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Generation And Application Of Antigen-Specific Induced Regulatory T Cells In Allogeneic Bone Marrow Transplantation, Kenrick Semple

USF Tampa Graduate Theses and Dissertations

CD28 co-stimulation is required for the generation of naturally occurring regulatory T cells (nTregs) in the thymus through Lck-signaling. However, high level of CD28 suppresses the generation of induced Tregs (iTregs) from naïve CD4 T cells, although underlying mechanism(s) has not been defined. Here we investigated the role of CD28-mediated signaling pathways in the suppression of Treg generation. We used a series of transgenic (Tg) mice on CD28-deficient background that bears WT CD28 or mutated CD28 in its cytosolic tail incapable of binding to Lck, PI3K or Itk. Regardless of exogenous IL-2, strong CD28 costimulation suppressed iTreg generation through Lck …

Novel Roles For The Transcriptional Repressor Prdm1 In Human Natural Killer Cells And Identification Of An Inhibitor Of Its Interacting Methyltransferase G9a, Matthew Adams Smith

USF Tampa Graduate Theses and Dissertations

The studies presented within this dissertation provide the first description of PRDM1 (also known as Blimp-1 or PRDI-BF1) function in natural killer cells. NK cells are major effectors of the innate immune response via antigen-independent cytotoxicity and link to the adaptive immune response through cytokine release. Molecular mechanisms mediating NK activation are relatively well-studied; however, much less is known about the mechanisms that restrain activation.

In the first study, the transcriptional repressor PRDM1 is shown to be a critical negative regulator of NK function. Microarray analysis was used to characterize transcriptional changes associated with cytokine-mediated activation. PRDM1 is expressed at …

Development Of A Non-Invasive Electrode For Intradermal Electrically Mediated Dna Vaccination, Amy Lynn Donate

USF Tampa Graduate Theses and Dissertations

Current progress in the development of vaccines has decreased the incidence of fatal and non-fatal infections and increased longevity. However, new technologies need to be developed to combat an emerging generation of infectious diseases. DNA vaccination has been demonstrated to have great potential for use against a wide variety of diseases. Alone, this vaccine technology does not generate a significant immune response for vaccination, but combined with delivery by electroporation (EP), can enhance plasmid expression and immunity against the expressed antigen. Most EP systems, while effective, can be invasive and painful making them less desirable for use in vaccination. Our …

Ctla-4 Transcriptional Activation: Regulation Of Induced Expression, Heather Gibson

Wayne State University Dissertations

Cytotoxic T-lymphocyte antigen 4 (CTLA-4) is a T cell surface protein that is homologous to CD28 and binds to the B7 family of ligands. Unlike CD28, CTLA-4 interaction transmits a negative signal in T cells, leading to suppression of proliferation. CTLA-4 is constitutively expressed on regulatory T cells (Tregs) but is also inducible in effector T cells. The mechanisms driving transcriptional regulation of CTLA-4 are poorly understood. Our previous work identified a bona fide NFAT1 binding site in the proximal promoter for effector T cells. In addition, we found histone acetylation occurred after stimulation.

As a result of its role …

Identification And Characterization Of Equine Rotaviruses In Southern Ireland, Crystal Cronin

Theses

Group A Rotaviruses(GARVs) are enteric pathogens which induce acute gastroenteritis in infants, neonates and the young of many animal species. During a 4 year study (2005-2009), 64 fecal samples from symptomatic foals, from several counties, in the south of Ireland, were molecularly characterized to determine the VP& (G type), VP4 (P type), and NSP4 antigenic specificities. Globally, G3 [P12] and G14[P12] are recognized as the predominant equine GARV strains. Twenty-two samples (34.38%) were found to contain G3 viruses, while 21 samples (32.81%) contained G14 viruses. In this study, there was a distinct increase in the incidence of mixed infections circulating …

Hsv-1 Infection In Keratinocyte Cell Lines Treated With Mitotic Inhibitors, Asma A. Abbas

