Master of Science, The Ohio State University, 2010, Computer Science and Engineering

Today's imaging technologies generate a wealth of data that requires visualization and multi-dimensional quantitative image analysis as prerequisites to turning qualitative data into quantitative values. Such quantitative data provides the basis for mathematical modeling of protein kinetics and biochemical signaling networks that, in turn, open the way toward a quantitative view of cell biology. There has been a lot of work on solving some of the fundamental problems in the image based analysis like segmentation, image restoration, shape normalization etc. One of the limiting factors of such analysis is the statistical and morphological variation present across different biological samples. Thus, these standalone solutions can not be used off the shelf. The challenge is to develop a workflow for each problem considering the goals of the study, underlying biology and technical limitations. During pregnancy, the antibodies are transferred from mother to the fetus. This transfer takes place through placenta. It is well established that a MHC-1 receptor FcRn in essential for transport of IgG across placenta. But the exact mechanism of this process is not known. This thesis focuses on designing a workflow based on 3D analysis of confocal microscopy images and statistical modeling to understand the IgG transport in the yolk sac endoderm of the mouse. The workflow consists of image correction, active contour based segmentation, distance transform based protein distribution and clustering.

Committee: Raghu Machiraju PhD (Advisor); Dr. Clark Anderson MD (Committee Member) Subjects: Biomedical Research; Computer Science

Master of Science, The Ohio State University, 2024, Immunology and Microbial Pathogenesis

Nontuberculous mycobacteria (NTM) are an increasingly common cause of respiratory infection in people with cystic fibrosis (PwCF). Relative to those with CF and no history of NTM infection (CF-NTMNEG), PwCF and a history of NTM infection (CF-NTMPOS) are more likely to develop severe lung disease, experience complications over the course of treatment and are precluded from lung transplant eligibility. We recently reported lung lobe-specific immune profiles of 3 cohorts (CF-NTMNEG, CF-NTMPOS and non-CF adults) and found that the CF-NTMPOS airways are distinguished by accumulations of B cells produced. Confirming the presence of antibodies, in the bronchioalveolar fluid (BAL), would prove a mechanistic connection between B cell abundance and lung tissue damage. This would be due to an overreactive antibody response or the abundance of antibodies being able to exacerbate pulmonary damage. This is caused by the release of cytokines and other inflammatory signals to recruit more immune cells to the site. We anticipate these findings will lead to the development of a clinical diagnosis of NTM infection and the creation of host-directed therapies to improve NTM disease treatment in PwCF. We hypothesize that the presence of total antibodies in BAL for these three cohorts will provide a mechanistic connection to the exacerbation of lung tissue damage in PwCF and a concurrent NTM infection.

Committee: Richard Robinson PhD (Advisor); Emily Hemann PhD (Committee Member); Adriana Forerro PhD (Committee Member); Mark Drew PhD (Advisor) Subjects: Health Care; Immunology; Microbiology; Molecular Biology

Master of Sciences, Case Western Reserve University, 2024, Pathology

Malaria caused by the Plasmodium vivax parasite is a leading cause of disease burden in many tropical and subtropical parts of the world. Infection leads to the generation of antibodies specific to parasite proteins, such as apical membrane antigen 1 (AMA1). This ligand plays a role in invasion of reticulocytes by binding of the rhoptry neck (RON) complex, which is composed of RON proteins. This complex is inserted by the parasite into the cell surface. In malaria infections, IgG1 and IgG3 isotype frequencies are increased. These isotypes can activate complement or bind Fc receptors to initiate effector functions. As part of a project that isolates cDNA from individual PvAMA1- specific IgG expressing B cells, we aim to develop a rapid method to identify the IgG subclass from these B cells to provide a more comprehensive view of immunoglobulin generation in these individuals.

