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.

Contesting Sphere Boundaries Online: Private/Technical/Public Discourses in Polycystic Ovarian Syndrome Discussion Groups

The internet is fast becoming a means for people to obtain information, creating a unique forum for the intersection of the public, technical, and private spheres. To ground my research theoretically, I used Jürgen Habermas’s sphere theory. Habermas (1987) explains that the technical sphere colonizes the private sphere, which decreases democratic potential. In particular, the internet is a place for altering technical colonization of the private and public spheres. My research focuses on women’s health because it is a particularly useful case study for examining sphere tensions. Historically, the biomedical health establishment has been a powerful agent of colonization, resulting in detrimental effects for women and their health. The purpose of this study is to examine how the internet encourages expert and female patient deliberation, which empowers women to challenge the experts and, thus, make conversations between the private/technical spheres more democratic. I used PCOS (Polycystic Ovarian Syndrome) as a case to observe the changing sphere boundaries by studying the discourse that took place on multiple patient and doctor websites over a four-year period. Through my research, I found that the PCOS women challenge the biomedical model by appropriating medical language. By understanding the medical talk, the women are able to feel confident when discussing their health conditions with the doctor and with each other. The PCOS women also become lay-experts who have personal and medical experience with PCOS, reducing private sphere colonization. This case study exemplifies how female empowerment can influence expert culture, challenging our conventional understanding of democracy.