Immunology and Infectious Disease Commons^™

Characterization Of A Putative Cephalotoxin From The Hemocytes Of Euprymna Scolopes, Jessie E. Scott

University Scholar Projects

Many organisms have beneficial microbial symbionts, and complex interactions between the immune system and the bacteria must occur to allow these symbioses to persist. The host immune system must recognize and preferentially tolerate the beneficial microorganisms, while simultaneously eliminating harmful pathogens. The Hawaiian bobtail squid, Euprymna scolopes, is a model organism for host-microbe interactions because of the unique light organ symbiosis. Hemocytes, the squid’s primary immune cell, infiltrate and patrol the light organ and continuously interact with the symbiont Vibrio fischeri. Using this model, several innate immunity genes have been shown to be differentially expressed in the hemocytes …

Regulation Of Macrophage Inflammatory Signaling Pathways By Amp-Activated Protein Kinase., Yanfang Zhu

Electronic Theses and Dissertations

AMP-activated protein kinase, AMPK, is a conserved serine/threonine kinase with a critical function in the regulation of metabolic pathways in eukaryotic cells. Recently, AMPK has been shown to play an additional role as a regulator of inflammatory activity in leukocytes. Treatment of macrophages with chemical AMPK activators, or forced expression of a constitutively active form of AMPK, results in polarization to an antiinflammatory phenotype. Additionally, we reported previously that stimulation of macrophages with antiinflammatory cytokines such as IL-10, IL-4 and TGF-ß results in rapid activation of AMPK, suggesting that AMPK contributes to the suppressive function of these cytokines. In the …

Sa-4-1bbl As A Platform To Develop Adjuvant Systems For Prophylactic And Therapeutic Vaccines., Gunes Dinc 1984-

Electronic Theses and Dissertations

Vaccines against infectious diseases are one of the most critical accomplishments in modern medicine. Despite significant progress in vaccinology, there is still a dire need for developing vaccines against various acute and chronic infections and cancer. In general, vaccines are categorized as prophylactic, given to healthy individuals to prevent disease, and therapeutic, administered to people who already have disease. As such, the nature, quality, and quantity of immune responses required for the efficacy of these two types vaccines are different. Prophylactic vaccines against infectious diseases primarily rely on the generation of neutralizing high titers of antibody for their efficacy. These …

Reciprocity In Microbiome And Immune System Interactions And Its Implications In Disease And Health., Enayat Nikoopour, Bhagirath Singh

Microbiology & Immunology Publications

Adaptation of the whole microbial normal flora residing in a host to its natural habitat over an evolutionary period has resulted in peaceful coexistence with mutual benefits for both microbiota and host in steady state. This symbiotic relationship between host and microbiota has a significant impact on shaping the immune response in the host to achieve an immune tolerance to microbiota but retaining the ability to respond to invading pathogens. Perturbation of this balance by manipulation of microbial communities in the host can lead to immune dysregulation and susceptibility to diseases. By studying the host in the absence of microbiota …

Deletion Of The Human Cytomegalovirus Us17 Gene Increases The Ratio Of Genomes Per Infectious Unit And Alters Regulation Of Immune And Endoplasmic Reticulum Stress Response Genes At Early And Late Times After Infection, Stephen James Gurczynski, Subhendu Das, Philip E. Pellett

Immunology and Microbiology Faculty Research Publications

Human cytomegalovirus (HCMV) employs numerous strategies to combat, subvert, or co-opt host immunity. One evolutionary strategy for this involves capture of a host gene and then its successive duplication and divergence, forming a family of genes, many of which have immunomodulatory activities. The HCMV US12 family consists of 10 tandemly arranged sequence-related genes in the unique short (US) region of the HCMV genome (US12 to US21). Each gene encodes a protein possessing seven predicted transmembrane domains, patches of sequence similarity with cellular G-protein-coupled receptors, and the Bax inhibitor 1 family of antiapoptotic proteins. We show that one member, US17, plays …

