Immunology and Infectious Disease Commons^™

Host Pathogen Interactions: Is Arabidopsis Thaliana Remembered By Its Nemesis Pseudomonas Syringae?, Daniel Z. Kreiser

Lawrence University Honors Projects

Plants contain innate immune systems that deter pathogen infection. Pattern recognition receptors bind microbe-associated molecular patterns (MAMPs), triggering immunity. MAMPs are proteins exclusive to pathogens that are typically indispensable for their survival. For this reason, MAMPs cannot be mutated or removed without causing pathogen death. However, this does not necessitate constitutive expression of MAMPs. In this study, the MAMP response of Arabidopsis thaliana was utilized to determine differential detection of MAMPs expressed by Pseudomonas syringe pv. tomato DC3000 when pretreated with A. thaliana. Results demonstrated that more MAMPs are detected when P. syringae had previously encountered A. thaliana, …

Cell-Mediated Immunity In Porcine Reproductive And Respiratory Syndrome Virus, Rajeshwari Parida

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Porcine reproductive and respiratory syndrome virus is a significant swine pathogen which exhibits considerable sequence diversity. In an attempt to identify highly conserved T-cell epitopes contained in proteins of this virus, we examined heptadecamer peptides spanning the sequence of the PRRSV nonstructural proteins 9, 10 and 11, all of them are highly conserved, for their ability to elicit a recall proliferative and interferon-gamma response in peripheral blood mononuclear cells obtained from pigs immunized against the type-II PRRSV strain FL-12. These studies led to the identification of seven peptides, two from each NSP 9 and NSP 10 and, three from NSP …

Cloning And Characterization Of The Cell Wall Acting Enzyme Cd1034 From The Pathogen Clostridium Difficile, Zachary Suter

Honors Projects in Science and Technology

The manifestation of multidrug resistance in bacteria over the past several decades has resulted in one of the foremost challenges in the management of infectious diseases. The question arises, “How do we address this growing problem?” One solution to stem the growing rise in antimicrobial resistance is to investigate new targets, while another approach is to re-examine classical antibacterial targets with a fresh perspective. The aim of this paper is to begin the process of antibacterial development for the pathogen Clostridium difficile by characterizing the cell wall acting glucosaminidase CD1034. It is inunderstanding how CD1034 functions biochemically that it can …

Parasites Of Civets (Mammalia, Viverridae) In Sabah, Borneo: A Coprological Survey, Christina P. Colon, Sharon Patton

Publications and Research

Parasite data were collected from unlogged and selectively logged lowland rain forests in Borneo. Scat from a female Viverra tangalunga (Malay civet) in the unlogged forest contained eggs of Capillaria spp., Trichuris sp., Mamomoganomus sp., and nematode larvae. Scat attributed to V. tangalunga contained reproductive products (eggs/oocysts/cysts/larvae) of Capillaria sp., Toxocara sp., Ancylostoma sp. (probably A. ceylanicum), Isospora spp., strongylate-type eggs (probably Ancylostoma spp.), Paragoni­ mus sp. and other Trematoda eggs, Monocystis sp. cysts, Gnathostoma sp. (probably G. spinigerum), the lungworm Viverrostrongylus brauni, pinworm-type eggs, Eimeria spp., Sarcocystis-type sporocyst, dermanysid mites and mesostigmatid mites. Parasites from gut samples from two …

Thoughts On Quorum Sensing And Fungal Dimorphism, Kenneth W. Nickerson, Audrey L. Atkin, Jessica C. Hargarten, Ruvini U. Pathirana, Sahar Hasim

Papers in Microbiology

Farnesol has been best studied for its role in regulating fungal dimorphism. However, farnesol is also a lipid and in this review we analyze data relevant to farnesol’s function and synthesis from the perspective of farnesol and bacterial endotoxins acting as membrane active compounds. This analysis implicates the possible roles of: (1) endotoxins in the regulation of farnesol production by C. albicans; (2) farnesol in the interactions between C. albicans and the host during disseminated infections; and (3) ubiquinones in the mechanisms for unusually high resistance to farnesol by some C. albicans cell types. Finally we discuss the implications …

