Heather D. Mayor

Antibodies to adeno-associated satellite virus and herpes simplex in sera from cancer patients and normal adults

The ecologic aspects of the distribution of adeno-associated satellite virus (ASV) in the human population are of great interest because of its unconditional defectiveness and dependence on adenovirus for full and herpesvirus for partial complementation. Adenoviruses and herpesviruses are extremely common and persistent infections in man. We have developed immunofluorescent procedures for detecting the presence of satellite virus antibodies in human sera. The percentage of sera with antibodies to the ASV 2-3 complex was significantly higher in the normal group than in the cancer patients whereas there were no significant differences in herpes antibodies between the groups. The low incidence of satellite antibodies was particularly striking in patients with genital malignancies. The role of ASVs in human disease is not known. Their role in possible abrogation of oncogenesis mediated through adenoviruses or herpesviruses is worthy of further investigation.

Plus and Minus Single-Stranded DNA Separately Encapsidated in Adeno-Associated Satellite Virions

Based on physical and chemical determinations, the mnolecular weight of the type 4 adeno-satellite virus is 5.4 X 106 daltons, and the virion contains 1.4 X 106 daltons of DNA. Denaturation and renaturation studies indicate that the viral genome is a single-stranded DNA molecule and that each virion contains either a minus or a plus strand. Upon extraction, the minus and plus strands unite to form double-stranded DNA molecules with no obvious excess of unpaired strands.

Nonencapsidated Infectious DNA of Adeno-Satellite Virus in Cells Coinfected with Herpesvirus

Adeno-associated satellite viruses produce antigen detectable by immunofluorescence but not infectious virus in tissue culture cells coinfected with herpes simplex virus. Analysis of DNA extracts from these infected cells shows that large amounts of infectious satellite virus DNA are produced but not encapsidated in the system. This result indicates that satellite virus may be defective at the maturation step.

Structural polypeptides of parvoviruses.

Abstract The structural polypeptides of X14 virus and adenovirus-associated viruses (AAV) types 1 and 4 were analyzed by SDS-polyacrylamide gel electrophoresis. The capsid of each defective parvovirus consists of two minor and one major polypeptide, but the capsid of X14 virus consists of only two polypeptides. The smallest polypeptide of each virus is the most abundant. The naturally occurring empty capsids of AAV type 4 and X14 have proteins indistinguishable in size and concentration from those of complete virions. The isolated structural polypeptides of the two hemagglutinating viruses, AAV4 and X14, did not show any hemagglutinating activity.

Transplacental Infection with Adeno-Associated Virus Type 1 in Mice

Adeno-associated type 1 parvovirus (AAV) was detected in the kidneys and lungs of fetuses and newborns, when pregnant mice were injected subcutaneously with AAV type 1 and murine adenovirus as a helper virus. These findings clearly indicate that transplacental infection with AAV in rodents has been achieved.

Isoelectric Focusing of Parvoviruses

Adenovirus-associated virus type 4 and X14 migrate during electrophoresis to pH 2.6 in sucrose-stabilized, pH 2.5-6.0 gradients. Naturally occurring empty capsids appear to have the same isoelectric point as complete virus particles.

Complementation of adeno-associated satellite viral antigens and infectious DNA by temperature-sensitive mutants of herpes simplex virus

Abstract Production of type 4 adeno-associated satellite virus (ASV) structural antigens and infectious DNA was studied using temperature-sensitive ( ts ) mutants of herpes simplex virus (HSV) types 1 and 2. Seven of the eight mutants studied were unable to synthesize HSV DNA at the nonpermissive temperature. Mutants belonging to three HSV-1 and two HSV-2 complementation groups complemented ASV antigen production while members of two other HSV-1 groups did not. Infectious satellite virus DNA was isolated from the HSV-1 ts -1 complementing system.

Complementation of Adeno-Associated Satellite Virus (AAV) by Temperature-Sensitive Mutants of Adenovirus Type 31

Temperature-sensitive (ts) matants of human adenovirus type 31 were able to complement adeno-associated satellite virus (AAV) antigen production in both HEK and KB cells at both permissive and non-permissive temperatures. However, mutant ts 94, an adenovirus 31 mutant which produces apparently normal amounts of structural protein and DNA but is defective in maturation, was significantly inhibited in its ability to potentiate AAV infectivity at the non-permissive temperature. Normal AAV DNA and adenovirus DNA were isolated from co-infections with AAV and mutant ts 94 at the non-permissive temperature. We suggest that an adenovirus-coded maturation function common to both adenovirus and AAV maturation is defective in the ts 94 system.

Adenovirus-associated virus polypeptides synthesized in cells coinfected with either adenovirus or herpesvirus

Abstract One major and two minor structural polypeptides of adenovirus-associated virus (AAV) were synthesized in AAV-infected cells coinfected with either adenovirus or herpesvirus as helper. The molar proportions of these polypeptides appeared to be the same as those found in the virion. Transport of these polypeptides from the cytoplasm to the nucleus occurred rapidly. No evidence was found for the synthesis of a large precursor protein in either system. Since complete AAV virions are not found in AAV-infected cells coinfected with herpesvirus, these findings indicate a failure in virus assembly in this system. This failure may be expressed at the level of DNA strand segregation or encapsidation of DNA, or in faulty capsid assembly.

Identification of a region of the HSV-1 genome with helper activity for AAV

Abstract A specific fragment of the herpes simplex virus type I genome has been shown to have the ability to provide helper activity for adeno-associated virus replication. Restriction endonuclease generated DNA fragments from either herpes viral DNA or recombinant plasmids were transfected into cultured cells. The cells were then infected with adeno-associated virus, and helper activity was detected by fluorescent antibody staining of the cells for adeno-associated virus infected antigens. The region with AAV helper activity is 1.75 kb in size and makes up only about 1% of the HSV-1 genome. It is located between map coordinates 0.316 and 0.327 on the prototype arrangement of the HSV-1 genome and is the first function to be mapped in this region.