Role Of Peripheral Immune Response In Microglia Activation And Regulation Of Brain Chemokine And Proinflammatory Cytokine Responses Induced During Vsv Encephalitis, Christina D. Steel, Kimberly Breving, Susan Tavakoli, Woong-Ki Kim, Larry D. Sanford, Richard P. Ciavarra

Biological Sciences Faculty Publications

We report herein that neuroinvasion by vesicular stomatitis virus (VSV) activates microglia and induces a peripheral dendritic cell (DC)-dependent inflammatory response in the central nervous system (CNS). VSV neuroinvasion rapidly induces multiple brain chemokine and proinflammatory cytokine mRNAs that display bimodal kinetics. Peripheral DC ablation or T cell depletion suppresses the second wave of this response demonstrating that infiltrating T cells are primarily responsible for the bimodal characteristics of this response. The robust infiltrate associated with VSV encephalitis likely depends on sustained production of brain CCL19 and CCR7 expression on infiltrating inflammatory cells. © 2013 Elsevier B.V. All rights reserved.

Candida Albicans Als5p Amyloid In Host-Microbe Interactions: A Ceanorhabditis Elegans Study, Michael Bois

Dissertations, Theses, and Capstone Projects

Candida albicans, a dimorphic fungus and an opportunistic pathogen, possesses a myriad of adherence factors including members of the agglutinin-like sequence (Als) family of mannoproteins. The adhesin Als5p mediates adhesion to many substrates, and is upregulated during commensal interactions, but is downregulated during active C. albicans infections[1]. An amyloid forming core sequence at residues 325-331 has been shown to be important for Als5p function, because a single amino acid substitution at position 326 (V326N) greatly reduces Als5p-mediated adherence[2]. We evaluated the role of Als5p in host-microbe interactions, using Caenorhabditis elegans as a host model and feeding them Saccharomyces cerevisiae expressing …

Phages Of Non-Diary Lactococci: Isolation And Characterization Of Phi L47, A Phage Infecting The Grass Isolate Lactococcus Lactis Ssp Cremoris Dpc6860, Daniel Cavanagh, Caitríona M. Guinane, Horst Neve, Aidan Coffey, R. Paul Ross, Gerald F. Fitzgerald, Olivia Mcauliffe

Department of Biological Sciences Publications

Lactococci isolated from non-dairy sources have been found to possess enhanced metabolic activity when compared to dairy strains. These capabilities may be harnessed through the use of these strains as starter or adjunct cultures to produce more diverse flavor profiles in cheese and other dairy products. To understand the interactions between these organisms and the phages that infect them, a number of phages were isolated against lactococcal strains of non-dairy origin. One such phage, ΦL47, was isolated from a sewage sample using the grass isolate L. lactis ssp. cremoris DPC6860 as a host. Visualization of phage virions by transmission electron …

Immunologic Special Forces: Anti-Pathogen Cytotoxic T-Lymphocyte Immunotherapy Following Hematopoietic Stem Cell Transplantation, Michael Keller, Catherine M. Bollard

Pediatrics Faculty Publications

Anti-pathogen adoptive T-cell immunotherapy has been proven to be highly effective in preventing or controlling viral infections following hematopoietic stem cell transplantation. Recent advances in manufacturing protocols allow an increased number of targeted pathogens, eliminate the need for viral transduction, broaden the potential donor pool to include pathogen-naïve sources, and reduce the time requirement for production. Early studies suggest that anti-fungal immunotherapy may also have clinical benefit. Future advances include further broadening of the pathogens that can be targeted and development of T-cells with resistance to pharmacologic immunosuppression.

Construction Of A Live-Attenuated Hiv-1 Vaccine Through Genetic Code Expansion, Nanxi Wang, Yue Li, Wei Niu, Ming Sun, Ronald Cerny, Qingsheng Li, Jiantao Guo

Qingsheng Li Publications

A safe and effective vaccine against human immunodeficiency virus type 1 (HIV-1) is urgently needed to combat the worldwide AIDS pandemic, but still remains elusive. The fact that uncontrolled replication of an attenuated vaccine can lead to regaining of its virulence creates safety concerns precluding many vaccines from clinical application. We introduce a novel approach to control HIV-1 replication, which entails the manipulation of essential HIV-1 protein biosynthesis through unnatural amino acid (UAA*)-mediated suppression of genome-encoded blank codon. We successfully demonstrate that HIV-1 replication can be precisely turned on and off in vitro.

