No description available.

No description available.

No description available.

Lassa virus (LASV) is a member of the Old World Complex of arenaviruses infecting an estimated 300,000-500,000 people each year primarily in the endemic region of West Africa. Lassa fever (LF), a neglected tropical disease, puts a heavy burden on the communities in which it is endemic. Reportedly, 15%-20% or up to 50% during epidemics succumb to LASV infection. Without U.S. Food and Drug Administration approved diagnostics, therapeutics, or vaccines, knowledge of this disease must advance in order to create affordable and reliable care. Unlike other viral infections, LF does not stimulate a robust humoral immune response. Many in vitro studies have demonstrated the ability of LASV nucleoprotein (NP) and Z protein to interrupt innate immune activation of important cytokines and costimulatory molecules, which can impair the adaptive immune response. To this end, we investigated the antibody and cytokine responses in LF confirmed patients. We then investigated the role NP plays in subverting the innate immune response, suggesting a possible mechanism for humoral immune impairment. Our findings demonstrate a prolonged IgM response and a dysregulated cytokine response in vivo. Additionally, we show LASV NP is capable of modulating the immune system by masking pathogen pattern signals, namely dsRNA, that initiate immune cascades via the Toll Like Receptor 3 pathway. / 1 / Jessica Nicole Hartnett

Human endogenous retroviruses are the remnants of ancient retroviral infections present within our genome. These molecular fossils show similarities with present day exogenous retroviruses but act as typical Mendelian elements that are passed vertically between generations. Despite being repeatedly linked to a number of autoimmune diseases and disorders, no conclusive proof has been identified. Rheumatoid arthritis (RA) is one such disease which has been associated with an increase in HERV expression, compared to controls. In order to elucidate a clear role for HERVs in RA pathogenesis, autoantigens implicated in disease pathogenesis were scanned for sequence homology to retroviral genes. Such epitopes would induce antibodies cross reactive with host proteins, resulting in disease. Short peptides mimicking these regions were synthesised and the prevalence of anti-HERV antibodies was determined in RA patients and disease controls. Additionally, a novel real-time Polymerase Chain Reaction (PCR) assay was developed to accurately quantify levels of HERV-K (HML-2) gag expression, relative to normalised levels of housekeeping gene expression. Both serological and molecular assays showed significant increases in HERV-K (HML-2) activity in RA patients compared to disease controls with CD4+ lymphocytes harbouring the highest activity. The real-time assay was also used to determine whether factors within the synovium could modulate HERVs, resulting in their upregulation. Exogenous viral protein expression and pro-inflammatory cytokines were shown to exert a significant modulatory effect over HERV-K (HML-2) transcription. From this data, it is clear that RA patients have increased levels of HERV-K (HML-2) gag activity compared to controls. Despite this it is likely that factors within the synovium such as exogenous viral expression and pro-inflammatory cytokines also influence HERV-K (HML-2) transcription possibly contributing to a role of bystander activation, i.e. being influenced by external factors, rather than actively contributing to disease processes. The exact role of HERVs in RA pathology remains elusive; however this research proposes several mechanisms by which HERV-K (HML-2) may contribute to disease.

Gene therapy has shown potential in alleviating a wide range of diseases, ranging from viral infections to autosomal diseases. One of the obstacles to gene therapy reaching its full potential is the inadequacy of methods to deliver therapeutic nucleic sequences. Current delivery of gene therapy entails use of viral and non-viral vectors. Viral vectors are however associated with drawbacks such as potential toxicity, high cost and labour-intensive production. Thus non-viral delivery alternatives are being developed in an attempt to overcome difficulties associated with nucleic acid delivery for gene therapy. Virus-like particles are potentially very useful delivery vehicles as their production is simple and cost effective, the particle surface is amenable to modification and the capsid interior can be altered to accommodate a variety of cargoes. One such particle is the recombinant HBV capsid, which comprises a single species of protein and is tolerant of amino acid substitutions on the surface exposed immunodominant loops. This study aimed to enhance the vector-like properties of the HBV virus-like particle by including amino acid substitutions on the particle surface. These substituted residues in turn provided a conjugation site for tropic and immuno evasive moieties. It was found that lysine substitutions resulted in poor conjugation to the capsid surface, whereas substituted cysteine residues resulted in almost all protein-moiety conjugates forming. In order to introduce lysine and cysteine substitutions, a novel method of cloning into the HBV was generated. In doing so, complicated procedures associated with cloning into the immunodominant loop of the HBV capsid have been alleviated. Ligands containing galactose were utilised on the surface of both the HBV capsid and liposomes to confer hepatotropism. The presence of the galactose moieties on the surface of the HBV capsid prevented indiscriminate cellular uptake in cultured cells, however did not improve hepatotropism. Galactose ligands on the surface of liposomes did improve transfection efficiency, however they required a short linker distance between liposome surface and galactose group. The inclusion of galactose in liposome formulations also provided a means to deliver siRNA to the liver of transgenic HBV mice. It is believed that with alterations to the ligand structure, it is possible to provide HBV capsids with hepatotropism in future experimentation. This study demonstrated that the exposed external surface of the HBV capsid is amenable to convenient conjugation, which potentially facilitates immune evasion and conferring of defined tropism.

