Characterization of Heavy Particles of Adeno-associated Virus Type 1

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Abstract
The temperature-sensitive mutant ts4 of adenovirus type 2 (Ad-2) is capable of complementing adeno-associated virus type 1 (AAV-1) in human pharyngeal carcinoma HEp2, oral carcinoma KB and embryonic kidney HEK cells at 34.degree. C and 39.degree. C when used as a helper virus. Heavy non-infectious AAV-1 particles can be generated by using the mutant ts4 in HEp2. When AAV-1 is grown in serial passages in HEp2 cells, both the wild-type Ad-2 and the mutant ts4 give rise to heavy, less infectious AAV-1 particles. The heavy AAV-1 particles generated by Ad-2 in advanced serial passages retain the property of having complement-fixing and immunofluorescent antigens, but the AAV-1 generated by the mutant in advanced serial passages lose this property. There is no appreciable difference in the particle counts made by EM of AAV-1 preparations generated either by Ad-2 or the mutant ts4. Analysis by polyacrylamide gel electrophoresis of purified heavy AAV generated by ts4 indicates that in late passage an additional polypeptide of higher MW than the 3 structural polypeptides is detected.