Includes supporting information.

Glut 4 And Insulin Resistance, Salmin Ali

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Five types of glucose transporters stimulate the transport of glucose from the bloodstream into cells such as muscle cells and adipocytes in response to insulin signaling or exercise. Skeletal muscle and adipose tissue are the main sites for glucose uptake and dispose in insulin action dependent pathway. In non-insulin stimulated condition, the GLUT4 storage vesicles stay away from the plasma membrane, so there is no GLUT4 translocation or GLUT4 mediated glucose uptake. However, in insulin stimulated state, insulin initiates insulin signaling cascade which results in rapid GLUT4 translocation and increased glucose uptake at the cell surface. Failure of the translocation …

Differentiation Of U-937 Monocytes To Macrophage-Like Cells Polarized Into M1 Or M2 Phenotypes According To Their Specific Environment: A Study Of Morphology, Cell Viability, And Cd Markers Of An In Vitro Model Of Human Macrophages, Fatma Husien S. Abdulhadi

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In this study, the human leukemic monocyte lymphoma cell line U-937 was used as an in vitro model for monitoring monocyte/macrophage differentiation. Phorbol 12- myristate (13) (PMA) was used to activate U-937 cells into macrophage-like cells (M0). After 24 hours of PMA treatment, non- adherent U-937 cells became tightly adherent to the culture plates forming M0 cells. M0 cells were then polarized into the M1 macrophage phenotype by treatment with LPS and IFN-γ for another 24 hours. Each of the cytokines IL-4, IL-13, or IL-10 was applied separately to three M0 cultures for 24 hours to induce the M2 macrophage …

Role Of Viral And Host Factors In Influenza Virus Mediated Inhibition Of Interleukin-23, Ashish Tiwari

Theses and Dissertations--Veterinary Science

Influenza virus is one of the major respiratory pathogens of humans as well as animals, including equines. There is an increasing evidence that bacterial infections are the most common cause of the death during influenza. In horses also, secondary bacterial pneumonia can lead to death, and surviving horses may take up to six months for the complete recovery resulting in heavy economic loss to the equine industry. Interleukin (IL)-23 mediated innate immune response has been shown to protect the host from various respiratory bacterial infections. However, studies to investigate the role of host and viral factors in the regulation of …

Socs1/Socs3 Expression And Virus Replication Of Denv2 And Hsv-1 In Cytokine-Polarized Subsets Of Raw 264.7 Macrophages, Nagarjuna Reddy Cheemarla

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Cytokine activation of macrophages leads to "macrophage polarizatio" The M1 subset is polarized by exposure to lipopolysaccharide (LPS) and interferon-gamma (IFN-γ) and M2 polarized cells develop after exposure to interleukin-4 (IL-4) or IL-13. In this study, the RAW 264.7 murine macrophage cell line was used to study the effect herpes simplex virus-Type I (HSV-1) and Dengue virus serotype-2 (DENV2) infections have on cultures of unpolarized (M0) or polarized subsets. The macrophage subsets were characterized using cluster of differentiation markers CD14 and CD86. Uninfected M1 macrophages showed up regulation in expression of the co-stimulatory factor B7.2 (CD86) compared to expression by …

Development Of A Co-Culture System To Mimic The Transfection Of Hsv-1 From Keratinocytes To Neuronal Cells, David A. Dixon

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To more closely mimic the in vivo progression of HSV-1 a novel in vitro method was created. In this study HEL-30 keratinocytes were infected with a 0.1 MOI of HSV-1 and plated unto a membrane with 8μm diameter pores. The membrane insert was then placed into culture wells that contained Neuro-2a cells (N2A). The neurites from the neuronal cells made cell-to-cell contact with the infected HEL-30 cells and the virus was transmitted into the neuronal cells. To determine if infection occurred in the N2A cells the cells were lysed and the lysate incubated with Vero cells to titrate virus plaque …

