Absence of germline infection in male mice following intraventricular injection of adenovirus.

Document Type

Article

Publication Date

2001

Keywords

Animals, Cerebral-Ventricles, DNA-Primers, Fluorescent-Antibody-Technique-Indirect, Gene-Therapy, Gene-Transfer-Techniques, Humans, Injections-Intraventricular, Lac-Operon, Male, Membrane-Proteins, Mice, Polymerase-Chain-Reaction, Receptors-Virus, Spermatozoa, Testis, beta-Galactosidase

First Page

603

Last Page

613

JAX Source

Mol Ther 2001 Dec; 4(6):603-13.

Abstract

The possibility of inadvertent exposure of gonadal tissue to gene therapy vectors has raised safety concerns about germline infection. We show here that the receptor for coxsackie B viruses and adenoviruses 2 and 5 (CXADR) is expressed in mouse germ cells, suggesting the possibility that these viruses could infect germ cells. To directly assess the risk of germline infection in vivo, we injected an adenovirus carrying the germ-cell-specific protamine promoter fused to the bacterial lacZ reporter gene into the left ventricular cavity of mice and then monitored expression of the reporter gene in germ cells. To differentiate between infection of stem cells and differentiating spermatogenic cells, we analyzed expression of the reporter cassette at different times after viral delivery. Under all conditions tested, mice did not express the Escherichia coli beta-galactosidase protein in developing spermatids or in mature epididymal spermatozoa. Primary germ cells cultured in vitro were also refractory to adenoviral infection. Our data suggest that the chance of vertical germline transmission and insertional mutagenesis is highly unlikely following intracoronary adenoviral delivery.

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