Wild-type intracellular bacteria deliver DNA into mammalian cells.
Gene transfer in vitro from intracellular bacteria to mammalian phagocytic and non-phagocytic cells and in vivo in mice has been reported. The bacteria used as DNA delivery vectors were engineered to lyze upon entry in the cell due to impaired cell wall synthesis for Shigella flexneri and invasive Escherichia coli, or production of a phage lysin for Listeria mono- cytogenes. In vivo gene transfer was obtained with attenuated Salmonella typhimurium and resulted in stimulation of mucosal immunity. We report that wild-type intracellular human pathogens, such as L. monocytogenes EGD or LO28 and S. flexneri M90T, mediate efficient in vitro transfer of functional genes into epithelial and macrophage cell lines. A low- efficiency transfer was obtained from strain EGD to mouse peritoneal macrophages. DNA transfer with S. typhimurium was observed only from atten-uated aroA strain SL7207 into COS-1 cell line. As demonstrated by the study of listeriolysin-defective L. monocytogenes or of S. typhimurium SL7207 aroA engineered to secrete listeriolysin, escape of bacteria or of plasmid DNA from the intracytoplasmic vacuole is required for transfer of genetic information to occur.
Grillot-Courvalin
Catherine
C
Unité des Agents Antibactériens, Institut Pasteur, 75724 Paris Cedex 15, France. ccourval@pasteur.fr
Goussard
Sylvie
S
Courvalin
Patrice
P
D000818:Animals; D002460:Cell Line; D004269:DNA, Bacterial / Q000235:genetics*; D004755:Enterobacteriaceae / Q000235:genetics* / Q000472:pathogenicity; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008089:Listeria monocytogenes / Q000235:genetics* / Q000472:pathogenicity; D017737:Macrophages, Peritoneal / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics*
D004269:DNA, Bacterial