[Isolation of adeno-associated virus type 4 from a culture of green monkey kidney cells]. / Vydelenie adenoassotsiirovannogo virusa tipa 4 iz kul'tury kletok pochek zelenykh martyshek.

In the process of biological control of uninfected green monkey kidney (GMK) cell cultures a thermostable hemagglutinating agent designated No. 5056 was isolated alongside with adenovirus-SV. By its antigenic properties the 5056 strain was identified as adeno-associated virus type 4 (AAV-4). In control of 574 specimens of GMK culture batches, 40 AAV-4 strains were isolated in the presence of a helper adenovirus. Some biological properties of the isolates and their resistance to certain physical and chemical treatments were studied. Two isolates of the satellite virus were examined in the electron microscope. A correlation between the rate of AAV-4 isolation from GMK cultures and the presence of complement-fixing antibody to AAV-4 in monkey sera was observed.

[Experimental infection of green monkeys with adenoassociated virus]. / Eksperimental'naia infektsiia zelenykh martyshek adenoassotsiirovannym virusom.

Primary infection and reinfection with adeno-associated virus type 4 (AAV-4) was reproduced in green monkeys experimentally infected with AAV-4 in mixture with adenovirus. Wide dissemination of the satellite virus in animals was observed. AAV-4 and its antigen were detectable 5 to 23 days after inoculation. In monkeys infected with a mixture of AAV-4 and adenovirus or with one of them the infection was accompanied by a marked fever persisting from the 5th to the 20th day after inoculation. The infected monkeys showed an intensive rise of homologous antibody titer most marked on the 10th-15th day after inoculation with AAV-4. AAV-4 and its antigen were detected in smears from conjunctival and tonsillar mucosa, rectal specimens in the time course of the infectious process, as well as from the trachea, lungs, liver, spleen, intestines and kidneys of the sacrificed monkeys. Besides, AAV-4 antigen was found in cells of the tonsils and blood leukocytes of the sacrificed monkeys. No virus or its antigen were found in the brain and heart tissues. Virions of adeno-associated virus were found by electron microscopic examinations of kidney cells of one of 3 monkeys infected with AAV-4.

[Helper viruses of adeno-associated virus type 4 replication]. / Virusy-pomoshchniki replikatsii adenoassotsiirovannogo virusa tipa 4.

In replication of adeno-associated virus type 4 (AAV-4) the helper function may be performed by a non-defective virus from the same group of parvoviruses (Kilham virus). The synthesis of AAV-4 antigen was observed in a pig embryo kidney cell line, SPEV, chronically infected with Kilham virus, strain RV-13, 45--52 passages. A one-day-old SPEV-Kilham culture was infected with AAV-4. The AAV-4 antigen was detected by immunofluorescence at 6, 8, 12, 18 hours, 2, 3, 4, and 5 days after inoculation. During the first 2--4 days after inoculation the AAV-4 antigen was found in the nucleus and perinuclear zone, later in the cytoplasm. A "new" helper virus for AAV-4 replication has been found: simiancytomegalovirus in human embryo fibroblast cell culture permissive for the helper virus. In the systems where AAV-4 replicates, its antigen can be detected in the nucleus and perinuclear zone by IF. AAV-4 did not replicate in a system insensitive to the helper virus or under non-permissive conditions: at the time, the AAV-4 antigen localized only in the cell cytoplasm was detected.