Browse all Theses and Dissertations

The hypothesis for this research was: Herpes simplex virus type 1 (HSV-1) infection of murine keratinocyte cell lines (HEL-30 and PAM-212) treated with mitotic inhibitors leads to silencing of virus replication. At 0.1 (Multiplicity of infection) MOI, PAM-212 keratinocytes showed some cell lysis and viral plaques were seen in HEL-30 keratinocyte cultures. PAM-212 keratinocytes, infected at 0.01 MOI, were more susceptible to the lytic effect of HSV-1 than were HEL-30 cells. In this study, 5-fluoro-2'-deoxyuridine (FUDR) treatment of both keratinocyte cell lines permitted an increase survival of HSV-1- infected keratinocytes especially for PAM-212 cell line which was further examined for …

Distribution Of Cellular Interferon Beta (Ifn-Β) In Murine Fibroblast Cell Lines Upon Infection Of Hsv-1, Rachael E. Curtis

Browse all Theses and Dissertations

The hypothesis for this study was: IFN-β expression differs between murine fibroblasts (A2R1 and L929) of different mouse strains upon infection of HSV-1. Fluorescent microscopy was used to examine localization of IFN-β in the different murine fibroblast and keratinocyte cell lines after 6 hours of infection with herpes simplex virus type-1 (HSV-1). Because keratinocytes cell lines HEL-30 and PAM-212 grew in clusters, staining patterns in individual cells could not be determined. A notable difference in localization of IFN-β immune staining was seen between the two cell lines when infected with HSV-1. The A2R1 cell line showed perinuclear (PN) localization, while …

Targeted Knockdown Of Amp-Activated Protein Kinase Alpha 1 And Alpha 2 Catalytic Subunits, Larissa J. Tangeman

Browse all Theses and Dissertations

AMP-activated protein kinase (AMPK) regulates cellular metabolism and promotes ATP production when energy is depleted. Evidence suggests that AMPK may be involved in oxygen sensing by carotid body cells, which are responsible for regulating the breathing rate to maintain proper blood oxygen levels. There are two isoforms of the catalytic alpha subunit, AMPKalpha1 and AMPKalpha2, which could be involved in oxygen sensing. Here, the production of a short hairpin RNA (shRNA) targeting both catalytic isoforms of AMPK in human, mouse, and rat is described. The shRNA causes significant knockdown of both isoforms of AMPK alpha in mouse and human cells …

Strategies For Long-Term Renal Allograft Survival In Iga Nephropathy Patients, Erin E. Lintz

Browse all Theses and Dissertations

IgA nephropathy is one of the most common kidney diseases worldwide. Most IgA nephropathy patients will undergo a kidney transplant as a treatment. Treatment currently includes the use of immunosuppressants which are necessary to prevent graft rejection but present harmful side-effects when taken long-term. This review focuses on strategies which have the ability to promote tolerance of both donor and recipient cells within a transplant recipient. Such strategies allow development of mixed chimerism and thereby alleviate the need for long-term immunosuppressant usage in allograft recipients. These strategies could be applied to IgA nephropathy patients to allow kidney allograft acceptance without …

Amino Acid Substitutions Created In Reverse Transcriptase And Their Influence On Hiv-1 Mutation Frequencies, Amani Saud Alhejely

Browse all Theses and Dissertations

As research has shown, a high mutation rate occurs during the reverse transcription RT process because HIV reverse transcriptase fails to correct erroneously incorporated nucleotides during the reverse transcription process. Based on various research articles, a series of amino acid substitutions created in RT were collected to find out their influence on virus mutation frequencies. Two tables were used; one for amino acid substitutions created in RT that increased the HIV-1 mutation rate and another for amino acid substitutions created in RT that decreased the HIV-1 mutation rate. The tables show which amino acid substitutions created in RT decreased or …

Developing A Quantitative Pcr Assay For Detecting Viral Vector Shedding From Animals, Swathee Chinnasamy

Browse all Theses and Dissertations

Replication deficient viruses have been used widely for replacing, repairing, and deleting target genes. These recombinant viruses are tested on research animals or patients in clinical trials. Although viral vectors distribute in the body, they are also disseminated into the environment through secretion and excretion processes. By studying the extent of shedding, a proper risk assessment can be performed and appropriate biocontainment can be achieved. Adenoviral and lentiviral vectors were produced from commercially available kits. The transgene present in both vector systems was the lacZ reporter gene encoding for β-galactosidase. Primers and probes were designed for the encapsidation region of …

