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.

Health Assessment of Weddell Seals, Leptonychotes weddellii, in McMurdo Sound, Antarctica

The demography of Weddell seals in eastern McMurdo Sound, Antarctica, has been well studied during the past three decades (e.g. Stirling 1971; Siniff et al. 1977; Testa and Siniff 1987; Hastings and Testa 1998; Gelatt et al. 2001). Detailed life-history data are available on thousands of seals tagged as pups in McMurdo Sound, making this population a rich resource for wildlife health studies because health parameters can be evaluated in the light of reproductive histories and genetic relationships of several generations of tagged seals. Recently, evidence of exposure to diseases generally associated with domestic animals and feral wildlife has been detected in Antarctic wildlife (Austin and Webster 1993; Olsen et al. 1996; Gardner et al. 1997; Retamal et al. 2000; Foster et al. 2002) and this has generated concern and debate regarding the risks of disease introduction to Antarctic wildlife. Antibodies to viruses that have caused large die-offs in phocids in other areas of the world have been detected in Weddell seals (Bengtson et al. 1991), and there is a historical report of a mass die-off of crabeater seals that may have had a viral etiology (Laws and Taylor 1957).