complete abstract)

Committee: Abigail Peairs Ph.D. (Committee Chair); Sarah Couch Ph.D. (Committee Member) Subjects: Nutrition

Master of Science (MS), Wright State University, 2019, Pharmacology and Toxicology

Microvesicle particles (MVPs) are subcellular particles that could be involved in inter-cellular communication because they carry various bioactive substances including cytokines. Previous studies from our lab has shown that that the lipid mediator Platelet-activating Factor (1-alkyl-2-acetyl-glycerophosphocholine; PAF) and ultraviolet B radiation (UVB) enhances the release of MVP in various cell types like primary keratinocytes, epithelial cell lines, and murine skin. We hypothesized that there may be synergistic increases in MVP release after combination of treatment of keratinocytes with UVB and PAF agonist (CPAF)/phorbol ester (PMA). The combination treatment significantly increases MVP and cytokine release at 4 to 8 hours time points. Imipramine, an acid sphingomyelinase (aSMase) inhibitor blocks MVP and cytokine release which indicates that MVP could be involved in cytokine release. The increased level of TNF-alpha in the supernatant was not present in the MVP but surprisingly there was increased level of anti-inflammatory cytokine, Interleukin 1- receptor antagonist (IL-1ra) inside MVP after UVB treatment than in combination treatment, which could portray potential imbalances between pro-inflammatory cytokines and down regulating responses in exaggerated condition. These studies suggest that MVP could play a role in how keratinocytes respond to UVB.

Committee: Jeffrey B. Travers M.D., Ph.D. (Advisor); Michael G. Kemp Ph.D. (Committee Member); Ji Chen Bihl M.D., Ph.D. (Committee Member) Subjects: Pharmacology

Doctor of Philosophy, The Ohio State University, 2018, Comparative and Veterinary Medicine

Neonates are immune competent but immunologically immature at birth and are highly susceptible to enteric pathogens. For example, porcine epidemic diarrhea virus (PEDV) is a highly virulent re-emerging enteric coronavirus that causes acute diarrhea, dehydration and 80-100% mortality in neonatal piglets. Since its emergence in the United States (US) in 2013, PEDV caused over 8.5 million piglet deaths and $900 million to $1.8 billion in losses to US swine producers in 2014. Lactogenic immunity remains the most promising and effective way to protect neonatal suckling piglets from enteric diseases like PEDV. This is particularly true for domesticated animals like swine and cattle whose epitheliochorial placenta inhibits immunoglobulin (Ig) transfer in utero. Therefore, colostrum [first secretions from the mammary gland (MG)] and milk-derived antibodies (Abs) and other immune factors are the sole source for immune protection after birth. Passive lactogenic immunity is achieved through high titers of IgG Abs in colostrum and a continuous supply of secretory IgA (sIgA) Abs in milk. Specifically, because of their persistence in milk at high titers, sIgA Abs play a major role in conferring passive lactogenic protection against enteric pathogens in suckling neonates. Therefore, our goal was to improve the understanding of maternal and lactogenic immunity induced via the gut-MG-sIgA axis and its role in protection against PEDV in neonatal suckling piglets. Our first objective was to study the impact of stage of gestation at time of PEDV infection in pregnant gilts and its implications for generation of lactogenic immunity and protection of PEDV-challenged piglets. To understand how stage of gestation affects maternal immune responses to PEDV, three groups of gilts were orally infected with PEDV in the first, second or third trimester. Control (mock) gilts were inoculated with medium in the third trimester. To determine if lactogenic immunity correlated with protection, all (open full item for complete abstract)

Committee: Linda Saif (Advisor); Anastasia Vlasova (Committee Member); Qiuhong Wang (Committee Member); Prosper Boyaka (Committee Member) Subjects: Agriculture; Animal Diseases; Immunology; Veterinary Services; Virology

PHD, Kent State University, 0, College of Education, Health and Human Services / School of Health Sciences