Some Laelapine Mites (Acari: Laelapidae) Ectoparasitic On Small Mammals In The Galapagos Islands, Including A New Species Of Gigantolaelaps From Aegialomys Galapagoensis, Donald D. Gettinger, Fernanda Martins-Hatano, Scott Lyell Gardner

Scott L. Gardner Publications

A collection of laelapine mites from small mammals in the Galapagos Islands are identified and their host distributions reviewed. Two species of native rodents, Aegialomys galapagoensis and Nesoryzomys narboroughii, were infested only with species typical of Neotropical oryzomyine rodents; Rattus rattus was infested with Laelaps nuttalli, a host-specific ectoparasite to Old World Rattus. A synopsis of Gigantolaelaps Fonseca is provided and we describe a new laelapine mite, Gigantolaelaps aegialomys n. sp., from the pelage of the rodent A. galapagoensis on Santa Fe Island. The new species has strong morphological affinities with a subgroup of Gigantolaelaps associated …

Carbenoxolone Blocks The Light-Evoked Rise In Intracellular Calcium In Isolated Melanopsin Ganglion Cell Photoreceptors, Jayne R. Bramley, Erin M. Wiles, Patricia J. Sollars, Gary E. Pickard

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: Retinal ganglion cells expressing the photopigment melanopsin are intrinsically photosensitive (ipRGCs). These ganglion cell photoreceptors send axons to several central targets involved in a variety of functions. Within the retina ipRGCs provide excitatory drive to dopaminergic amacrine cells via glutamatergic signals and ipRGCs are coupled to widefield GABAergic amacrine cells via gap junctions. However, the extent to which ipRGCs are coupled to other retinal neurons in the ganglion cell layer via gap junctions is unclear. Carbenoxolone, a widely employed gap junction inhibitor, greatly reduces the number of retinal neurons exhibiting non-rod, non-cone mediated light-evoked Ca²⁺ signals suggesting extensive …

Intrinsically Photosensitive Retinal Ganglion Cells, Gary E. Pickard, Patricia J. Sollars

School of Veterinary and Biomedical Sciences: Faculty Publications

Intrinsically photosensitive retinal ganglion cells (ipRGCs) respond to light in the absence of all rod and cone photoreceptor input. The existence of these ganglion cell photoreceptors, although predicted from observations scattered over many decades, was not established until it was shown that a novel photopigment, melanopsin, was expressed in retinal ganglion cells of rodents and primates. Phototransduction in mammalian ipRGCs more closely resembles that of invertebrate than vertebrate photoreceptors and appears to be mediated by transient receptor potential channels. In the retina, ipRGCs provide excitatory drive to dopaminergic amacrine cells and ipRGCs are coupled to GABAergic amacrine cells via gap …

Long-Term Clinicopathological Characteristics Of Alpacas Naturally Infected With Bovine Viral Diarrhea Virus Type Ib, D. Bedenice, Edward J. Dubovi, Clayton Kelling, Jamie N Henningson, Christina L. Topliff, N. Parry

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: Substantial bovine viral diarrhea virus (BVDV)-related production losses in North American alpaca herds have been associated with BVDV type Ib infection.

Objectives: To classify and differentiate the long-term clinicopathological characteristics of BVDV type Ib infection of al- paca crias, after natural virus exposure. We hypothesized that persistently infected (PI) alpacas specifically demonstrate growth retardation, clinicopathological evidence of opportunistic infections, and early mortality.

Animals: Thirty-five crias naturally exposed to BVDV (18 acute, 3 chronic, 14 PIs), and 19 healthy cohort controls of 5 northeastern alpaca farms were prospectively evaluated over 2 years (September 2005–September 2008).

Methods: Observational cohort-control study.

