A dual role for adeno-associated virus in human health.

Adeno-associated virus (AAV) differs from most other viruses, as it requires the simultaneous presence of a helper virus for an active infection. Up to 80% of the human population is seropositive for AAV antibodies. AAV has been known to be a non-pathogenic virus and an inhibitor of carcinogenesis caused by coinfecting viruses. However, the recent reports associating AAV infection with hepatocellular carcinoma development and the mysterious cases of acute severe hepatitis in children have challenged the idea that AAV is a harmless virus. Herein, we explore the usefulness of AAV in gene therapy and the importance of AAV as a protector or perpetrator in human carcinogenesis, ultimately reflecting on the dual role of AAV in human health.

Adeno-associated virus and human papillomavirus types in cervical samples of pregnant and non-pregnant women.

OBJECTIVE: To investigate the prevalence of AAV and HPV DNA and their types in cervical secretion from pregnant and non-pregnant women. STUDY DESIGN: The samples were obtained from 40 pregnant and 62 non-pregnant women who were attended at the outpatient clinic of the Federal University Hospital of Espírito Santo, Southeastern Brazil. AAV and HPV were investigated by PCR and typed by PCR and/or RFLP. RESULTS: The occurrence of AAV in all samples was 25.5% (26/102): 81% (21/26) and 19% (5/26) for AAV2/3 and AAV5 species, respectively. AAV were observed in 35% (14/40) and 19% (12/62) of pregnant and non-pregnant women, respectively. HPV occurred in 22% of all samples; 25% (10/40) in pregnant and 20% (12/60) in non-pregnant women. HPV types were determined for 72.7% of the strains, most of which classified as high-risk. AAV-HPV co-infection was observed in 15.4% (4/26), mostly in pregnant women. CONCLUSIONS: There was a greater prevalence of AAV and HPV in pregnant than in non-pregnant women, which suggests that the gestational state may play a role in reactivating the viruses.

Replicon-helper systems from attenuated Venezuelan equine encephalitis virus: expression of heterologous genes in vitro and immunization against heterologous pathogens in vivo.

A replicon vaccine vector system was developed from an attenuated strain of Venezuelan equine encephalitis virus (VEE). The replicon RNA consists of the cis-acting 5' and 3' ends of the VEE genome, the complete nonstructural protein gene region, and the subgenomic 26S promoter. The genes encoding the VEE structural proteins were replaced with the influenza virus hemagglutinin (HA) or the Lassa virus nucleocapsid (N) gene, and upon transfection into eukaryotic cells by electroporation, these replicon RNAs directed the efficient, high-level synthesis of the HA or N proteins. For packaging of replicon RNAs into VEE replicon particles (VRP), the VEE capsid and glycoproteins were supplied in trans by expression from helper RNA(s) coelectroporated with the replicon. A number of different helper constructs, expressing the VEE structural proteins from a single or two separate helper RNAs, were derived from attenuated VEE strains Regeneration of infectious virus was not detected when replicons were packaged using a bipartite helper system encoding the VEE capsid protein and glycoproteins on two separate RNAs. Subcutaneous immunization of BALB/c mice with VRP expressing the influenza HA or Lassa virus N gene (HA-VRP or N-VRP, respectively) induced antibody responses to the expressed protein. After two inoculations of HA-VRP, complete protection against intranasal challenge with influenza was observed. Furthermore, sequential immunization of mice with two inoculations of N-VRP prior to two inoculations of HA-VRP induced an immune response to both HA and N equivalent to immunization with either VRP construct alone. Protection against influenza challenge was unaffected by previous N-VRP immunization. Therefore, the VEE replicon system was characterized by high-level expression of heterologous genes in cultured cells, little or no regeneration of plaque-forming virus particles, the capability for sequential immunization to multiple pathogens in the same host, and induction of protective immunity against a mucosal pathogen.

Evolutionary relationship between plasmids and phages: phasmid P4 as a model.

Phasmid (phage-plasmid) P4 is capable of propagating as a temperate phage, lytic helper-dependent phage, and as a plasmid (pP4vir1). Alteration of cellular growth conditions can be used to manipulate the copy number (n) of pP4vir1 from low (n = 1-2) to high (n = 30-40). The levels of gene expression of P4 in the plasmid and phage states are radically different. While there is high expression of P4 proteins in the phage state, low levels of gene expression are found in the plasmid mode of propagation thereby allowing pP4vir1 to remain intracellularly without perturbing host homeostasis. Possible evolutionary origins of phasmid P4 are discussed.