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Expression Of Sars Cov2 Receptors Influenced Upon Cytokine Polarizations (Il-4 And Ifnγ) In Hemangioendothelioma Cells, Chandra Lekha Koopari Jan 2022

Expression Of Sars Cov2 Receptors Influenced Upon Cytokine Polarizations (Il-4 And Ifnγ) In Hemangioendothelioma Cells, Chandra Lekha Koopari

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Endothelial cells are distinct multifunctional cells with essential metabolic and synthetic roles along with their ability to function as selective permeability barrier. Endothelial cells (ECs), the major component of blood vessels, essentially interact directly yet differently with inflammatory cytokines. ECs are well recognized to be polarized cells, but little is known about the potential function of inflammatory mediators. Covid 19 may have long-term health effects on par with chronic illnesses. Vascular inflammatory disease and coagulopathy linked to COVID-19 are exacerbated by endothelial cell (EC) dysfunction. SARS-CoV-2 enters the host cell through the Angiotensin Converting Enzyme 2 (ACE2) receptor and primes …


Ifn-Γ Increases The Expression Of Sars-Cov-2 Receptors On Vero E6 Cells, Bindu Madhavi Madabattula Jan 2022

Ifn-Γ Increases The Expression Of Sars-Cov-2 Receptors On Vero E6 Cells, Bindu Madhavi Madabattula

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Respiratory epithelial cells are the initial target for SARS-CoV-2 infection. IFN-γ is known to increase the expression of ACE-2, an initial receptor for SARS-CoV-2, on epithelial cells. This study focuses on examining the effect of IFN-γ for ACE-2, TMPRSS2, and neuropilin-1 on Vero E6 cells using two immunofluorescence methods, namely, direct (membrane) fluorescence method and Cytation5 method. Direct fluorescence was determined using an Accu-Scope and ImageJ analysis. Using this method, significance (p<0.023) was observed only for ACE-2 when Vero E6 cells were treated with IFN-γ. Cytation5 fluorescence was determined using a Bio-tek Cytation5 plate reader. The results showed that IFN-γ significantly increased (p<0.001) the expression of ACE-2, neuropilin-1, and TMPRSS2. These results indicate Cytation5 is a more sensitive method for determining the expression of receptors on Vero E6 cells. The elevated levels of SARS-CoV-2 receptors expression resulting from IFN-γ treatment makes the epithelial cells more susceptible targets of SARS-CoV-2 infection. IFN-γ is most likely provided by innate immune cells in the initial COVID-19 infection, consequently contributing to the severity of disease.


Il-10 And Tgf-Beta Increase Connexin-43 Expression And Membrane Potential Of Hl-1 Cardiomyocytes Coupled With Raw 264.7 Macrophages, Cora B. Cox Jan 2021

Il-10 And Tgf-Beta Increase Connexin-43 Expression And Membrane Potential Of Hl-1 Cardiomyocytes Coupled With Raw 264.7 Macrophages, Cora B. Cox

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Cardiomyocytes and macrophages have been found to interact via connexin-43 hemichannels. The role of connexin-43, however, is not fully understood. This study shows that these interactions aid in increasing the membrane potential of cardiomyocytes allowing contraction of the cells. HL-1 cardiomyocytes and RAW 264.7 macrophages in coculture increased expression of connexin-43 compared to cardiomyocytes alone. Co-cultures also increased the fluorescence of Di-8-ANEPPS potentiometric dye indicating an increase in cardiomyocyte membrane potential. Treatment with IL-10 and TGF-beta further increased connexin-43 expression and membrane potential. Treatment with SOCS3 inhibited the effects of TGF-beta and IL-10 while having no effect on its own. …


Functional Characterization Of Infectious Hematopoietic Necrosis Virus Matrix Protein In Host Cellular Responses, Jeffery Ringiesn Jan 2021