Results: …

Consensus-Based Reporting Standards For Diagnostic Test Accuracy Studies For Paratuberculosis In Ruminants, Ian A. Gardner, Søren S. Nielsen, Richard J. Wittington, Michael T. Collins, Douwe Bakker, Beth Harris, Srinand Sreevatsan, Jason E. Lombard, Raymond Sweeney, David R. Smith, Jerrie Gavalchin, Shigetoshi Eda

School of Veterinary and Biomedical Sciences: Faculty Publications

The Standards for Reporting of Diagnostic Accuracy (STARD) statement was developed to encourage complete and transparent reporting of key elements of test accuracy studies in human medicine. The statement was motivated by widespread evidence of bias in test accuracy studies and the finding that incomplete or absent reporting of items in the STARD checklist was associated with overly optimistic estimates of test performance characteristics. Although STARD principles apply broadly, specific guidelines do not exist to account for unique considerations in livestock studies such as herd tests, potential use of experimental challenge studies, a more diverse group of testing purposes and …

Erection Of Ibirhynchus Gen. Nov. (Acanthocephala: Polymorphidae), Based On Molecular And Morphological Data, Martin Garcia-Varela, Gerardo Pérez-Ponce De León, Francisco J. Aznar, Steven A. Nadler

Harold W. Manter Laboratory of Parasitology: Faculty Publications

The genus Southwellina is composed of 3 described species, i.e., S. hispida (the type species), S. dimorpha, and S. macracanthus. All 3 are endoparasites of fish-eating birds that have worldwide distributions. Morphologically, the genus is characterized by possessing a short and compact trunk, 2 fields of spines in the anterior region of the trunk (in at least 1 sex), a short cylindrical proboscis (sometimes with a swollen region armed with numerous longitudinal rows of hooks), a double-walled proboscis receptacle, and 4 tubular cement glands in males. In the current study, specimens identified as S. dimorpha were collected from …

Helminths Of Squirrels (Sciuridae) From Mongolia, David S. Tinnin, Sumiya Ganzorig, Scott Lyell Gardner

Harold W. Manter Laboratory of Parasitology: Faculty Publications

Two species of ground squirrels, Urocitellus undulatus and Spermophilus alashanicus, were collected and examined for parasites from two localities in Mongolia in 1999. A total of 24 individuals of U. undulatus were examined resulting in a total of six taxa of helminths found, including: Ascaris tarbagan, Physaloptera massino, Streptopharagus kutassi, Anoplocephaloides transversaria, Hymenolepis suslica, and Moniliformis sp. This represents four new species records and an additional host record for the country. Two individuals of S. alashanicus were examined and found to be uninfected.

Description Of Scottnema Lindsayae Timm, 1971 (Rhabditida: Cephalobidae) From Taylor Valley, Antarctica And Its Phylogenetic Relationship, Sven Boström, Oleksandr Holovachov, Steven A. Nadler

Harold W. Manter Laboratory of Parasitology: Faculty Publications

The endemic Antarctic nematode Scottnema lindsayae is described from specimens collected in Taylor Valley, McMurdo Dry Valleys, Victoria Land. The recently collected material is compared with the original description and other subsequent descriptions of the species. A more complete scanning electron microscopy (SEM) study of the species is presented. The phylogenetic position of S. lindsayae is inferred using a secondary structure-based alignment of a partial sequence of nuclear Large Subunit (LSU) ribosomal DNA. Phylogenetic trees were inferred using base-paired substitution models implemented in PHASE 2 software and Bayesian inference, and show S. lindsayae as the sister group to Stegelletina taxa.

Where Have All The Plant Foods Gone? The Search For Refined Dietary Reconstruction From Chinchorro Mummies, Karl J. Reinhard, Sara Le-Roy-Toren, Bernardo Arriaza

Karl Reinhard Publications

The Chinchorro cemeteries of northern Chile are the first graveyards of the Andes (Arriaza 1995, Schiappacasse & Niemeyer 1984). The excavation and analysis of Chinchorro mummies reveals the 'true character of the people'. The burials themselves are optimal for preservation of corpses, burial furniture, and aspects of ceremonies associated with the dead. The analysis of corporeal remains gradually clarifies the details of daily activities and status. Therefore, we feel that a holistic approach to mummy studies provides enormously fruitful paths to understanding the character of the ancient Chinchorro society.