Therapeutic Peptide-Based Vaccination Strategies Against Hpv-Induced Cancers, Kelly Barrios Marrugo

USF Tampa Graduate Theses and Dissertations

There is an urgent need for the development of an effective therapeutic vaccine against cancer caused by human papilloma virus (HPV). We focused on HPV-induced malignancies because of their high worldwide prevalence (e.g., cervical carcinoma and head & neck cancer). A successful therapeutic vaccine could prevent the 250 000 deaths/year worldwide and the 2.25 billion dollars that

are expended in related care in the US.

We used an HPV-induced mouse cancer model to test vaccines

composed of a CD8 T cell peptide epitope administered with potent adjuvants designed to generate vast numbers of high avidity cytotoxic T lymphocytes specific for …

Examining Autoimmune Sequelae During Cancer Immunotherapy In A Combined Autoimmune Thyroiditis And Tumor Mouse Model, Suresh Kari

Wayne State University Dissertations

Some clinical trials of immunotherapeutic agents against cancers have resulted in the development of autoimmune sequelae, including autoimmune thyroiditis. It has been suggested that the immunotherapy resulted in an alteration in the immunoregulatory mechanism(s). Using experimental autoimmune thyroiditis (EAT), a mouse model of Hashimoto's thyroiditis (HT), the importance of regulatory T cells (Tregs) has been well established; inhibition of Tregs leads to enhanced thyroiditis with repeated doses of mouse thyroglobulin (mTg) without adjuvant. To simulate cancer patients, we developed a combined EAT and tumor model in our EAT-susceptible mice (CBA/J). In first establishing the tumor model, we found Treg depletion …

Membrane-Bound Immunomodulators As Adjuvants In A Cell Culture-Based Avian Influenza Vaccine, David Daniel Fischer

Wayne State University Dissertations

Inactivated viral vaccines often generate suboptimal immune responses. Adjuvants are incorporated into vaccines to increase their immunogenicity, however currently available adjuvants have shortcomings which have limited their use in human and veterinary medicine. This necessitates the development of new adjuvants and delivery systems. Cytokines have been extensively tested as adjuvants in vaccines but challenges such as diffusion from antigen, short half-lives and production costs have been encountered. To address this, we developed a technology that efficiently produces inactivated, whole-virus influenza vaccine bearing membrane-bound cytokines. Tethering the cytokine to the antigen of interest keeps the immunomodulator in close contact with the …

Establishment Of A Quiescent Infection Of Hsv-1 In L929 Fibroblasts Using A Mitotic Inhibitor And Ifn-Γ, Neelam V. Shinde

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The goal for this study was to determine if a quiescent infection of HSV-1 could be induced in murine fibroblasts L929 by treating them with the anti-mitotic agent 5-fluoro 2'deoxy uridine (FUDR) alone and with interferon-γ. Since neurons are post-mitotic and exhibit a lower metabolic rate than other cells, fibroblasts were treated with FUDR to induce a post-mitotic state. The cell cycle arrest of fibroblasts would decrease the thymidylate metabolism and impair HSV-1 replication. An evaluation of cytopathic effects of FUDR was used to determine the optimal concentration which arrests cell growth and inhibits viral replication. Image J program developed …

Q-Ve-Oph, A Control Caspase Inhibitor For Analyzing Neuronal Death, Rebecca L. Bricker

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Sarin is a neurotoxin that has been used in terrorist attacks in Japan and is a potential bioterrorist weapon. It induces seizures by affecting the regulation of neurotransmitters in the brain. Seizures are directly correlated to neuronal damage. Two types of neuronal damage that can occur are apoptosis and necrosis. One of the main regions of the brain where neuronal death occurs is the hippocampus, which is involved in memory. Victims showed chronic decline of memory loss 3 and 7 years after the Tokyo's terrorist attack incident. There are treatments available that can break down sarin or can block the …

The Effects Of Enteropathogenic And Commensal Escherichia Coli On Tight Junction Permeability, Hilary Kaye Allen