Heat illness is potentially life-threatening and poses an occupational hazard for firefights and military personnel who have to exert themselves in hot, humid environments. The purpose of this study was to examine how circulating concentrations of IL-6, IL-10, IL-1ra, and IL-1ß responded to temperature and humidity during aerobic exercise. Twelve college-age men completed six experimental trials: a VO2max test, and cycling in five conditions: 5ºC/passive humidity (LTLH), 22ºC/45% humidity (MTMH), 22ºC/70% humidity (MTHH), 35oC/20% humidity (HTLH), and 35oC/45% (HTMH). The environmental trials involved cycling for 60 minutes at 60% VO2max, resting for 15 minutes, cycling at 90% VO2max until exhaustion (TTE), then recovering for 60 minutes in the same temperature condition. Blood was obtained prior to exercise (PRE), after 60 minutes of cycling (60), after the TTE (90), and after recovery (REC). Blood was analyzed for serum levels of IL-6, IL-10, IL-1ra, and IL-1ß. There was a time effect for IL-6, IL-10, and IL-1ra in all conditions except LTLH. The increases in IL-6 were during exercise, at 60 and 90. For IL-10 and IL-1ra, the increases were at 90 and REC, respectively. The increases in IL-6 and IL-1ra were enhanced in HTLT and HTMH. For IL-1ß, there was only a time effect at LTLH. The IL-6 response indicates an inflammatory response to exercise that is enhanced in the heat. The delayed response of IL-10 and IL-1ra and the lack of response of IL-1ß in the heat, indicates that the exercise-induced anti-inflammatory response outweighs the pro-inflammatory response to heat.

Committee: Adam Jajtner (Committee Co-Chair); Ellen Glickman (Committee Co-Chair) Subjects: Environmental Health; Immunology; Physiology

Master of Science in Biological Sciences, Youngstown State University, 2018, Department of Biological Sciences and Chemistry

The goal of this study was to examine the host cytokine response to a mutant strain (ΔyakA) of T. marneffei. T. marneffei is a fungus that causes the disease talaromycosis in AIDS patients. The fungus is dimorphic and switches between growth forms depending upon temperature. T. marneffei exhibits mold growth at 25°C but transforms into yeast cells inside of the host which become intracellular pathogens, living inside a variety of cells including macrophages. Previously, our laboratory discovered a mutant, designated ΔyakA. The protein encoded by the yakA gene is a type of molecular sensor. Studies showed that the ΔyakA mutant of T. marneffei has a weakened cell wall, which may affect its pathogenicity. Therefore, conidia of the ΔyakA mutant were co-cultured with the human monocyte cell line THP-1 or the macrophage-like murine cell line J774 to investigate whether the weakened cell wall has an effect on cytokine production, since cytokines play a vital role in host response to infection. The levels of the pro-inflammatory cytokines TNF-α , IL-1β and IL-6 were measured from cell lines infected with the ΔyakA and wild-type conidia. The THP-1 human macrophages produced significantly more IL-6 and TNF-α when co-cultured with WT compared with ΔyakA but IL-1β showed no significant difference. There were no significant differences in cytokines measured from J774 mouse macrophages exposed to WT versus mutant conidia. Although some differences were noted between cytokine levels of macrophages incubated with WT versus ΔyakA conidia, the differences were not uniform enough to presently conclude that the yakA gene affects host cytokine response.

Committee: Chester Cooper PhD (Advisor); Diana Fagan PhD (Committee Member); Gary Walker PhD (Committee Member); Jonathan Caguiat PhD (Committee Member) Subjects: Biology; Immunology; Microbiology; Pathology

Master of Science, Miami University, 2018, Biology

To determine the role of spinal cord microglia following a cervical sympathetic trunk transection in the periphery, mice were fed a diet containing colony-stimulating factor-1 receptor (CSF-1R) inhibitor, which eliminated spinal cord microglia. Typical neuronal and glial plasticity and changes in cytokines were compared in mice fed CSF-1R inhibitor or control diet. While neuronal plasticity was unaffected, the baseline population of oligodendrocyte precursor cells (OPCs) was increased, and following injury, typical OPC plasticity was attenuated, with increased numbers of mature oligodendrocytes (OLs) observed. The elimination of microglia did not affect baseline astrocyte morphology or glial fibrillary acidic protein (GFAP) expression, yet an increase in injury-induced GFAP response was observed. Further, without microglia, the typical injury-induced increase in interleukin-1ß expression, a proinflammatory cytokine, was blunted and a cytokine screen revealed a decrease in several additional cytokines. These alterations reveal that microglia regulate activities of OL lineage cells and astrocytes in uninjured and injured animals and contribute either directly or indirectly to the cytokine profile expressed in the spinal cord following peripheral axon injury. These findings support the need to understand the complicated cross-talk between cell populations so that such interactions might be manipulated to promote neuronal survival following injury.

