Purification method for the avian adeno-associated virus

Bauer, Hans-Joachim; Monreal, G

doi:10.1016/0166-0934(85)90127-2

Cited by 7 publications

(9 citation statements)

References 12 publications

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“…The helper activity depends on the presence of the herpesviruses (PRV, ILTV or HVT) and is not caused by contaminating avian adenovirus. This was concluded from the following observations (data not shown) : (i) chloroform treatment of the herpesvirus samples before co-infection with AAAV abolished the multiplication of infectious AAAV (the same chloroform treatment completely inactivates the herpesviruses used as helpers while avian adenovirus was found to be resistant to chloroform), and (ii) no contaminating avian adenovirus could be detected in our AAAV preparation by indirect immunofluorescence (Bauer & Monreal, 1985).…”

mentioning

confidence: 98%

“…The infectious AAAV particles which band in CsC1 at a density of 1.39 to 1.40 g/ml were collected. The remaining CELO virus in this AAAV preparation was removed by immunoprecipitation using rabbit anti-CELO virus serum and Protein A-Sepharose (Bauer & Monreal, 1985). Serum proteins in the purified AAAV were removed by an additional equilibrium density gradient.…”

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confidence: 99%

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Herpesviruses Provide Helper Functions for Avian Adeno-associated Parvovirus

Bauer

Monreal

1986

Journal of General Virology

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SUMMARYThe avian herpesviruses infectious laryngotracheitis virus (ILTV) and herpesvirus of turkeys (HVT), as well as the mammalian herpesvirus pseudorabies virus (PRV) were able to provide complete helper activity for the production of infectious avian adenoassociated virus (AAAV) in chicken cells. The presence of AAAV in the infected chicken cell reduced the multiplication of HVT. ILTV or PRV, however, were not affected if used as helper viruses. Infectious AAAV was determined by an indirect immunofluorescence assay and infectious herpesvirus by plaque assays.Avian adeno-associated virus (AAAV) is a defective parvovirus which grows efficiently only in chicken cells co-infected with avian adenovirus (Yates et al., 1973;Pronovost et al., 1979). This report investigates whether or not other avian viruses can provide the helper function required for active multiplication of AAAV in chicken cells. We demonstrate that two avian herpesviruses, infectious laryngotracheitis virus (ILTV) and herpesvirus of turkeys (HVT), as well as pseudorabies virus (PRV), a herpesvirus of mammalian origin, provide complete helper activity for AAAV. A helper activity of human herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) for human AAV has been demonstrated (Buller et al., 1981). We also show that the growth of HVT is reduced in chicken cells co-infected with HVT and AAAV. However, there is no inhibitory effect on the multiplication of the helper virus if PRV or ILTV is used as helper.The AAAV strain ATCC VR-865 was propagated in embryonated chicken eggs (VALO, Cuxhaven, F.R.G.) by co-infection with fowl adenovirus serotype l (CELO virus) and prepurified by 1,1,2-trichlorotrifluoroethane treatment, followed by centrifugation in one discontinuous and two CsC1 equilibrium density gradients. The infectious AAAV particles which band in CsC1 at a density of 1.39 to 1.40 g/ml were collected. The remaining CELO virus in this AAAV preparation was removed by immunoprecipitation using rabbit anti-CELO virus serum and Protein A-Sepharose (Bauer & Monreal, 1985). Serum proteins in the purified AAAV were removed by an additional equilibrium density gradient. The final virus suspension was dialysed against cell culture Medium 199 with Earle's salts.The non-pathogenic ILTV strain ASL (Gelenczei & Marty, 1964) was plaque-purified three times and propagated in primary chicken kidney cells (CKC). The FC 126 strain of HVT (Witter et al., 1970) was obtained as a live vaccine against Marek's disease from E. Vielitz (TAD, Cuxhaven, F.R.G.). The lyophilized virus material was dissolved in SPGA stabilizer, pH 6.5, containing 0.218 M-sucrose, 0'0038 M-monopotassium phosphate, 0.0072 M-dipotassium phosphate, 0.0049 M-monosodium glutamate and 1 ~ bovine serum albumin (Calnek et al., 1970). The DEK strain of PRV (kindly provided by H. Ludwig, Berlin, F.R.G.) was plaquepurified three times and propagated in secondary chicken embryo fibroblasts (CEF).To study the growth cycle of AAAV with ILTV as helper virus and the influence of AAAV on the multiplication ...