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The intestinal mucosa maintains a barrier between materials from the external environment and the internal environment of the host. Disruption of the gut wall integrity is involved in the development of various intestinal diseases, such as Irritable Bowel Syndrome (IBS) and Crohn's disease. The intestinal mucosa is lined with epithelial cells that are connected by tight junctions, the intercellular junctions that form a selectively permeable barrier between paracellular pathways. Enteric pathogens, such as enteropathogenic Escherichia coli (EPEC), can disrupt the tight junctions of epithelial cells by altering the cellular cytoskeleton or by directly affecting tight junction proteins. Commensal Escherichia coli …

Mhc Class I Expression In Murine Fibroblast And Keratinocyte Cell Lines During The First Twenty-Four Hours Of Infection With Herpes Simplex Virus-1 (Hsv-1), Prasanthi Kumchala

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The hypothesis of this study is: HSV-1 infection of murine fibroblasts and keratinocytes inhibits expression of MHC class I molecules during first 24 hours of infection. IFN-γ pretreatment of fibroblasts protected the cells from virus-induced inhibition of MHC class I expression, but did not protect keratinocytes. Herpesviruses are known for their ability to establish persistent infections. Herpesviruses exert many different ways to suppress host defense mechanisms. One such way is by down regulating expression of the major histocompatibility complex I (MHC I) molecules in infected cells. Epidermal cells such as keratinocytes are the major sites for herpes simplex virus type …

The Effects Of Hsv-1 Challenge On Polarized Murine Macrophages: An In Vitro Model Using The J774a.1 Murine Macrophage Cell Line, Adam Craig Reichard

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In our current study we examined the effects of HSV-1 challenge on J774A.1 macrophages polarized to either a proinflammatory (M1) or anti-inflammatory (M2) phenotype. Polarized J774A.1 macrophages were characterized using CD14-CD86 and SOCS1-SOCS3 expression levels. SOCS proteins are a family of proteins that are capable of inhibiting cytokine-signaling pathways. HSV-1 up regulates expression of SOCS1 protein levels in infected cells, inhibiting the ability of infected cells to produce proinflammatory products (Nowoslawski Akhtar and Benveniste, 2011). This study shows that signals within the microenvironment play a greater role in macrophage polarization, and SOCS1-SOCS3 expression levels, than does HSV-1 challenge. M1 macrophages …

Suppressor Of Cytokine Signaling (Socs) 1 & 3 Expression In Hsv-1- Infected And Interferon-Γ-Treated Neuro-2a Cells, Melinda Jones

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This study examined the effects of HSV-1 infection and IFN-γ treatment on Neuro-2A cells. HSV-1 induces expression of SOCS1 and SOCS3 in infected cells, inhibiting the ability of these cells to produce the pro inflammatory, antiviral cytokine IFN-γ (Nowoslawski and Benveniste, 2011). SOCS1 and SOCS3 levels were determined in IFN-γ-treated cells, virus-infected cells, and cells that were both IFN-γ-treated and virus-infected. Results were compared with untreated, uninfected control cells. Flow cytometry data analysis showed a slight decrease in SOCS1 and SOCS3 protein levels in cells treated with IFN-γ for 6 hours compared to control cells. A significant decrease in SOCS1 …

The Construction Of A Plasmid For Detecting The Pathway Of Arginine Metabolism In Human Macrophages: A Real-Time Assessment Of Macrophage Polarity, Benjamin A. Holmes

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Macrophages are "first responders", innate immune system cells which quickly arrive to a site of infection and injury, consuming cell debris and foreign matter and recruiting other immune system cells to the area. While historically they have been thought to react uniformly to all challenges, the discovery of toll-like receptors has shown that macrophages actually work closely with the adaptive immune system in fine-tuning the immune response. Furthermore, it has recently been discovered that macrophages can become polarized to one of two subtypes-M1 or M2.

