Culturing circular ssDNA viruses for the production of vaccines
AssigneeUniversiteit Gent
InventorsHans Nauwynck, Gerald Misinzo, Sven Arnouts
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Abstract
The present invention relates to the use of interferon in the in vitro cultivation of animal circular ssDNA virus such as Porcine Circovirus 2 or human TT virus in an animal cell line. Increased titres of animal circular ssDNA virus are obtained by addition of interferons or agents which ensure the production of endogenous interferons by said cell line and/or by the reduction of endosomal-lysosomal system acidification.
Claims35
A method for the in vitro cultivation of an animal circular ssDNA virus comprising the step of (a) inoculating cells of a continuous animal cell linein a culture medium with said circular ssDNA virus, (b) cultivating said continuous animal cell line in said medium, thereby ensuring that endosomal-lysosomal system acidification is reduced in said continuous animal cell line, and (c) isolating said animal circular ssDNA virus from said medium and/or said infected continuous animal cell line.
An undiluted cultivation medium of an in vitro culture of a cell-line comprising animal circular ssDNA virus characterised in that it further comprises an agent capable of inhibiting endosomal-lysosomal system acidification.
An undiluted cultivation medium of an in vitro culture of a cell-line comprising animal circular ssDNA virus characterised in that it further comprises at least 2 U/ml interferon.
A method for producing a vaccine for protection against an animal circular DNA virus, comprising said animal circular DNA virus or components thereof, said method comprising the steps of:
- a) inoculating cells of a continuous animal cell line in a culture medium with said circular ssDNA virus, thereby ensuring, prior to and/or after said inoculation, the reduction of endosomal-lysosomal system acidification of said continuous animal cell line;
- b) allowing said ssDNA virus to replicate in said continuous animal cell line;
- and
- c) obtaining said circular ssDNA virus or components thereof from said continuous animal cell line or said medium.
An in vitro method for determining the infection of a sample by an animal circular ssDNA virus comprising increasing the amount of virus before detection using endosomal-lysosomal system acidification inhibitors.
A method of improving the immune response of a subject to a vaccine comprising an attenuated animal circular ssDNA virus, said method comprising, administering one or more inhibitors of endosomal-lysosomal system acidification to said subject, simultaneously with or shortly before or after administering said vaccine to said subject.
A vaccine comprising an attenuated animal circular ssDNA virus and one or more inhibitors of endosomal-lysosomal system acidification.
Dependencies are inferred from the claim text.