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confidence: 98%

mentioning

confidence: 99%

Herpesviruses Provide Helper Functions for Avian Adeno-associated Parvovirus

Bauer

Monreal

1986

Journal of General Virology

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“…was employed in all experiments. The virus was propagated in specific pathogen free (SPF) embryonating eggs (VALO) by coinfection with CELO virus (Bauer and Monreal, 1985). Three FAV strains were used in these studies: CELO phelps strain (FAV-1) from our laboratory, 340 strain (FAV-5) and Hungaria VI strain (FAV-8).…”

Section: Virus Strainsmentioning

confidence: 99%

“…The infectious AAAV particles which band in CsCl at a density of 1.39 to 1.40 g/cm 3 were collected. CELO virus still contaminating this AAAV population was removed by immunoprecipitation using rabbit anti CELO serum and protein A-Sepharose (Bauer and Monreal, 1985). Serum proteins in the purified AAAV were eliminated by an additional equilibrium density gradient.…”

Section: Purification Ofaaa Vmentioning

confidence: 99%

“…The rabbit anti CELO serum used for the immunoprecipitation of the CELO virus (Bauer and Monreal, 1985) was heat inactivated (56°C;30min), centrifuged at 25000 g for 30 min and dialysed against isotonic Tris buffer, pH 7.2 (Gambke and Deppert, 1983). For the indirect immunofluorescence test, the rabbit antisera against AAAV and CELO virus were not heat inactivated.…”

Section: Purification Ofaaa Vmentioning

confidence: 99%

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Avian adeno‐associated virus (AAAV) and fowl adenoviruses (FAV): Studies of viral interactions in chicken cell cultures

et al. 1986

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SUMMARYAs the growth kinetics of avian adeno-associated virus (AAAV) in chicken cells demonstrate, the three serotypes of fowl adenovirus (FAV), FAV -1, -5 and -8, provide complete helper activity for the production of infectious AAAV. Under one step conditions, the growth cycle of AAAV in primary chicken kidney cell (CKC) cultures is characterised by an eclipse phase of 8 hours and an exponential increase of the virus infectivity by 4 to 5 logs until 24 hours post-adsorption (p.a.). These growth characteristics do not depend on the serotype of FAV used as helper. In chicken embryo fibroblast (CEF) cultures the eclipse phase is prolonged to 12 hours p.a. and the virus infectivity increases only by 2 logs. In addition, the low efficiency of plating of FAV -1 in this cell system does not allow one step growth curves for AAAV. In CKC and CEF cultures coinfected with FAV and AAAV the multiplication of helper FAV is reduced. The degree of growth inhibition depends on the AAV multiplicity used. Sequential infection of CKC cultures with FAV -1 and AAAV modifies the AAAV growth cycle, i.e. there is a time reduction of the eclipse phase and a decrease of the virus yield. Infectious AAAV was determined by an indirect immunofluorescence assay and infectious FAV by a plaque assay.

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Biological and physicochemical characterization of the major (1.40) and minor (1.45) component of infectious avian adeno-associated virus

Bauer

Schneider

Gelderblom

et al. 1991

Archives of Virology

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Two infectious components with buoyant densities of 1.40 g/cm3 and 1.45 g/cm3, designated as major (1.40) and minor (1.45) component, were detected by banding avian adeno-associated virus (AAAV) isopycnically in CsCl. In metrizamide, however, infectious AAAV banded only as a single peak at a density of 1.32 g/cm3. Biological as well as physicochemical properties of the two AAAV components recovered from CsCl density gradient were described. Concerning the minor (1.45) component, three experimental findings may suggest that the capsid structure of this AAAV population is altered in comparison with that of the major (1.40) component: (i) the sedimentation pattern characterized by an additional peak containing slower-sedimenting noninfectious material (16 S); (ii) the specific infectivity decreased by the 3.5 fold; (iii) the ready disintegration when exposed to gently denaturing conditions.

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Purification method for the avian adeno-associated virus

Cited by 7 publications

References 12 publications

Herpesviruses Provide Helper Functions for Avian Adeno-associated Parvovirus

Herpesviruses Provide Helper Functions for Avian Adeno-associated Parvovirus

Avian adeno‐associated virus (AAAV) and fowl adenoviruses (FAV): Studies of viral interactions in chicken cell cultures

Biological and physicochemical characterization of the major (1.40) and minor (1.45) component of infectious avian adeno-associated virus

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