M1 macrophages are efficient producers of reactive oxygen species, nitrogen intermediates, and inflammatory cytokines. They …

Bacterial Host Interactions In Cystic Fibrosis, Máire Callaghan, Siobhan Mcclean

Articles

Chronic infection is a hallmark of cystic fibrosis (CF) and the main contributor to morbidity. Microbial infection in CF is complex, due to the number of different species that colonise the CF lung. Their colonisation is facilitated by a host response that is impaired or compromised by highly viscous mucous, zones of hypoxia and the lack of the cystic fibrosis transmembrane regulator (CFTR). Successful dominant CF pathogens combine an effective arsenal to establish infection and counter-attack the host response, together with an ability to adapt readily to an unfavourable environment. Hypermutability is common among CF pathogens facilitating adaptation and as …

Coxsackievirus B3 Infection Leads To The Generation Of Cardiac Myosin Heavy Chain-Α-Reactive Cd4 T Cells In A/J Mice, Arunakumar Gangaplara, Chandirasegaran Massilamany, Deborah M. Brown, Gustavo A. Delhon, Asit K. Pattnaik, Nora Chapman, Noel Rose, David J. Steffen, Jay Reddy

Jay Reddy Publications

Enteroviruses like coxsackievirus B3 (CVB3) are common suspects in myocarditis/dilated cardiomyopathy patients. Autoimmunity has been proposed as an underlying mechanism, but direct evidence of its role is lacking. To delineate autoimmune response in CVB3 myocarditis, we used IA^k dextramers for cardiac myosin heavy chain (Myhc)-α 334–352. We have demonstrated that myocarditis-susceptible A/J mice infected with CVB3 generate Myhc-α-reactive CD4 T cells and such a repertoire was absent in naïve mice as measured by proliferative response to Myhc-α 334–352 and IA^k dextramer staining. We also detected Myhc-α 334–352 dextramer⁺ cells in the hearts of CVB3-infected mice. The autoreactive …

Molecular Characterization Of The Host Defense Activity Of The Barrier To Autointegration Factor Against Vaccinia Virus, Nouhou Ibrahim, April Wicklund, Matthew S. Wiebe

Nebraska Center for Virology: Faculty Publications

The barrier to autointegration factor (BAF) is an essential cellular protein with functions in mitotic nuclear reassembly, retroviral preintegration complex stability, and transcriptional regulation. Molecular properties of BAF include the ability to bind double-stranded DNA in a sequence-independent manner, homodimerize, and bind proteins containing a LEM domain. These capabilities allow BAF to compact DNA and assemble higher-order nucleoprotein complexes, the nature of which is poorly understood. Recently, it was revealed that BAF also acts as a potent host defense against poxviral DNA replication in the cytoplasm. Here, we extend these observations by examining the molecular mechanism through which BAF acts …

Activation Of Mmp-9 By Human Lung Epithelial Cells In Response To The Cystic Fibrosis-Associated Pathogen Burkholderia Cenocepacia Reduced Wound Healing In Vitro, Ciara Wright ], Ruth Pilkington, Máire Callaghan, Siobhan Mcclean

Articles

Burkholderia cepacia complex is a group of bacterial pathogens that cause opportunistic infections in cystic fibrosis (CF). The most virulent of these is Burkholderia cenocepacia. Matrix metalloproteinases (MMPs) are upregulated in CF patients. The aim of this work was to examine the role of MMPs in the pathogenesis of B. cepacia complex, which has not been explored to date. Real-time PCR analysis showed that B. cenocepacia infection upregulated MMP-2 and MMP-9 genes in the CF lung cell line CFBE41o- within 1 h, whereas MMP-2, -7, and -9 genes were upregulated in the non-CF lung cell line 16HBE14o-. Conditioned media from …

The Fecal Viral Flora Of Wild Rodents, Tung G. Phan, Beatrix Kapusinszky, Chunlin Wang, Robert K. Rose, Howard L. Lipton, Eric L. Delwart