White Blood Cell Counts, Parasite Prevalence, And Plasma Cortisol Levels Of Dogs In A County Animal Shelter: Changes Over Days And Impact Of A Program Of Repeated Human Interaction, Emily S. Dudley

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Animal shelter housing is highly stressful for a dog, compromising welfare and leading to undesirable behaviors and unknown health consequences. We documented the changes in circulating numbers of white blood cells, plasma cortisol, and fecal parasite shedding of dogs housed for 10 days at a county animal shelter. White blood cell changes were most prominent on Day 10 after arrival to the shelter. Changes included increased total leukocytes, mature neutrophils, and lymphocytes, with less consistent increases in monocytes and neutrophil to lymphocyte ratio (N:L). Fecal parasite shedding was elevated and not affected by day. Cortisol levels of shelter dogs declined …

Identification Of The Sites Of Action Of Inhibitors Of Mammalian Phospholipase D2 (Pld2) And The Role Of Interacting Protein Partners, Ramya Ganesan

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Phospholipase D (PLD) is a key enzyme for the remodeling of phospholipids in the cell membrane. PLD has been implicated in many physiological functions such as chemotaxis and phagocytosis as well as pathological functions such as ischemia/reperfusion and cancer metastasis. Several small molecule inhibitors of PLD have been recently developed to overcome these pathological effects. We have concentrated on 5-Fluoro-2-Indolyl des-Chlorohalopemide (FIPI) which is an indole derivative of halopemide that inhibits the two mammalian isoforms PLD1 and PLD2. We also concentrate on N-[2-[1-(3-Fluorophenyl)-4-oxo-1,3,-8-triazaspiro[4.5]dec-8-yl]ethyl]-2-naphthalenecarboxamide (NFOT) and N-[2-(4-oxo-1-phenyl-1,3,8-triazaspiro[4,5]dec-8-yl)ethyl]-2-naphthalenecarboxamide (NOPT) that are specific inhibitors of the PLD2 isoform. In spite of knowing their …

Comparison Between Flow Cytometry And Bead Method In Counting Cd4 And Cd8 T Lymphocytes In Mouse Spleen Cells Suspension, Abdulrahman Abdalla Allabidi

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The flow cytometry is gold standard method to count CD4, CD8 human T lymphocytes, and calculate CD8 to CD4 ratio in HIV patients peripheral blood to stage them and monitor their response to treatment. In this study, a comparison was made by counting CD4and CD8 mouse spleen T lymphocytes by flow cytometry, and a method involving antibody coated latex beads. The bead method yielded results comparable to those obtained by flow cytometry. These results indicate that antibody coated beads are suitable to determine CD4 and CD8 + T lymphocytes counts in situation where flow cytometry is not readily available.

Effect Of Herpes Simplex Virus-1 On Macrophage Cd Marker Expression, Hind Obaid Albeshri

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Macrophages are specialized phagocytic cells derived from the peripheral blood mononuclear cells. Macrophages can assume M1 or M2 cellular states upon activation with specific cytokines. Upon activation, macrophages produce either pro- or anti- inflammatory cytokines and thereby perform their functions of antigen engulfment and debris clearance. Macrophages have several cell surface cluster differentiation (CD) markers including CD80, CD163 and CD200R that differentiate M1 macrophages from M2. Downstream signaling and cytokine production varies based on the interaction between the specific CD marker and antigen.

The present study was aimed at analyzing and comparing the effects of HSV-1 infection on un-polarized and …

Elucidating The Effects Of Tcdd On The Polymorphic Human Hs1,2 Enhancer, Abdullah Freiwan

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2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent environmental toxin known to inhibit immunoglobulin (Ig) gene expression in various animal studies. We have identified the mouse 3'Ig heavy chain regulatory region (3'IgH RR) as a sensitive transcriptional target of TCDD, which may mediate the inhibitory effect of TCDD on Ig expression. Interestingly, the human hs1,2 enhancer is polymorphic and has been associated with a number of autoimmune diseases.