Committee: Neena Singh (Committee Chair); Jürgen Bosch (Committee Member); Christopher King (Committee Member) Subjects: Immunology; Molecular Biology; Parasitology; Pathology

Master of Science in Biomedical Sciences (MSBS), University of Toledo, 2022, Biomedical Sciences (Medical Microbiology and Immunology)

Burkholderia pseudomallei is a Gram-negative bacterium that causes the severe febrile disease melioidosis. The wide variety of signs and symptoms associated with melioidosis hinders timely diagnosis of this illness, which often allows the infection to progress to fatal septicemia before this bacterium is accurately identified. Successful treatment of melioidosis patients is further complicated by the extensive repertoire of antibiotic resistance mechanisms employed by B. pseudomallei. Due to the low dose required to cause fatal infection and the difficult nature of properly diagnosing and treating melioidosis, B. pseudomallei has great potential for deliberate misuse in acts of bioterrorism and is therefore classified a Tier 1 select agent by the Centers for Disease Control and Prevention. Thus, there is great interest in finding therapeutic targets to prevent and/or treat melioidosis, as no FDA-approved vaccine currently exists. Previous studies from our lab and others demonstrated that B. pseudomallei must evade the complement system in order to successfully persist within the host. More recently, unpublished work from our lab demonstrated that B. pseudomallei binds Factor H, the host's complement regulatory protein, to improve B. pseudomallei survival within the host. Importantly, the proteins involved in this immune evasion strategy are considered to function as effective therapeutic targets, as loss of binding Factor H may incur a fitness cost to the pathogen. Preventing the interaction between B. pseudomallei and Factor H may enhance serum susceptibility and/or bacterial clearance by host phagocytic cells. A novel and promising therapeutic approach for serum-resistant pathogens is the recombinant fusion protein Factor H-Fc. These chimeric constructs are composed of either Factor H domains 6-7 or 18-20 linked to constant regions (Fc) of immunoglobulin G isotypes 1 or 3 (Fc1 and Fc3, respectively). Potential mechanisms for how these might act as a therapeutic (open full item for complete abstract)

Committee: Mark Wooten (Advisor); Viviana Ferreira (Committee Member); Robert Blumenthal (Committee Chair) Subjects: Biomedical Research; Health Sciences; Immunology; Microbiology

PHD, Kent State University, 2015, College of Arts and Sciences / School of Biomedical Sciences

Many types of epilepsy, autoimmune or otherwise, are associated with the presence of autoantibodies against neuronal proteins. Paradoxically, antibodies (IVIg) have also been used to treat epilepsy. The goals of this research were twofold: 1) Determine the CNS location of antibodies in patients with non-autoimmune epilepsies and the targets of these antibodies; and 2) Examine the effects of endogenous and exogenous specific and non-specific antibodies in two status epilepticus (SE) models Immunohistochemistry and Western blotting were used to localize antibodies in patients with epilepsy, multiple sclerosis (MS) and arteriovenous malformation. Further analysis by ELISA, HEp-2 assay and immunoprecipitation revealed antibody targets. In mouse model experiments, lupus-prone or C57B6/J mice were injected with pilocarpine or kainic acid and monitored by EEG. Mice were treated with IV or IP injection of native or denatured IgGs, at time of or 12 hours before chemoconvulsant. Tissues were processed for immunohistochemistry and ELISA. Brain regions from patients with epilepsy contained extravasated IgGs. Intracellular antibodies were found in epilepsy but not in MS brain. In brain from patients with epilepsy, only neurons displayed nuclear IgGs. All subcellular fractions from brain resections of patients with epilepsy contained extravasated IgGs. In the nuclear IgG pool, anti-histone autoantibodies were identified by two independent methods. Serum analysis revealed anti-histone and anti-chromatin antibodies only in patients with epilepsy. In lupus-prone mice elevated serum IgGs favored post-SE survival. C57B6/J mice injected with native rat IgGs displayed a 40% reduction in pilocarpine-SE compared to control. IgGs extravasated in brains of untreated SE mice, but IgG-treated mice, with no pilocarpine-SE, experienced no parenchymal accumulation of IgGs. IgG leakage was observed in brain samples from KA treated mice and IgG treatment was largely ineffective. These r (open full item for complete abstract)