Paleoepidemiology Of Intestinal Parasites And Lice In Pre-Columbian South America, Adauto Araujo, Karl Reinhard, Daniela Leles, Luciana Sianto, Alena M. Iñiguez, Martin Fugassa, Bernardo Arriaza, Nancy Orellana, Luis Fernando Ferreira

Karl Reinhard Publications

Some human parasites originated in prehominid ancestors in Africa. Nematode species, such as Enterobius vermicularis (pinworm), hookworms and Trichuris trichiura are shared by humans and other close phylogenetic primates (Pan and Gorilla), showing that they infected a common ancestor to this group. When humans migrated from Africa to other continents they carried these parasites wherever climate conditions allowed parasite transmission from host to host. Other parasites, however, were acquired throughout human biological and social evolutive history when new territories were occupied. Paleoparasitology data is a valuable source to recover emergence and disappearance of parasite infections through analysis of …

Alternations In The Splenic Marginal Zone With Age, Shirin Z. Birjandi

Dissertations

Splenic marginal zones are architecturally organized to generate a rapid response against blood- borne antigens entering the spleen. The marginal zone is a distinct anatomical micro-environment whose main components include the marginal zone macrophages, the marginal zone B cells, the marginal zone sinus, and the metallophilic macrophages. Marginal zone macrophages, in partnership with marginal zone B cells, are particularly important in host defense against T-independent pathogens and are crucial for the prevention of diseases such as Streptococcus pneumonia. It has been widely reported that with the advancement of age there is a higher rate of mortality as a result of …

Role For Histone Deacetylases In Glucocorticoid Receptor Mediated Transpression Of Natural Killer Cell Activity, Kristin Bush

Master's Theses

During periods of psychosocial distress glucocorticoids (GCs) are known to reduce the lytic activity of natural killer cells (NKCA). Glucocorticoid treatment also reduces acetylation of histone residues; however, the glucocorticoid receptor (GR) lacks deacetylase activity. GR is known to interact with histone deacetylases (HDACs) and with corepressors that mediate gene transrepression. In this investigation, GC induced histone deacetylation was demonstrated to be due to GR recruitment of HDAC1 and the corepressor complex SMRT. These data show that reduced acetylation of immune functional genes associated with NKCA is likely due to histone deacetylation by HDAC1 and transrepression of those genes by …

Estimating The Reproductive Numbers For The 2008-2009 Cholera Outbreaks In Zimbabwe, Zindoga Mukandavire, Shu Liao, Jin Wang, Holly Gaff, David L. Smith, J. Glenn Morris Jr.

Biological Sciences Faculty Publications

Cholera remains an important global cause of morbidity and mortality, capable of causing periodic epidemic disease. Beginning in August 2008, a major cholera epidemic occurred in Zimbabwe, with 98,585 reported cases and 4,287 deaths. The dynamics of such outbreaks, particularly in nonestuarine regions, are not well understood. We explored the utility of mathematical models in understanding transmission dynamics of cholera and in assessing the magnitude of interventions necessary to control epidemic disease. Weekly data on reported cholera cases were obtained from the Zimbabwe Ministry of Health and Child Welfare (MoHCW) for the period from November 13, 2008 to July 31, …

Differential Innate Immune Responses Correlate With The Contrasting Pathogenicity Of The Equine H7n7 Influenza Virus Demonstrated In Horses And Balb/C Mice, Liang Zhang

University of Kentucky Doctoral Dissertations

Equine influenza virus causes a mild, self-limiting upper respiratory disease in its natural host. In stark contrast, equine influenza viruses of the H7N7 subtype produce lethal infection in BALB/c mice. This dissertation explored the mechanism underlying the differential pathogenicity of the equine H7N7 influenza virus observed in horses and BALB/c mice. Initially, a comparative study of the pathogenesis was conducted in BALB/c mice inoculated intranasally with a representative isolate of either H7N7 or H3N8 subtype equine influenza virus. All H3N8 virus-infected mice survived the infection whereas 100% mortality was documented for the mice receiving the H7N7 virus by day 8 …

An Investigation Of The Subtype Diversity Of Clinical Isolates Of Irish Clostridium Difficile Ribotypes 027 And 078 By Repetitive-Extragenic Palindromic Pcr, Denise Drudy, K. Solomon, S. Murray, L. Scott, S. Mcdermott,, A. Martin, C. O’Donoghue, M. Skally, K. Burns, L. Fenelon, F. Fitzpatrick,, L. Kyne, S. Fanning