Suggesting a species difference, TCDD inhibited mouse hs1,2 enhancer activation and activated basal human hs1,2 (hs-hs1,2) enhancer activity in the mouse B-cell line. The objective of this study was to elucidate the effects of TCDD …

Adeno-Associated Virus (Aav) Transduction Of Primary Human Cd4+T Lymphocytes, Sahar Hussein Kamel

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Development of latent HIV-infected CD4+ T lymphocytes is the major cause of HIV treatment failure. Adeno-associated virus (AAV) is an attractive vector for anti-HIV gene therapy due to its lack of pathogenicity, low immunogenicity, and persistent transgene expression. However, a major limitation for AAV gene transfer is the cell-specific tropism of each serotype. Only AAV serotypes 2 and 5 have been investigated in hematopoietic cells, both of these serotypes have shown low transduction efficiency. Since each of the currently described AAV serotypes demonstrate distinct tissue tropism, I hypothesized that other serotypes such as AAV1, 4, 8, or 9 may demonstrate …

Expression Of The Alpha, Beta, And Gamma Subunits Of The Interleukin-2 Receptor By Human Vascular Smooth Muscle Cells, Sultan Mohammed Alhayyani

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Interleukin 2 (IL-2) is a member of the cytokine family and contributes to the proliferation, survival, and death of lymphocytes [1]. The interleukin-2 receptor (IL-2) is a tripartite receptor commonly expressed on the surfaces of many lymphoid cells and is composed of three non-covalently associated subunits, alpha (CD25), beta (CD122), and gamma (CD132) [2]. Our laboratory has previously described IL-2 receptor Beta expression by vasculature smooth muscle cells (VSMC) in mice and humans [3]. The current work expands our previous observations by assessing the expression of the alpha and gamma subunits of the IL-2R by VSMCs. Analysis of IL-2R expression …

Single-Tube Real-Time Pcr Assay For Differentiation Of Ixodes Affinis And Ixodes Scapularis, Chelsea L. Wright, Wayne L. Hynes, Breanna T. White, Mindy N. Marshall, Holly D. Gaff, David T. Gauthier

Biological Sciences Faculty Publications

Ixodes affinis Neumann (1899) and Ixodes scapularis Say (1821) are tick vectors of the etiologic agent of Lyme disease, Borrelia burgdorferi sensu stricto. Ixodes affinis and I. scapularis are morphologically very similar, and as they are sympatric in the mid- and south-Atlantic U.S. coastal states, their accurate identification is crucial to studies of disease and vector ecology in this area. This work describes a rapid, single-tube SYBR® Green-based real-time PCR assay for differentiation of I. affinis and I. scapularis at all life stages. The assay employs 2 pairs of species specific primers directed against the internal transcribed spacer 2 (ITS2) …

Prevalence Of Ehrlichia Chaffeensis And Ehrlichia Ewingii In Amblyomma Americanum And Dermacentor Variabilis Collected From Southeastern Virginia, 2010-2011, Chelsea L. Wright, Holly D. Gaff, Wayne L. Hynes

Biological Sciences Faculty Publications

Amblyomma americanum is the most commonly-encountered tick species in southeastern Virginia, representing approximately 95% of the human-biting tick population in this area. Here we investigated the prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in questing Amblyomma americanum and Dermacentor variabilis ticks collected from multiple sites in southeastern Virginia from 2010–2011. Although both Ehrlichia species were detected in Amblyomma americanum, no evidence of either pathogen was found in Dermacentor variabilis. Prevalence of E. chaffeensis varied by location, ranging from 0 – 5.08% among Amblyomma americanum populations. Ehrlichia ewingii prevalence was slightly higher, ranging from 0 – 8.20% among A. …

Ticks And Spotted Fever Group Rickettsiae Of Southeastern Virginia, Robyn M. Nadolny, Chelsea L. Wright, Daniel E. Sonenshine, Wayne L. Hynes, Holly D. Gaff