Committee: Derek Damron Ph.D (Committee Chair); Trine Jorgensen Ph.D (Committee Member); Gary Koski Ph.D (Committee Member); Ernest Freeman Ph.D (Committee Member) Subjects: Biology; Biomedical Research; Cellular Biology; Immunology; Medicine; Neurobiology; Neurosciences

Doctor of Philosophy (PhD), Ohio University, 2015, Physics and Astronomy (Arts and Sciences)

The force dependence of receptor–ligand bonding has been extensively studied in recent years. Many ultrasensitive force techniques have been used to measure receptor-ligand bonding interaction in biophysics. Microcantilevers have easily tunable and variable spring constants over other techniques. This work presents in detail the measurement methods of glass fiber microcantilever spring constants. Microcantilevers made from E-glass fibers were mounted in viewing chambers and imaged under a microscope. Multiple sets of images for each microcantilever were captured and analyzed using custom LabVIEW IMAQ image processing program with the centroid tracking images processing. The centroid data were processed using a 20 point moving average filter which was the optimum number of average points in a moving average filter. The results of microcantilever spring constants from thermal fluctuation method measurement were compared with elasticity theory calculation. The measured spring constants of 3 - 7 mm-long fibers were within the relative error expected for the spring constant due to the fiber length and diameter uncertainties. A novel forced unbinding system was developed for the receptor-ligand binding experiments. The system applied hydrodynamic flow from one micropipette to a glass fiber microcantilever tip which was brought into contact with a bead held in another micropipette with suction pressure. The displacement of the fiber tip increased linearly as the water pressure difference increased which is the magnitude of applied pressure used to generate flow from a micropipette. The interaction of protein A and human IgG was measured using the novel forced unbinding system. Heat treatment of the Bovine Serum Albumin is necessary to reduce the frequency of nonspecific adhesion. The percentage of adhesive events decreased from 2.69 % to 0.57 % after the BSA was denatured. The protein A and human IgG binding interaction was investigated for specific adhesion experiments (open full item for complete abstract)

Committee: David Tees (Advisor) Subjects: Biophysics

Master of Science (MS), Wright State University, 2006, Pharmacology and Toxicology

The present study was conducted to determine the half-life, assess the toxicity, and passive protection efficacy of purified immunoglobulin G (IgG) from Anthrax Vaccine Adsorbed (AVA) vaccinated human donors. Half-life determinations were calculated from the reportable values obtained using the anti-PA ELISA assay and the Centers for Disease Control's (CDC) “ELISA for Windows” software. For toxicity evaluations animals were observed clinical for one hour post administration and for 14-days post-treatment. The protection efficacy was determined based upon the mortality results from a lethal Bacillus anthracis aerosol challenge. While no protection was achieved in this delayed exposure scenario, the study yielded valuable kinetics data for use in future research.

Committee: Robert Casillas (Advisor) Subjects: Biology, General

MS, University of Cincinnati, 2012, Medicine: Immunology

Anaphylaxis is an acute-onset and potentially fatal systemic allergic reaction affecting proximately 0.05 to 2% of the population worldwide and can be induced by a wide range of allergens from foods, medications and insect venoms. Although IgE is commonly involved in human anaphylaxis, other mediators, such as non-IgE antibodies and complement are also reported to be involved. Thus we sought to evaluate the role of complement in IgE-independent anaphylaxis. To approach this, we have studied an IgG2a-mediated passive systemic anaphylaxis model in mice with different backgrounds and gene deficiencies. We demonstrate that complement component 3 (C3) plays a major role in anaphylaxis in C57BL/6 and Balb/c mice sensitized with a low dose of IgG2aa mAb Hy1.2 but not in mice sensitized a higher dose of Hy1.2, and that C5 is not involved in IgG2a-mediated anaphylaxis.

Committee: Fred Finkelman MD (Committee Chair); David Hildeman PhD (Committee Member); Nives Zimmermann PhD (Committee Member) Subjects: Immunology