Biological Sciences Faculty Publications

The frequent interactions of rodents with humans make them a common source of zoonotic infections. To obtain an initial unbiased measure of the viral diversity in the enteric tract of wild rodents we sequenced partially purified, randomly amplified viral RNA and DNA in the feces of 105 wild rodents (mouse, vole, and rat) collected in California and Virginia. We identified in decreasing frequency sequences related to the mammalian viruses families Circoviridae, Picobirnaviridae, Picornaviridae, Astroviridae, Parvoviridae, Papillomaviridae, Adenoviridae, and Coronaviridae. Seventeen small circular DNA genomes containing one or two replicase genes distantly related to the Circoviridae representing several potentially …

Upregulation Of Reactive Oxygen Species During The Retrovirus Life Cycle And Their Roles In A Mutant Of Moloney Murine Leukemia Virus, Ts1-Mediated Neurodegeneration, Soo Jin Kim

Dissertations and Theses (Open Access)

Viral invasion of the central nervous system (CNS) and development of neurological symptoms is a characteristic of many retroviruses. The mechanism by which retrovirus infection causes neurological dysfunction has yet to be fully elucidated. Given the complexity of the retrovirus-mediated neuropathogenesis, studies using small animal models are extremely valuable. Our laboratory has used a mutant moloney murine leukemia retrovirus, ts1-mediated neurodegneration. We hypothesize that astrocytes play an important role in ts1-induced neurodegeneration since they are retroviral reservoirs and supporting cells for neurons. It has been shown that ts1 is able to infect astrocytes in vivo and in …

Novel Mechanisms Of Antigen Processing That Enhance Bcg Vaccine Efficacy, Christopher R. Singh

Dissertations and Theses (Open Access)

Mycobacterium tuberculosis, the causative agent of tuberculosis, is the most lethal single infectious agent afflicting man today causing 2 million deaths per year. The World Health Organization recommends a vaccine as the best option to prevent this disease. The current vaccine, BCG, has a variable efficacy and does not protect adults. It is known that BCG vaccine becomes sequestered in special phagosome compartments of macrophages that do not fuse with lysosomes. Since lysosome fusion is necessary for peptide production and T cell priming leading to protective T_H1 immunity, we hypothesized that vaccine efficacy is reduced and occurs perhaps …

Role Of Porcine Reproductive And Respiratory Syndrome Virus Non-Structural Protein 1 In Viral Replications And Pathogenesis, Lalit Kumar Beura

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Porcine reproductive and respiratory syndrome virus (PRRSV) is an infectious agent of significant concern to the global swine industry. PRRSV infection of pigs is initiated by a long viremia after which the virus enters in an extended persistent phase of 3-4 months that ultimately is resolved by the immune system. The delayed and weak host adaptive response is responsible for the protracted convalescence period. An initial sub-optimal innate immune response is postulated to be the reason behind such meager adaptive immune response. A major focus of the studies undertaken in this dissertation was to identify the viral non-structural proteins (nsps) …

Emerging Dynamics Of Human Campylobacteriosis In Southern Ireland, Susan Bullman, Daniel Corcoran, James O'Leary, Derry O'Hare, Brigid Lucey, Roy D. Sleator

Department of Biological Sciences Publications

Infections with Campylobacter spp. pose a significant health burden worldwide. The significance of Campylobacter jejuni/Campylobacter coli infection is well appreciated but the contribution of non-C. jejuni/C. coli spp. to human gastroenteritis is largely unknown. In this study, we employed a two-tiered molecular study on 7194 patient faecal samples received by the Microbiology Department in Cork University Hospital during 2009. The first step, using EntericBio® (Serosep), a multiplex PCR system, detected Campylobacter to the genus level. The second step, utilizing Campylobacter species-specific PCR identified to the species level. A total of 340 samples were confirmed as Campylobacter genus positive, 329 of …

Archaeoparasitology Of Chaco Canyon, Rachel Paseka

Department of Environmental Studies: Undergraduate Student Theses

Ancient cultures of the Colorado Plateau have been a focus of archaeoparasitology since its inception, and a vast parasitological history is recorded in coprolites preserved in this arid region. The inhabitants of Chaco Canyon dominated Ancestral Puebloan culture between 1050 and 1120 AD and were responsible for the construction of great towns, road systems, and early agriculture. Analysis of the parasites preserved in fecal remains contributes to an increased knowledge of ancient Chacoan health and culture. Nineteen coprolites from four sites in Chaco Canyon were rehydrated and analyzed microscopically for parasite remains. Rhabditiform and filariform nematode larvae were found from …

