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B. abortus is an intracellular facultative bacterial pathogen which causes abortion in cattle and undulant fever in humans. Cattle vaccines such as B. abortus strains 19 and RB51 are live vaccine strains which protect approximately 75% of the vaccinated animals. No effective vaccines are available for the prevention of brucellosis in humans. We are developing vaccinia virus recombinants expressing various B. abortus proteins to prevent brucellosis in susceptible mammalian species. In this work the B. abortus groEL gene encoding the antigenic heat shock protein GroEL was subcloned into vaccinia virus via homologous recombination. Expression of the GroEL protein in vaccinia infected cells in-vivo was confirmed by immunoblotting. Groups of 5 female BALB/C mice were injected with the vaccinia recombinant or appropriate positive and negative control vaccines. Mice were bled and their humoral immune responses assessed. In addition, mice were challenged with virulent B. abortus strain 2308 and protection measured by the rate of splenic clearance of live Brucella. In spite of demonstrating specific GroEL antibodies in recombinant vaccinia injected mice, no significant level of protection was demonstrable. Preliminary lymphocyte transformation assays were carried out to establish if a cell mediated immune response to GroEL was induced in the vaccinated animals. / Master of Science

The attenuated total reflection (ATR) due to excitation of surface plasmon is usually applied to study the dielectric coefficient. In this study the surface plasmon has been excited on the interface between the liquid crystal and silver in Krestchmann configuration. We can obtain the effective reflection index and the tilt-angle of liquid crystal anchored on the interface with the excitation condition of surface plasmon. We also discuss the variation of the tilt-angle of liquid crystal with the various applied bias.

Organosilanes are widely used to bond organic materials such as polymers to inorganic materials in polymer composites. However, the mechanism of adhesion is poorly understood. One postulated mechanism is the interdiffusion of the silane and polymer, along with condensation of the silane to form an interpenetrating polymer network (IPN). The techniques of attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and Raman confocal microscopy have been used to study the diffusion, hydrolysis and condensation of three organosilanes in silicon / PVC / silane laminates. These processes are thought to contribute to the formation of the IPN, and hence to be vital for adhesion. The organosilanes studied were [3-(amino)propyl]trimethoxysilane, also known as A1110, [3-(phenylamino)propyl]trimethoxysilane, known as Y9669, and [3-(mercapto)propyl]triethoxysilane, known as A1891.ATR-FTIR was shown to be an excellent technique for studying the kinetics of silane diffusion through PVC films. It was shown that at room temperature, no diffusion through unplasticised PVC films occurred. At 70 °C, however, diffusion occurred readily for Y9669 and A1891. In plasticised PVC films, diffusion was observed for all three silanes at room temperature. It was shown that the diffusion occurred more quickly with higher plasticiser concentrations, and hence lower glass transition temperatures. The kinetics of diffusion was found to fit a dual mode sorption model. Hydrolysis of the silanes was also followed by infrared spectroscopy, and the kinetics of hydrolysis and condensation were shown to be highly dependent upon silane type, the concentration of water, and the presence of an acid catalyst. The hydrolysis of the silanes was found to slow their diffusion through both plasticised and unplasticised PVC films. It was shown that the presence of water in the films caused the hydrolysis of the silanes in situ. Raman depth profiles were measured of the films before, during and after diffusion. The spatial resolution was shown to be adversely affected by refraction at the air / PVC interface. It was shown that it is possible to deconvolve the confocal response of the microscope from the depth profiles, resulting in greater spatial resolution. Hydrolysis of the silanes was followed in solution by Raman spectroscopy, and it was found that each of the three silanes showed different rates of hydrolysis and condensation. It was shown that it was also possible to follow the kinetics of diffusion by Raman microscopy, and the results agreed well with those shown by ATR-FTIR spectroscopy.

In the present dissertation, we characterize the range of the attenuated Radon transform of zero, one, and two tensor fields, supported in strictly convex set. The approach is based on a Hilbert transform associated with A-analytic functions of A. Bukhgeim. We first present new necessary and sufficient conditions for a function to be in the range of the attenuated Radon transform of a sufficiently smooth function supported in the convex set. The approach is based on an explicit Hilbert transform associated with traces of the boundary of A-analytic functions in the sense of A. Bukhgeim. We then uses the range characterization of the Radon transform of functions to characterize the range of the attenuated Radon transform of vector fields as they appear in the medical diagnostic techniques of Doppler tomography. As an application we determine necessary and sufficient conditions for the Doppler and X-ray data to be mistaken for each other. We also characterize the range of real symmetric second order tensor field using the range characterization of the Radon transform of zero tensor field.