Functional Characterization Of Infectious Hematopoietic Necrosis Virus Matrix Protein In Host Cellular Responses, Jeffery Ringiesn

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Infectious hematopoietic necrosis virus (IHNV) is a deadly fish pathogen that poses a global threat to aquatic ecosystems and the aquaculture industry. For decades, research has focused on developing vaccine therapeutics utilizing a variety of techniques and strategies. While these studies have met with some success in identifying potential vaccine targets that provided protective immunity, a commercially viable IHNV vaccine is currently unavailable. Here we explore the relationship between the structure and function of the IHNV matrix (M) protein through the introduction of mutations that reduce anti-host effects, with the goal of developing a novel recombinant IHNV with reduced pathogenicity …


Divalent Metal Cation Entry And Cytotoxicity In Jurkat T Cells: Role Of Trpm7 Channels, Alayna N. Mellott Jan 2020

Divalent Metal Cation Entry And Cytotoxicity In Jurkat T Cells: Role Of Trpm7 Channels, Alayna N. Mellott

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Humans are exposed daily to a variety of metals that can be harmful to our immune system. Although certain divalent metal cations are essential for numerous cellular functions and are critical trace elements in humans, the uptake mechanisms of these ions remain mostly unknown. Transient receptor potential melastatin 7 (TRPM7), which is expressed in a variety of human cell types, including lymphocytes and macrophages, conducts many divalent metal cations. TRPM7 channels are largely inactive under normal physiological conditions due to cytoplasmic magnesium acting as a channel inhibitor. Magnesium is a cofactor for many biochemical reactions. Low serum levels of magnesium, …


The Persisting Threats Of Cholera: A Cyclical Public Health Problem In Ghana, Rita Laryea Amediavor Jan 2020

The Persisting Threats Of Cholera: A Cyclical Public Health Problem In Ghana, Rita Laryea Amediavor

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The prevalence of communicable diseases continues to be one of the continent's leading causes of deaths. Cholera is a waterborne disease triggered by toxigenic strains of the Gram-negative bacteria Vibrio Cholerae O1 strain and less common O139 strain. with symptoms such as severe acute watery diarrhea and vomiting leading to dehydration, progressing to hypovolemic shock and death if not treated timely due to its short incubation period (Pasetto et al., 2018). West African countries are largely portrayed as endemic to cholera, though the dynamics of outbreaks in these developing countries remain largely uncertain. The purpose of the study is to …


Potential Role Of Ahr In Antibody Production, Mili Bhakta Jan 2020

Potential Role Of Ahr In Antibody Production, Mili Bhakta

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Aryl hydrocarbon receptor (AhR) mediates the immunosuppressive effects of 2,3,7,8 -tetrachlorodibenzodioxin (TCDD) in murine B cells. The effects of AhR activation on the regulation of expression of human immunoglobulin isotypes (μ, γ1-4, α1-2 and ε) and Ig secretion is unclear. Our previous results using CL-01 cell-line originating from a Burkitt’s lymphoma patient, demonstrated an inhibitory effect of TCDD on IgG expression but a surprising and marked loss of IgG secretion when the AhR was knocked out by siRNA or CRISPR/Cas9 gene editing. To determine if the AhR is a critical mediator of IgG expression, current study is focused on characterizing …


The Response Of M0, M1, And M2 Raw246.7 Macrophage Cell Line To Hsv-1 Infection In Vitro, Amani Mohammed Alhazmi Jan 2019

The Response Of M0, M1, And M2 Raw246.7 Macrophage Cell Line To Hsv-1 Infection In Vitro, Amani Mohammed Alhazmi

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Herpes Simplex Virus Type 1 (HSV-1) infection occurs through the epithelial cells of the skin or mucous membranes. The beginning of the primary infection is rapid and is characterized by pain in the mouth, salivation, and submandibular lymphadenitis. The infected mucosa produces numerous, small and red lesions known as cold sores, however, many cases are asymptomatic. After the primary infection HSV-1 moves through the nerve to stay in trigeminal ganglia and to cause a recurrent infection from time to time. In the early hours of the HSV-1 infection, the cytokines produced by infected cells are critical in the stimulation of …


