Transfection of monkey kidney cells with HAV cDNA or its RNA transcripts induced HAV infection, with RNA transcripts yielding about 10-fold more virus than DNA.
Infectious RNA transcripts from HAV cDNA can induce HAV infection in cell culture and animal models, providing a tool for studying viral functions and attenuation.
A full-length cDNA copy of an attenuated, cell culture-adapted hepatitis A virus (HAV HM-175/7 MK-5) genome was constructed in the PstI site of plasmid vector pBR322. Transfection of monkey kidney cells with this plasmid failed to induce the production of hepatitis A virus (HAV). The HAV cDNA was excised from pBR322 and inserted, without the oligo(dG) X oligo(dC) tails, into an RNA transcription vector to yield plasmid pHAV/7. Transfection of monkey kidney cells with pHAV/7 DNA induced HAV infection. Transfection with RNA transcripts produced in vitro from pHAV/7 yielded about 10-fold more HAV than did transfection with pHAV/7 DNA. Marmosets inoculated with transfection-derived virus developed anti-HAV antibodies and had liver enzyme patterns that closely resembled the liver enzyme patterns seen in animals inoculated with virus from a comparable level of cell culture passage. Infectious RNA transcripts from HAV cDNA should be useful for studying the molecular basis of cell culture adaptation and attenuation as well as for studying specific viral functions.
Cohen et al. (Thu,) conducted a other in Hepatitis A virus infection. Transfection with HAV cDNA (pHAV/7) and its RNA transcripts vs. Transfection with pBR322 plasmid / pHAV/7 DNA was evaluated on Production of infectious hepatitis A virus. Transfection of monkey kidney cells with HAV cDNA or its RNA transcripts induced HAV infection, with RNA transcripts yielding about 10-fold more virus than DNA.