Committee: Lori Isaacson (Advisor) Subjects: Biology; Neurosciences

Master of Science (MS), Wright State University, 2017, Microbiology and Immunology

Allergy is a global health problem, affecting more than 40% of the world population. Allergy is often associated with a dysregulation of the immune system that becomes overactive against allergens. Several genes are known to be important for the proper homeostasis of the immune system in the epithelial tissues, and the loss of function of genes such as IKKß or SOX9 impacts the interaction between external stimuli and the host. Many of the allergic responses are triggered through the regulatory cascade governed by transcriptional factor NF-κB. Regulation of NF-κB activity is complex, involves several alternative routes, and is also influenced by metabolites produced in the gut by resident microbes. In this study, we aimed to profile small and large intestinal metabolite profiles in two mouse models of allergic disease, and compare these profiles to those of wild type mice. Proton nuclear magnetic resonance was used to acquire metabolite signatures of all samples. Multivariate statistical analyses of the binned NMR data revealed that samples separated into distinct groups corresponding to the gastrointestinal tract sections, and partially into groups based on mouse model. Using NMR spectrum deconvolution, we also quantified levels of twelve different metabolites known to be abundant in the gut lumen. Several quantified metabolites such as butyrate, propionate, tryptophan, and threonine were significantly less abundant in the gut of allergy disease mouse models compared to the wildtype mice. These metabolites were previously shown to regulate the signaling pathways of NF-κB. Knocked out the genes of IKKß or SOX9 in allergic mice models lowered the allergic responses in the airways by increasing the production of immunoglobulin A.

Committee: Oleg Paliy Ph.D. (Advisor); Nancy Bigley Ph.D. (Committee Member); Dawn Wooley Ph.D. (Committee Member) Subjects: Biochemistry; Immunology; Microbiology

Doctor of Philosophy (Ph.D.), Bowling Green State University, 2017, Biological Sciences

Polychlorinated biphenyls (PCBs) are environmental pollutants and endocrine disruptors, harmfully affecting reproductive, endocrine, neurological and immunological systems. This has implications for processes such as wound healing, which is modulated by the immunological response of the body. Conversely, while PCBs can be linked to diminished wound healing, outside of PCB pollution systems, exercise has been shown to accelerate wound healing. However, the potential for moderate intensity exercise to modulate or offset the harmful effects of a toxin like PCB are yet unknown. Exploration of this possible moderation on local immune response was achieved by measuring wound size and analyzing the concentrations of proinflammatory cytokines, interleukin-1ß (IL-1ß), interleukin-6 (IL-6), keratinocyte chemoattractant (KC), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor-a (TNF-a) in wounds (inflicted by punch biopsy) in mice that were not exercised as compared with those previously exercised at moderate intensity by running on a treadmill for 30min/day and then injected intraperitoneally with Aroclor 1254 (industrial mixture of PCB congeners) in doses of either 0, 100, 500 and 1000 ppm (wt/wt of mice). Mice were euthanized at Day 3 or Day 5 (n = 3-6) and skin excised from the wound area was homogenized and analyzed for cytokine content. Systemic effects of exercise on immune function in PCB exposed animals were examined by lipopolysaccharide (LPS) challenge (intraperitoneal injections) and analyzed by measuring the average body temperatures using a thermal imaging camera. Wound healing data revealed that in animals not exercised only the greatest dose of PCB (1000 µg/g) showed a pattern for faster wound healing. Exercise produced a pattern of more rapid wound healing rates compared to the animals administerd similar doses, except for animals administered 100 µg/g PCB. Concentrations of pro-inflammatory cytokines revealed patterns t (open full item for complete abstract)

Committee: Lee Meserve Dr. (Advisor); Howard Cromwell Dr. (Other); Todd Keylock Dr. (Committee Member); Scott Rogers Dr. (Committee Member); Vipaporn Phuntumart Dr. (Committee Member) Subjects: Immunology; Kinesiology; Molecular Biology; Physiology; Toxicology