The effects of vitamin E (VE) and Arginine (ARG) on humoral and cellular immunity in chickens were investigated in two experiments. The humoral immunity was measured by antibody responses to sheep red blood cells (SRBC) and maternal antibody titers to the infectious bursal disease virus (IBDV), while the cellular immunity was studied using the cutaneous basophil hypersensitivity test to phytogemagglutinin (PHA) and by counting subpopulations of T-lymphocytes. We used two levels of ARG: normal (NARG, 1.2% in feed) and high ARG (HARG, additional 0.3% in drinking water or 1% in feed in experiments 1 and 2, respectively); and three levels of VE were given: 40, 80, and 400 IU/kg feed in experiment 1, and 40, 80, and 200 in experiment 2. / HARG improved the antibody response to SRBC compared with NARG ( P<0.01 for experiment 1 and P<0.013 for experiment 2) 4 days after injection in both experiments. In experiment 1, the VE80 birds maintained higher antibody titers to SRBC (P<0.001) than the VE40 and VE400 birds 4, 8 and 16 d after inoculation. In experiment 2, the antibody titers to SRBC were higher in the VE80 birds compared with the VE200 birds at days 5, 8, and 12 after inoculation (P<0.001). Maternal antibody titers (log10) to the IBDV were higher in the HARG than in the NARG diet in 17-day-old birds (P<0.001) and higher in the VE80 than in the VE40 birds (P<0.001), yet similar to those of the VE200 birds. No interactions were found between ARG and VE. / Naive birds fed HARG exhibited a higher response than NARG birds (P<0.05) to PHA-P at d 17 and to PHA-M at d 41, but, after a second exposure, high ARG levels did not have an effect. Also, in naive birds, the effects of VE were not significant at d 17, but showed an influence after a second exposure in 41-d-old birds. / The percentage of T-helper (Th) and T-cytotoxic (Tc) cells in the blood of 29-d-old birds were not different between ARG levels (P=0.07 and P=0.06, respectively), but Th cells were higher in the VE80 and VE200 birds than in the VE40 birds, and Tc was higher in the VE80 than in the VE40 birds (P=0.02). The B-cell:T-cell ratio was higher in the HARG than the NARG birds (P=0.01) and in the VE40 compared with the VE80 and VE200 birds (P<0.001). Neither ARG nor VE had an effect on the ratio of Th:Tc cells, nor on the percentage of immature T-lymphocytes. / A combination of high levels of ARG and high levels of VE (80 IU/kg of BW) has an important immunomodulation effect on the cellular and humoral immune responses in broiler chickens, improving both maternal antibody titers against the IBDV and antibody titers against SRBC. A combination of ARG and VE increases the proportions of Th and Tc cells, the B-cell:T-cell ratio, and growth performance. The evidence suggests that ARG and VE play complementary and regulatory role on immune response and may enhance the resistance of broilers to infectious diseases. / Key words. Arginine, vitamin E, humoral immunity, cell-mediated immunity, lymphocyte, ELISA. / L'effet de la vitamine E (VE) et l'arginine (ARG) sur les systèmes hummoraireet cellulaire de l'immunité a était évalué chez la volaille dans deux recherches. Lesystème hummoraire de l'immunité a était évalué en utilisant les paramètres tels que laproduction d'anticorps après une injection des globules rouge provenant des moutons(SRBC) et le niveau d'anticorps maternelle après une infection avec les virus causantla maladie 'infectious bursal disease' (lBDV), tandis que les effets sur le systemcellulaire de l'immunité avaient aussi été évalués en utilisant les paramètres comme'cutaneous basophil hypersensitivity test to phytogemagglutinin (PHA)' et endéterminant la concentration des lymphocytes T. Deux concentrations de ARG avaientété utilisées: normale (NARG, 1.2 % de la diète) et une concentration élevée (HARG,additionel 0.3 % dans l'eau ou 1 % dans les diètes); et 3 concentrations de VE: 40, 80et 400 lU/kg dans les diètes dans la première recherche et 40, 80, et 200 lU/kg dans ladeuxième recherche.

Thesis (M.Sc.) Uppsala : Sveriges lantbruksuniv. / Härtill 1 uppsatser.

No description available.

Understanding the host response immediately following mucosal HIV-1 infection will be pivotal in determining whether the immune response induced by a vaccine will successfully sense and control viral replication. In order for effective vaccine strategies and modalities to be developed, these earliest immunological events must be fully assessed in a non-biased manner. Nonhuman primates (NHP), specifically Rhesus macaques (RM), serve as a model to investigate the immunological landscape immediately post-challenge and to define the spatiotemporal path of simian immunodeficiency virus (SIV). SIV infection of RM serves as a model of human HIV infection as it recapitulates many of the virological, immunological, and pathological features of HIV infection in the human host. In this thesis I will test the hypothesis whether transcriptional analysis will allow a sensitive measure of the early innate immune responses that accompany detection of the SIV virus in the periphery. I have determined that an early inflammatory profile arises early in tissues proximal to the challenge site that precedes widespread immune activation and the systemic antiviral interferon response. This study defines in detail the spatiotemporal relationship between virus and host immune response and may be a valuable resource in guiding future vaccine design strategies. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2016. / FAU Electronic Theses and Dissertations Collection