THE GLYCOPROTEINS OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS AND THEIR ROLE IN INFECTION AND IMMUNITY

The porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important pathogen of swine and is known to cause abortion and infertility in pregnant sows and respiratory distress in piglets. PRRSV contains a major glycoprotein (GP5) and three minor glycoproteins (GP2a, GP3, and GP4) on the virion envelope, all of which are required for infectious virus production. To study their interactions amongst each other and with a cellular receptor for PRRSV, CD163, I cloned each of the viral glycoproteins and CD163 in various expression vectors. My studies have shown that while the GP2a, GP3, and GP4 are co-translationally glycosylated, the GP5 is post-translationally glycosylated. By using co-immunoprecipitation (co-IP) assays, strong interaction was demonstrated between GP4 and GP5 proteins, although weak interactions among the other envelope glycoproteins were also detected. Further, GP4 was found to mediate interactions leading to formation of multiprotein glycoprotein complex. My results also show that GP2a and GP4 proteins are the only two GPs that specifically interact with the CD163 molecule and that glycosylation of these GPs is required for efficient interaction. Based on these studies, I have developed an interactome map of the viral GPs and CD163 and have proposed a model of the viral glycoprotein complex and its interaction with CD163. Studies reported here also show that glycan addition at residue 184 (N184) of GP2a, and residues N42, N50, and N131 of GP3 is essential for recovery of infectious virus. Although single site glycosylation mutants of GP4 had no effect on infectious virus production, introduction of double mutations was lethal. The loss of glycan moieties of GP2a, GP3, and GP4 proteins had no effect on host neutralizing antibody production. Overall, I conclude that the PRRSV glycoproteins are co-translationally and post-translationally glycosylated, the GP4 protein is central to mediating interglycoprotein interactions, and along with GP2a, serves as the viral attachment protein that is responsible for interactions with the viral receptor, CD163. Further, glycosylation of GP2a, GP3, and GP4 proteins is required for infectious virus production, efficient interaction with CD163, but does not play any role in neutralizing antibody response in infected animals.

The Perception of Natural, Cell Phone, and Computer-Synthesized Speech During The Performance Of Simultaneous Visual-Motor Tasks

This study investigated the influence of top-down and bottom-up information on speech perception in complex listening environments. Specifically, the effects of listening to different types of processed speech were examined on intelligibility and on simultaneous visual-motor performance. The goal was to extend the generalizability of results in speech perception to environments outside of the laboratory. The effect of bottom-up information was evaluated with natural, cell phone and synthetic speech. The effect of simultaneous tasks was evaluated with concurrent visual-motor and memory tasks. Earlier works on the perception of speech during simultaneous visual-motor tasks have shown inconsistent results (Choi, 2004; Strayer & Johnston, 2001). In the present experiments, two dual-task paradigms were constructed in order to mimic non-laboratory listening environments. In the first two experiments, an auditory word repetition task was the primary task and a visual-motor task was the secondary task. Participants were presented with different kinds of speech in a background of multi-speaker babble and were asked to repeat the last word of every sentence while doing the simultaneous tracking task. Word accuracy and visual-motor task performance were measured. Taken together, the results of Experiments 1 and 2 showed that the intelligibility of natural speech was better than synthetic speech and that synthetic speech was better perceived than cell phone speech. The visual-motor methodology was found to demonstrate independent and supplemental information and provided a better understanding of the entire speech perception process. Experiment 3 was conducted to determine whether the automaticity of the tasks (Schneider & Shiffrin, 1977) helped to explain the results of the first two experiments. It was found that cell phone speech allowed better simultaneous pursuit rotor performance only at low intelligibility levels when participants ignored the listening task. Also, simultaneous task performance improved dramatically for natural speech when intelligibility was good. Overall, it could be concluded that knowledge of intelligibility alone is insufficient to characterize processing of different speech sources. Additional measures such as attentional demands and performance of simultaneous tasks were also important in characterizing the perception of different kinds of speech in complex listening environments.