Pseudomonas exotoxin antisense RNA selectively kills hepatitis B virus infected cells.
OBJECTIVE
To present an approach for selectively killing retrovirus-infected cells that combines the toxicity of Pseudomonas exotoxin (PE) and the presence of reverse transcriptase (RT) in infected cells.


METHODS
PE antisense toxin RNA has palindromic stem loops at its 5' and 3' ends enabling self-primed generation of cDNA in the presence of RT. The RT activity expressed in retrovirus-infected cells converts "antisense-toxin-RNA" into a lethal toxin gene exclusively in these cells.


RESULTS
Using cotransfection studies with PE-expressing RNAs and beta-gal expressing reporter plasmids, we show that, in HepG2 and HepG2.2.15 hepatoma cells as well as in duck hepatitis B virus (DHBV) infected cells, HBV or DHBV-polymerase reverse transcribe a lethal cDNA copy of an antisense toxin RNA, which is composed of sequences complementary to a PE gene and eukaryotic transcription and translation signals.


CONCLUSIONS
This finding may have important implications as a novel therapeutic strategy aimed at the elimination of HBV infection.
Hafkemeyer
Peter
P
Department of Medicine II, University Hospital Freiburg, Hugstetterstrasse 55, Freiburg D-79106, Germany. phafkemeyer@gmx.net
Brinkmann
Ulrich
U
Brinkmann
Elizabeth
E
Pastan
Ira
I
Blum
Hubert-E
HE
Baumert
Thomas-F
TF
D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000821:virology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004279:DNA, Viral / Q000235:genetics; D005098:Exotoxins / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D006509:Hepatitis B / Q000628:therapy; D006515:Hepatitis B virus / Q000187:drug effects / Q000502:physiology*; D004371:Hepatitis Virus, Duck / Q000187:drug effects / Q000502:physiology; D018197:Hepatoblastoma / Q000473:pathology / Q000821:virology*; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology / Q000821:virology*; D010957:Plasmids / Q000235:genetics; D011549:Pseudomonas / Q000378:metabolism*; D016372:RNA, Antisense / Q000235:genetics / Q000494:pharmacology*; D048348:Reverse Transcription / Q000235:genetics; D014162:Transfection
D004279:DNA, Viral; D005098:Exotoxins; D016372:RNA, Antisense