The properties of human immunodeficiency virus type 1 (HIV-1) and its simian counterpart SIV that enable persistent replication in the face of robust cellular, antibody, and innate immune responses have complicated efforts to develop a safe and effective vaccine. Vaccine protection against HIV-1 infection may require a combination of immune mechanisms. However, the types of immune responses that can be induced by vaccination to prevent HIV-1 infection remain unclear. The features of the viral envelope glycoprotein (Env) that confer inherent resistance to neutralization by antibodies also interfere with the development of antibody responses. We therefore vaccinated rhesus macaques with single-cycle SIV (scSIV) strains expressing Env proteins mutated to remove features that interfere with the induction of antibody responses. Antibodies capable of neutralizing Env-modified but not wild-type SIV were selectively enhanced. Identifying the immune responses underlying complete protection by live-attenuated SIV against pathogenic SIV challenge may provide guidance for HIV-1 vaccine design. To test the hypothesis that antibodies not measurable by assays for virus neutralization correlate with protection by live-attenuated SIV, we developed a novel assay for antibody-dependent cell-mediated cytotoxicity (ADCC). ADCC activity increased progressively over time after inoculation, and was measurable against viruses expressing heterologous Env proteins from independent SIV isolates when neutralization was undetectable. Two separate pathogenic \(SIV_{mac}251\) challenge experiments took advantage of either the strain specificity or the time-dependent development of immunity to overcome complete protection by live-attenuated SIV. In both experiments, macaques inoculated with live-attenuated SIV that remained uninfected by \(SIV_{mac}251\) had significantly higher ADCC activity than those that became infected. We also measured ADCC for the primary immune correlates analysis of a recent HIV-1 vaccine clinical trial in Thailand (RV144) that reported modest vaccine protection (31%). There was a nonsignificant trend towards lower risk of infection among vaccinees with high versus low relative ADCC activity. However, Env-specific IgA correlated with risk, prompting an analysis stratified by IgA levels. Among vaccinees with low Env-specific IgA, there was lower risk of infection among those with higher ADCC activity. These observations suggest that antibodies that direct ADCC may contribute to vaccine protection against SIV and HIV-1 infection.

Edwardsiella ictaluri causes enteric septicemia of catfish (ESC), one of the most important bacterial diseases of farmed channel catfish in the USA. Use of live attenuated vaccines (LAVs) is an effective strategy for combating mortalities in catfish farms. Our research group has developed three live attenuated E. ictaluri strains [EiΔevpB, EiΔgcvPΔsdhCΔfrdA (ESC-NDKL1), and EiΔhemRΔfrdAΔsdhC (triple-hemR)] that provide various levels of protection against ESC. However, the protective mechanisms of these vaccine candidates are mostly unknown. The overall objective of my study was to investigate protective mechanisms of these LAVs. To accomplish this, catfish fry were immersion challenged with EiΔevpB, ESC-NDKL1, and triple-hemR. Additional catfish fry were immersion challenged with Aquavac-ESC and E. ictaluri wild-type (EiWT) as controls. The internalization of antigens through the mucosal surfaces as well as the pathology and molecular immune responses were studied. The investigations showed that EiΔevpB and ESC-NDKL1 were highly safe and efficacious compared to Aquavac-ESC and EiWT. Pathologically, vaccination with EiΔevpB and ESC-NDKL1 decreased the pathological lesions, EiWT replication in catfish, and increased the ability of the immune system to resist and kill EiWT. On the other hand, triple-hemR was not safe causing severe tissue damage similar to EiWT. The gene expression data showed significantly high expression of innate and adaptive immune genes following vaccination and challenge with EiWT. Additionally, EiΔevpB and ESC-NDKL1 induced immune proliferation in pronephros after exposure to EiWT. Taken together, EiΔevpB and ESC-NDKL1 vaccine strains performed better to induce immune responses and reduce damage in the host tissues during EiWT challenge compared to Aquavac-ESC.

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