Six-Nine Months Long Term Culture Of Mouse Bone Marrow Cells Differentiated To Macrophages And Eosinophils, Olena B. Svitlova Jan 2019

Six-Nine Months Long Term Culture Of Mouse Bone Marrow Cells Differentiated To Macrophages And Eosinophils, Olena B. Svitlova

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Mouse models of eosinophil-associated diseases have been used to study the mechanisms of disease pathogenesis. In this study, mouse-derived bone marrow cells were used in long-term (6 and 9 months) cell cultures of differentiated eosinophils and macrophages. IL-5 was used to differentiate the stem cells to eosinophils and GM-CSF was used to propagate macrophages from the bone marrow stem cells. The maximum time period for observing the eosinophil cultures was 252 days which is censurably longer than the 18 days culture period observed by others. The results were assessed by describing the microscopic cell morphology by Wright staining, modified Giemsa …


Enhanced Expression Of Receptor Tyrosine Kinase Mer (Mertk) On Socs3-Treated Polarized Raw 264.7 Anti-Inflammatory M2c Macrophages, Sankhadip Bhadra Jan 2019

Enhanced Expression Of Receptor Tyrosine Kinase Mer (Mertk) On Socs3-Treated Polarized Raw 264.7 Anti-Inflammatory M2c Macrophages, Sankhadip Bhadra

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Macrophages are phagocytic cells located in tissues, organs and even circulated within our body as white blood cells. They are critical in detecting tissue damage and infection. Resident tissue macrophages initiate the signals for inflammation recruiting neutrophils and blood monocytes which mature into macrophages at sites of infection and in the resolution of inflammation. Based on the local cytokine milieu in tissue sites, macrophages may be polarized into pro-inflammatory M1 or anti-inflammatory M2 phenotypes. Receptor tyrosine kinase Mer (MERTK) helps in clearing dead neutrophils and other apoptotic cells from damaged tissue sites preventing chronic inflammation and autoimmune disorders. MERTK aids …


The Effects Of Socs1 And Socs3 Peptide Mimetics On Macrophage Phagocytosis Of Malignant Cells, Tahirah M. Madkhali Jan 2019

The Effects Of Socs1 And Socs3 Peptide Mimetics On Macrophage Phagocytosis Of Malignant Cells, Tahirah M. Madkhali

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Macrophages are essential phagocytic cells involved in both innate and adaptive immune systems and play vital roles in the host defense and inflammation. Macrophages have a remarkably high capacity to clear unnecessary cellular materials in interstitial environment through a process called “phagocytosis”, which is affected by many factors including suppressors of cytokine signaling (SOCS). SOCSs are a group of intracellular proteins that downregulate the cytokine signals involved in various JAK/STAT pathways through a negative feedback loop. This study focuses on investigating the effects of SOCS1 and SOCS3 on the phagocytic ability of RAW 264.7 macrophages polarized into M2a with IL-4/IL-13 …


Virus Production And Cell Viability Of Hsv-1-Infected Murine Keratinocytes (Hel-30) Co-Cultured With Murine Macrophages (Raw 264.7), Barry Graffagna Jan 2018

Virus Production And Cell Viability Of Hsv-1-Infected Murine Keratinocytes (Hel-30) Co-Cultured With Murine Macrophages (Raw 264.7), Barry Graffagna

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Keratinocytes are the most abundant type of cell in the outer layer of skin, the epidermis, and provide barrier against pathogens from invading. However, Herpes Simplex Virus Type 1 (HSV-1) targets these keratinocytes for infection, and later infects neurons to establish lifelong latency. The keratinocytes stimulate the innate immune system to engage and to destroy the virus. Among the cells of the innate immune system to respond to the viral invasion is the macrophage. In this study, RAW 264.7 macrophage and HEL-30 keratinocyte monolayers were challenged in vitro with HSV-1 at a multiplicity of infection (MOI) of 0.1 to investigate …


