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The development of the real-time RT-PCR represents a major advance in quantitation of JSRV. Although it does not give an estimate of infectivity, the assay will allow the matching of JSRV dose in different experimental inocula and also in future vaccine components.
Antisera was raised in rabbits against different recombinant JSRV proteins during the course of this project. The antisera has already been supplied to a number of labs around the world for various applications in research studies e.g. immunohistochemistry, Western blotting. This includes research on JSRV/OPA and also on human cancers.
Antibodies were also raised in sheep against recombinant JSRV CA. The result confirms that sheep are not inherently immunotolerant to JSRV, despite the presence of closely releted endogenous retroviruses in the sheep genome. Equivalent antibody responses were induced in uninfected and in JSRV-infected sheep showing that infection with JSRV does not specifically block production of antibody to the virus. This is important information for any potential vaccine and shows that a post-infection vaccine may also be successful if it can prevent JSRV infection progressing to development of OPA.
A number of different new plasmid constructs were made for the expression of recombinant JSRV proteins. Expression and purification of the proteins was optimised and the proteins were used as immunogens or as antigens for immunological assays. The proteins have further use for similar studies. In the right dose, adjuvant and immunisation route one or more of the proteins may be a component of a future vaccine against OPA. The same proteins could also be employed in assays to monitor vaccine efficacy in inducing immune responses.
The assays to measure immune responses against JSRV have future utility in various aspects of OPA research. They are necessary and appropriate for use in continued studies aimed at vaccine development. They are also useful for basic research into the pathogenesis of OPA.
The generation of full length clones of caENTV has several applications in research and potentially in vaccine development. Sequence data acquired can be used in comparative studies. Clones can be used in expression studies aimed at delineating the pathogenesis of caENTV and the related viruses JSRV and ovENTV. Infectious molecular clones can be used to produce caENTV in vitro as a defined source of the virus for challenge studies.
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