Biological Sciences Faculty Publications

The incidence of tick-borne rickettsial disease in the southeastern United States has been rising steadily through the past decade, and the range expansions of tick species and tick-borne infectious agents, new and old, has resulted in an unprecedented mix of vectors and pathogens. The results of an ongoing 4-year surveillance project describe the relative abundance of questing tick populations in southeastern Virginia. Since 2009, more than 66,000 questing ticks of 7 species have been collected from vegetation in a variety of habitats, with Amblyomma americanum constituting over 95% of ticks collected. Other species represented included Ixodes scapularis, Dermacentor variabilis …

Characterizing Dsrna Production In Virus-Infected Fish Cells, Amal Brek Aloufi

Theses and Dissertations (Comprehensive)

Viral dsRNA is produced by almost all viruses sometime during their replicative cycle. These viral nucleic acids are potent inducers of both innate and adaptive immune responses, and are therefore considered important immuno-modulators. Previous studies have shown that viruses produce dsRNA when replicating in mammalian cells; however, to date no one has demonstrated viral dsRNA production in virus infected fish cells. Therefore, the goal of this study is to investigate dsRNA production by fish viruses in fish cells, verifying production and performing initial characterization of the dsRNA molecules being produced. Three different rainbow trout cell lines were used in this …

C-Jun N-Terminal Kinases Regulate Adenovirus-Mediated Autophagy And Antigen Presentation, Sarah R. Klein

Dissertations and Theses (Open Access)

Targeted immunotherapy with recombinant, oncolytic adenoviruses is under investigation for the treatment of cancer. Evidence indicates adenoviruses induce autophagy that is required for oncolysis, but the molecular regulation of autophagy in infected cells remains under investigation. Our data suggested the canonical pathway regulating starvation-induced autophagy was not implemented in adenovirus-induced autophagy; however, adenovirus infection triggered phosphorylation of c-Jun N-terminal kinases (JNK) that was essential for autophagy. Adenoviral replication within the host cell elicited JNK pathway activation leading to B cell lymphoma-2 (Bcl-2) phosphorylation. JNK-dependent Bcl-2 phosphorylation stimulated the dissociation of Bcl-2/beclin 1 heterodimers, enabling beclin 1 to initiate autophagy. Moreover, …

Barrier To Autointegration Factor (Baf) Inhibits Vaccinia Virus Intermediate Transcription In The Absence Of The Viral B1 Kinase, Nouhou Ibrahim, April Wicklund, Augusta Jamin, Matthew S. Wiebe

Nebraska Center for Virology: Faculty Publications

Barrier to autointegration factor (BAF/BANF1) is a cellular DNA-binding protein found in the nucleus and cytoplasm. Cytoplasmic BAF binds to foreign DNA and can act as a defense against vaccinia DNA replication. To evade BAF, vaccinia expresses the B1 kinase, which phosphorylates BAF and blocks its ability to bind DNA. Interestingly, B1 is also needed for viral intermediate gene expression via an unknown mechanism. Therefore, we evaluated the impact of B1-BAF signaling on vaccinia transcription. Strikingly, the decrease in vaccinia transcription caused by loss of B1 can be rescued by depletion of BAF. The repressive action of BAF is greatest …

A Supramolecular Strategy To Assemble Multifunctional Viral Nanoparticles, Limin Chen, Xia Zhao, Yuan Lin, Yubin Huang, Qian Wang

Faculty Publications

Using a one-pot approach driven by the supramolecular interaction between β-cyclodextrin and adamantyl moieties, multifunctional viral nanoparticles can be facilely formulated for biomedical applications.

Coccidia Of Gerbils From Mongolia, Ethan Thomas Jensen

School of Biological Sciences: Dissertations, Theses, and Student Research

In this study, gerbils collected in the Mongolia over the summers of 2009-2012 were examined for coccidia. In total, 171 gerbils of three species from 22 localities were examined for coccidia. Coccidian oocysts were identified from 21 gerbils, but those found in 1 of those gerbils were probably pseudoparasites of the host from which they were recovered. From the remaining 20 gerbils, 7 morphotypes of Eimeria and 1 morphotype of Isospora were identified. Four of the 7 morphotypes of Eimeria were attributed to new species which were described in this study. In addition, 10 previously described species of Eimeria were …