The Impact Of Socs1 And Socs3 Peptide Mimetics On Rho And Cdc42 Proteins Expression, F-Actin Cytoskeleton Rearrangements, And Cytokines Production Of Uninfected And Hsv-1 Infected M1 And M2 Raw 264.7 Murine Macrophages, Maha A. Elwardany Jan 2017

The Impact Of Socs1 And Socs3 Peptide Mimetics On Rho And Cdc42 Proteins Expression, F-Actin Cytoskeleton Rearrangements, And Cytokines Production Of Uninfected And Hsv-1 Infected M1 And M2 Raw 264.7 Murine Macrophages, Maha A. Elwardany

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The immune system plays an essential role in the pathogenesis of HSV-1 during the lytic phase of the disease, orchestrating the inflammatory response, retaining the virus in its latent phase and preventing the recurrence of HSV-1 infection. Macrophages display a vital role in the innate and adaptive immune responses during multiple phases of HSV-1 infection. Polarized macrophages are categorized into two distinct classes with diverse functions. The classically activated M1 can engulf and destroy the microbial agents, produce proinflammatory cytokines, and participate in the pathogenesis of many inflammatory diseases. The alternatively activated M2 induces anti-inflammatory mediators and stimulates tissue remodeling …


Effect Of Exposure Of Raw264.7 Macrophages To Salmonella Typhimurium Components On Cell Viability, Cytoskeleton Re-Arrangement And Cytokine Secretion, Khalid Abdullah Alyahya Jan 2017

Effect Of Exposure Of Raw264.7 Macrophages To Salmonella Typhimurium Components On Cell Viability, Cytoskeleton Re-Arrangement And Cytokine Secretion, Khalid Abdullah Alyahya

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Innate immune system plays an important role in individual's protection against pathogens and in activation of adaptive immune system. Utilizing RAW 264.7 murine macrophages as an innate immune response representative in this study, we analyzed the effect of invasive pathogen's components (e.g. flagellin) on the arrangement of macrophage's cytoskeleton, on viability of immune cells and on secretion of pro-inflammatory and anti-inflammatory cytokines and on fluorescence intensity of cytoskeleton after rearrangement. Additionally, we studied the similarity and differences between bacterial (Salmonella typhimurium) and synthetic TLR4 agonist (synthetic lipid-A) on viability, fluorescence intensity, cytokine secretion, and cytoskeleton rearrangements. Similarly, we studied the …


The Effect Of Gram-Positive Staphylococcus Aureus Cell Wall Components Lipoteichoic Acid And Peptidoglycan On Cytokine Production, Cytoskeletal Arrangement, And Cell Viability On Raw 264.7 Murine Macrophages, Gabrielle String Jan 2017

The Effect Of Gram-Positive Staphylococcus Aureus Cell Wall Components Lipoteichoic Acid And Peptidoglycan On Cytokine Production, Cytoskeletal Arrangement, And Cell Viability On Raw 264.7 Murine Macrophages, Gabrielle String

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In this study, gram positive Staphylococcus aureus cell wall components such as lipoteichoic acid (LTA) and peptidoglycan (PGN) were used to study the potential inflammatory and anti-inflammatory cytokine response, cytoskeletal arrangement and cell viability on RAW264.7 murine macrophages over 24 hours. The effect of S.aureus LTA and PGN (5 µg/mL) on RAW 264.7 macrophages was evaluated every six hours for twenty-four hours. Inflammatory cytokine (TNF-a) production peaked at 6 hours before decreasing over time. Anti-inflammatory cytokine (IL-10) production peaked between 12 and 18 hours. During the first twenty-four hours, cytotoxicity of treated macrophages, as defined as the release of lactate …