Role Of Vitamin-D3 And Retinoic Acid In A Human Thp-1 Macrophage Model Of Mycobacterium Tuberculosis Infection, Jaymie L. Estrella

Dissertations and Theses (Open Access)

Mycobacterium tuberculosis (Mtb) replicates within the human macrophages and we investigated the activating effects of retinoic acid (RA) and vitamin D3 (VD) on macrophages in relation to the viability of Mtb. A combination of these vitamins (RAVD) enhanced the receptors on THP-1 macrophage (Mannose receptor and DC-SIGN) that increased mycobacterial uptake but inhibited the
subsequent intracellular growth of Mtb by inducing reactive oxygen species (ROS) and autophagy. RAVD also enhanced antigen presenting and homing receptors in THPs that suggested an activated phenotype for THPs following RAVD treatment. RAVD mediated activation was also associated with a marked phenotypic change in Mtb …

Isolation By Distance Explains Genetic Structure Of Buggy Creek Virus, A Bird-Associated Arbovirus, Abinash Padhi, Amy T. Moore, Mary Bomberger Brown, Jerome E. Foster, Martin Pfeffer, Charles R. Brown

School of Natural Resources: Faculty Publications

Many of the arthropod-borne viruses (arboviruses) show extensive genetic variability and are widely distributed over large geographic areas. Understanding how virus genetic structure varies in space may yield insight into how these pathogens are adapted to and dispersed by different hosts or vectors, the relative importance of mutation, drift, or selection in generating genetic variability, and where and when epidemics or epizootics are most likely to occur. However, because most arboviruses tend to be sampled opportunistically and often cannot be isolated in large numbers at a given locale, surprisingly little is known about their spatial genetic structure on the local …

Influenza Nucleoprotein Interactions As Potential Antiviral Targets, Abel Sanchez

Theses Digitization Project

The purpose of this research was to investigate the Nucleoprotein (NP) RNA dependent RNA polymerase (RdRP) interaction by identifying and characterizing a NP mutant which maintains RNA binding but is unable to interact with the viral polymerase complex. Influenza poses a threat to human health and is a burden on the global economy. Influenza is caused by influenza virus which contains a segmented RNA genome, resulting in rapid evolution and emergence of new influenza subtypes which lead to pandemics.

Utilization Of A Custom-Designed Microbiota Array To Determine The Distal Gut Microbiota Of Healthy Human Adults, Richard Thomas Agans

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The human microbiota is an essential component of human health and disease. It is involved in metabolism of dietary components and is at the forefront of the intestinal immune response. Classical techniques applied to study intestinal microbiota illustrated higher presences of aerobic and facultative-anaerobic bacteria, however; the levels of obligate anaerobes had been underrepresented. Modern technologies based on DNA and RNA analysis have circumvented previous challenges allowing researchers to gain more extensive insight into the complex intestinal environment. In this work, a recently developed Microbiota Array was used to assess intestinal microbiota of 10 healthy adults (age 22-61, ave. 34.3yrs). …

Identification Of The Role Of The Sal Locus In Streptococcus Pyogenes Virulence During Host-Pathogen Interactions, Phanramphoei Namprachan-Frantz

Wayne State University Dissertations

The pathogenesis of Streptococcus pyogenes is due to its ability to overcome and adapt to the harsh environment created by the host immune response. The focus of this project was the SalKR two-component regulatory system, which facilitates bacterial adaptation by responding to environmental signals during host pathogen-interactions. The first goal of this project was to determine a role in virulence for the SalKR regulatory system. The complete deletion of the salKR genes in the wild type S. pyogenes strain HSC5 produced a highly attenuated mutant in a Zebrafish infection model. The ΔsalKR mutant appeared to lose the ability to survive …