Articles

A repetitive-extragenic palindromic PCR (rep-PCR) subtyping method (DiversiLab) in conjunction with ribotyping, toxinotyping and antimicrobial-susceptibility testing was used to detect subtypes within Clostridium difficile ribotypes 027 and 078. Clinical isolates of ribotypes 027 (toxinotype III) (n530) and 078 (toxinotype V) (n523) were provided by health-care facilities across the Republic of Ireland over 2 months in 2006 and 1 month in 2009. Ribotype 027 isolates were significantly more related to each other (9 different subtype profiles) when compared to ribotype 078 isolates (14 different profiles) (P50.001; cut-off .90 % similarity). Almost half of ribotype 078 isolates (45.5 %) showed no relationship …

The Effects Of Myd88 Inhibition On Bovine Lung Endothelial And Epithelial Cell Cytokine Expression, Cassandra Kay Hayne

Honors Program Theses

Bovine respiratory disease complex (BRDC) has major economical implications on the beef industry due to the severe health problems it produces in calves. Although several organisms are associated with it, Mannheimia haemolytica induces the severest amount of damage to the lungs. It has previously been demonstrated that lipopolysaccharide (LPS) produced by this bacterium causes significant permeability increases and cell apoptosis in bovine lung endothelial cells (BPMEC) but not bovine lung epithelial cells (TBPE). The purpose was to determine whether the main signaling pathway for cells that respond to LPS signaling, the Myeloid Differentiation 88 Pathway (MYD88), is present and functional …

Tca Cycle Inactivation In Staphylococcus Aureus Alters Nitric Oxide Production In Raw 264.7 Cells, Chandirasegaran Massilamany, Arunakumar Gangaplara, Donald J. Gardner, James M. Musser, David J. Steffen, Greg A. Somerville, Jay Reddy

Jay Reddy Publications

Inactivation of the Staphylococcus aureus tricarboxylic acid (TCA) cycle delays the resolution of cutaneous ulcers in a mouse soft tissue infection model. In this study, it was observed that cutaneous lesions in mice infected with wild-type or isogenic aconitase mutant S. aureus strains contained comparable inflammatory infiltrates, suggesting the delayed resolution was independent of the recruitment of immune cells. These observations led us to hypothesize that staphylococcal metabolism can modulate the host immune response. Using an in vitro model system involving RAW 264.7 cells, the authors observed that cells cultured with S. aureus aconitase mutant strains produced significantly lower amounts …

Rnai Screening Reveals Requirement For Host Cell Secretory Pathway In Infection By Diverse Families Of Negative-Strand Rna Viruses, Debasis Panda, Anshuman Das, Phat X. Dinh, Sakthivel Subramaniam, Debasis Nayak, Nicholas J. Barrows, James L. Pearson, Jesse Thompson, David L. Kelly, Istvan Ladunga, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

Negative-strand (NS) RNA viruses comprise many pathogens that cause serious diseases in humans and animals. Despite their clinical importance, little is known about the host factors required for their infection. Using vesicular stomatitis virus (VSV), a prototypic NS RNA virus in the family Rhabdoviridae, we conducted a human genomewide siRNA screen and identified 72 host genes required for viral infection. Many of these identified genes were also required for infection by two other NS RNA viruses, the lymphocytic choriomeningitis virus of the Arenaviridae family and human parainfluenza virus type 3 of the Paramyxoviridae family. Genes affecting different stages of …

Y-Like Retinal Ganglion Cells Innervate The Dorsal Raphe Nucleus In The Mongolian Gerbil (Meriones Unguiculatus), Liju Luan, Chaoran Ren, Benson Wui-Man Lau, Jian Yang, Gary E. Pickard, Kwok-Fai So, Mingliang Pu

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: The dorsal raphe nucleus (DRN) of the mesencephalon is a complex multi-functional and multi-transmitter nucleus involved in a wide range of behavioral and physiological processes. The DRN receives a direct input from the retina. However little is known regarding the type of retinal ganglion cell (RGC) that innervates the DRN. We examined morphological characteristics and physiological properties of these DRN projecting ganglion cells.

Methodology/Principal Findings: The Mongolian gerbils are highly visual rodents with a diurnal/crepuscular activity rhythm. It has been widely used as experimental animals of various studies including seasonal affective disorders and depression. Young adult gerbils were used …