The Effect Of Gram-Positive Staphylococcus Aureus Cell Wall Components Lipoteichoic Acid And Peptidoglycan On Cytokine Production, Cytoskeletal Arrangement, And Cell Viability On Raw 264.7 Murine Macrophages, Gabrielle String Jan 2017

The Effect Of Gram-Positive Staphylococcus Aureus Cell Wall Components Lipoteichoic Acid And Peptidoglycan On Cytokine Production, Cytoskeletal Arrangement, And Cell Viability On Raw 264.7 Murine Macrophages, Gabrielle String

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In this study, gram positive Staphylococcus aureus cell wall components such as lipoteichoic acid (LTA) and peptidoglycan (PGN) were used to study the potential inflammatory and anti-inflammatory cytokine response, cytoskeletal arrangement and cell viability on RAW264.7 murine macrophages over 24 hours. The effect of S.aureus LTA and PGN (5 [mu]g/mL) on RAW 264.7 macrophages was evaluated every six hours for twenty-four hours. Inflammatory cytokine (TNF-a) production peaked at 6 hours before decreasing over time. Anti-inflammatory cytokine (IL-10) production peaked between 12 and 18 hours. During the first twenty-four hours, cytotoxicity of treated macrophages, as defined as the release of lactate …


Cell Viability, Cytoskeleton Organization And Cytokines Secretion Of Raw 264.7 Macrophages Exposed To Gram-Negative Bacterial Components, Ali Awadh Alshehri Jan 2016

Cell Viability, Cytoskeleton Organization And Cytokines Secretion Of Raw 264.7 Macrophages Exposed To Gram-Negative Bacterial Components, Ali Awadh Alshehri

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Macrophages play an important role in innate immunity by controlling cellular responses. In this study, the effects of gram-negative bacterial components (Flagellin, lipoprotein, lipopolysaccharides (LPS), outer membrane proteins-A (OMP-A) and peptidoglycan) were determined on cell viability, morphology, cytoskeletal filament and cytokines secretion of murine RAW 264.7 macrophages at 24 hours. The effect of LPS, flagellin and peptidoglycan from gram negative bacteria on viability murine RAW 264.7 macrophages were evaluated using different concentrations (1, 5 and 10 µg/ml). Cells stimulated with LPS displayed ~ 2-fold decrease (P=0.001) in cell viability compared to control cells at 24 hours whereas cells stimulated with …


The Expression Of Aryl Hydrocarbon Receptor In Raw 264.7 Macrophages In The Presence Of Socs1 Peptide And Socs3 Peptide Mimetic And Cells Infected With Hsv-1, Maher Salem Alwethaynani Jan 2016

The Expression Of Aryl Hydrocarbon Receptor In Raw 264.7 Macrophages In The Presence Of Socs1 Peptide And Socs3 Peptide Mimetic And Cells Infected With Hsv-1, Maher Salem Alwethaynani

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Macrophages play a crucial role for our immune system and protect our body from infection. Suppressor of cytokine signaling (SOCS) proteins negatively regulate cytokine receptor and TLRs. The aryl hydrocarbon receptor (AhR) also performs an important role in immunity. This study investigated the changes in expression of AhR in RAW 264.7 macrophage cells after the addition of SOCS1 and SOCS3 peptide mimetics and also examined AhR expression in RAW 264.7 macrophage cells before and after the addition of HSV-1 RAW 264.7 murine macrophage cell lines which are from male BALB/c mice were used in this study. The addition of the …


Identification Of Protein-Protein Interactions Of Amyotrophic Lateral Sclerosis Associated Protein Tdp-43, Hanoor Sharma Jan 2016

Identification Of Protein-Protein Interactions Of Amyotrophic Lateral Sclerosis Associated Protein Tdp-43, Hanoor Sharma

