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Studies of the biological function of avian reovirus proteins

Astawa, I Nyoman Mantik (1995) Studies of the biological function of avian reovirus proteins. PhD thesis, Murdoch University.

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Abstract

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Monoclonal antibodies (MAbs) against virion proteins of avian reovirus strain 1091 were produced and used to study the structural and biological function of avian reovirus proteins.

Seventeen anti-reovirus MAbs of immunoglobulin G (IgG) isotype were produced. Of 10 virus-specific proteins (with molecular weights of 135,120, 105, 76, 73, 71,46,43,41, and 38 KDa and designated as A, A.B, C, UA, UB, UBC, oA, oB, oNS and oC, respectively)* demonstrated by radioimmunoprecipitation (RIP) and/or Western blotting with polyclonal antiserum, the MAbs reacted with B, a complex of uB and uBC, oA, oNS, and oC.

Some MAbs reactive with uB or the uB/uBC complex were also reactive with oB. The results suggested that uBC was a cleavage product of uB and that the uB/uBC proteins have a strong affinity with oB.

Plaque reduction neutralisation assay using anti-1091 MAbs revealed that three proteins - B, uB/uBC and oC - were involved in virus neutralisation in Vero cells but only two proteins - uB/uBC and oC - were involved in vims neutralisation in chicken kidney (CK) cells. One MAb against the pB/pC proteins and some MAbs against the oC protein neutralised the vims to a very high titre suggesting that these two proteins were associated with the virus-cell interaction during the initial stages of infection process. An additional protein A,B was also associated with vims neutralisation but probably by a steric hindrance of uB/uBC and oC as one MAb against this protein neutralised the vims to a very low titre.

An important biological characteristic of cells infected with avian reovims is vims-induced cell-fusion. During studies of the effect of MAbs against virion proteins of avian reovimses 1091 and RAMI, two probably cell fusion-related phenomena were observed: inhibition of cell fusion in infected cells overlayed with liquid medium, and inhibition of plaque development in infected cells overlayed with agar containing medium. At least two proteins (the pB/pBC complex, and oC) were involved in avian reovirus-induced cell fusion: MAbs against these two proteins inhibited both vims-induced cell fusion and plaque development, and both the uB/uBC and oC proteins were detected on the surface of infected cells by RIP and immunofluorescence using both MAbs and polyclonal antiserum. Differences between the activity of different (uB/uBC and oC MAbs suggested that although these two phenomena were related events, they were associated with different epitopes on the viral proteins.

By indirect immunofluorescence and neutralisation tests it was determined that MAbs against oC of avian reovirus 1091 cross-reacted only with oC of viruses of the homologous antigenic type, confirming previous reports that oC was the type-specific avian reovirus protein. Anti oC MAbs neutralised virus infectivity in both Vero and CK cells. The XB protein was associated with group (broad)-specific virus neutralisation in Vero cells and not in CK cells. The uB/uBC complex was also associated with group (broad) specific virus neutralisation but in both Vero and in CK cells.

Item Type: Thesis (PhD)
Murdoch Affiliation: School of Veterinary Studies
Notes: Note to the author: If you would like to make your thesis openly available on Murdoch University Library's Research Repository, please contact: repository@murdoch.edu.au. Thank you.
Supervisor(s): Wilcox, Graham
URI: http://researchrepository.murdoch.edu.au/id/eprint/51793
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