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Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease caused by progressive degeneration of upper and lower motor neurons in the brain and spinal cord. Multiple mutations are found in some of the proteins associated with ALS, including superoxide dismutase (SOD1), fused in sarcoma (FUS) and trans-activation response DNA-binding protein (TDP-43). TDP-43 is a DNA and RNA binding protein, well conserved, and ubiquitously expressed in all tissues. TDP-43 resides in the nucleus and sometimes shuttles between nucleus and cytoplasm. Mutations in TDP-43 leads to mislocalization of TDP-43 to the cytosol where it was ubiqutinated and hyperphosphsorylated, ultimately leading to neuronal cell …


Changes In Cytoskeleton Proteins In Hsv-1 Infection Of J774a.1 Macrophage Phenotype, Riham Abbas Subahi Jan 2016

Changes In Cytoskeleton Proteins In Hsv-1 Infection Of J774a.1 Macrophage Phenotype, Riham Abbas Subahi

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The cell cytoskeleton, a unique intracellular matrix found in all eukaryotes, is composed of three main protein structures, microtubules, microlaments and intermediate laments. The cytoskeleton maintains cell shape and internal structures providing mechanical support that facilitates intracellular transport (Parker et al., 2014). Many viruses such as the herpesviruses use the cytoskeleton system of the cell for infectivity (Henry Sum, M. S. 2015). HSV-1 utilizes the cell cytoskeleton in many steps of its life cycle from entry through assembly to egress (Lyman et al., 2008). During infection, HSV-1 viral proteins cause drastic changes and rearrangement of the cellular actin and cell …


Species Differences In The Effects Of Tcdd On A Transcriptional Regulatory Region Within The Ig Heavy Chain Gene, Zahra J. Alfaheeda Jan 2016

Species Differences In The Effects Of Tcdd On A Transcriptional Regulatory Region Within The Ig Heavy Chain Gene, Zahra J. Alfaheeda

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The immunoglobulin heavy chain gene (Igh) is regulated by numerous regulatory elements including the 3'Igh regulatory region (3'IghRR). Several transcription factors are involved in modulating the 3'IghRR including NF-B, AP-1, and the aryl hydrocarbon receptor (AhR). The AhR is a ligand-activated transcription factor that mediates the transcription of genes involved in the metabolism of environmental toxicants such as TCDD. TCDD binds AhR and regulates immunoglobulin (Ig) expression in B cells of animal models. This modulation appears to be directly mediated by binding of the AhR to dioxin response elements (DRE) within the 3'IghRR. Structural differences have been found between the …


The Impact Of Hsv-1 Infection On Cell Viability, Morphology, And Cd Markers Expression By Unpolarized And Cytokine-Polarized J774a.1 Mouse Macrophages, Sarah Saad Alsharif Jan 2015

The Impact Of Hsv-1 Infection On Cell Viability, Morphology, And Cd Markers Expression By Unpolarized And Cytokine-Polarized J774a.1 Mouse Macrophages, Sarah Saad Alsharif

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Macrophages play an important role in the immune system, particularly in neutralizing pathogens via phagocytosis and the production of multiple cytokines and chemokines that control infection after exposure to specific stimuli. Macrophages exhibit two different phenotypes, M1 and M2. This study evaluated the role of Suppressors of cytokine signaling (SOCS)1 and SOCS3 on Herpes Simplex Virus (HSV-1) infection of polarized macrophages, cell viability, cell morphology, and the expression of cell surface CD markers. I hypothesized that J774A.1 murine macrophages cells in the naive state (M0), and M1 and M2 phenotypes would display differences in CD markers CD80,CD163, and CD200R, cell …


The Use Of Antibody-Coated Latex Beads To Determine Single Positive And Double Positive Mouse Spleen Cells Expressing Cd5 And/Or Cd19 Glycoproteins, Abdulrazzag Abdulaziz Othman Jan 2015

The Use Of Antibody-Coated Latex Beads To Determine Single Positive And Double Positive Mouse Spleen Cells Expressing Cd5 And/Or Cd19 Glycoproteins, Abdulrazzag Abdulaziz Othman

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Flow cytometry is the standard method used to diagnose, stage, and monitor patients' response to the treatment given by counting the numbers of CD5, CD19 and CD5+ CD19+ B lymphocytes. In this study, a comparison was done between numbers of single CD5+, single CD19+ and dual CD5+ CD19+ mouse spleen B lymphocytes using flow cytometry and antibody-latex beads. The bead method involved antibody-coated latex bead and yielded results similar to those of flow cytometry. For cells exhibiting both markers (CD5+ CD19+), the bead method used antibody-coated beads of two different colors yielded similar results to those of flow cytometry results. …


Socs1/Socs3 Expression And Virus Replication Of Denv2 And Hsv-1 In Cytokine-Polarized Subsets Of Raw 264.7 Macrophages, Nagarjuna Reddy Cheemarla Jan 2014

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 …


Early Increase Of Cd11c In Human Monocyte-Derived Dendritic Cells In The Presence Of A/California/07/2009 (H1n1pdm), Amber M. Braddock Jan 2014

Early Increase Of Cd11c In Human Monocyte-Derived Dendritic Cells In The Presence Of A/California/07/2009 (H1n1pdm), Amber M. Braddock

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Influenza A continues to cause significant morbidity and mortality. Vaccine production time, viral antigenic variability and low vaccine efficacy contribute to the annual pandemics and seasonal epidemics. A better understanding of human immune responses to influenza A could result in more effective vaccines. Dendritic cells (DCs) play an important role in the initiation of immune responses. Populations of B lymphocytes secrete protective antibodies but the role of different populations of B cells in the protective response is unknown. In this thesis we developed assays to determine subclasses of influenza specific antibodies. Mononuclear cells were assessed using flow cytometry to identify …


Expression Of The Alpha, Beta, And Gamma Subunits Of The Interleukin-2 Receptor By Human Vascular Smooth Muscle Cells, Sultan Mohammed Alhayyani Jan 2014

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 …


Comparison Between Flow Cytometry And Bead Method In Counting Cd4 And Cd8 T Lymphocytes In Mouse Spleen Cells Suspension, Abdulrahman Abdalla Allabidi Jan 2014

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.


Novel Approaches For The Eradication Of Hiv Latently Infected Cells, Sally Al Ali Jan 2013

Novel Approaches For The Eradication Of Hiv Latently Infected Cells, Sally Al Ali

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The development of a suitable experimental cell model to study HIV latency in primary cells could have a massive effect on the current approaches to eradicate virus in latently infected cells. The main proposal of this paper is to develop an in vitro HIV cell model that represents HIV latency in vivo, then to create a more effective viral vector in order to target HIV reservoirs. For this goal, a directed evolution method is suggested to be used in order to mutate the AAV cap gene to generate a recombinant AAV vector that is capable of infecting primary resting CD4+ …


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 Jan 2012

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 …


Use Of Phage Display Libraries To Select For B-Cell Receptor-Specific Peptides Of Chronic Lymphocytic Leukemia Cells, Richard M. Chou Jan 2012

Use Of Phage Display Libraries To Select For B-Cell Receptor-Specific Peptides Of Chronic Lymphocytic Leukemia Cells, Richard M. Chou

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Peptides with high affinity to the B-cell receptor (BCR) fused to a toxin could be an effective therapy for Chronic Lymphocytic Leukemia (CLL) patients. We screened captured BCR of a CLL patient with peptides from 7 and 12-mer phage display libraries, using two strategies. Lymphocytes from two patients diagnosed with CLL expressing two different unmutated VH genes (VH 1-3 and VH4-34, respectively) were used. Membrane BCRs were obtained from patient CLL cells by lysis, identified by western blot, semi-quantified and screened with phage libraries. The first strategy involved using patient VH4-34 BCRs which were captured using goat anti-human IgM to …