Functional gene transfer from intracellular bacteria to mammalian cells

We provide evidence of direct transfer of functional DNA from bacteria to mammalian cells. An Escherichia coli K12 diaminopimelate auxotroph made invasive by cloning the invasin gene from Yersinia pseudotuberculosis transfers DNA after simple co-incubation, into a variety of mammalian cell lines. Transfer efficiency was enhanced in some cells by coexpression of the gene for listeriolysin from Listeria monocytogenes. Expression of the acquired genes occurs in both dividing and quiescent cells. The only requirement for bacteria to transfer genetic material into nonprofessional phagocytic cells and macrophages is the ability to invade the host cell.

[1]  R. Steinman,et al.  The endocytic activity of dendritic cells , 1995, The Journal of experimental medicine.

[2]  Pascale Fung,et al.  Listeriolysin O is essential for virulence of Listeria monocytogenes: direct evidence obtained by gene complementation , 1989, Infection and immunity.

[3]  E. Chi,et al.  Group A streptococci efficiently invade human respiratory epithelial cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[4]  J. Sadoff,et al.  Attenuated bacteria as a DNA delivery vehicle for DNA-mediated immunization. , 1997, Vaccine.

[5]  V. Buchanan-Wollaston,et al.  The mob and oriT mobilization functions of a bacterial plasmid promote its transfer to plants , 1987, Nature.

[6]  S. Falkow,et al.  Identification of invasin: A protein that allows enteric bacteria to penetrate cultured mammalian cells , 1987, Cell.

[7]  W. Goebel,et al.  Delivery of antigen-encoding plasmid DNA into the cytosol of macrophages by attenuated suicide Listeria monocytogenes , 1998, Nature Biotechnology.

[8]  E. Chi,et al.  Invasion of respiratory epithelial cells by Burkholderia (Pseudomonas) cepacia , 1996, Infection and immunity.

[9]  D. Belin,et al.  A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors. , 1984, Gene.

[10]  P. Trieu-Cuot,et al.  Plasmid transfer by conjugation from Escherichia coli to Gram‐positive bacteria , 1987 .

[11]  J. Thomas August,et al.  Purification and characterization of human lysosomal membrane glycoproteins. , 1989, Archives of biochemistry and biophysics.

[12]  D. Heitmann,et al.  Frequency and conditions of spontaneous plasmid transfer from E. coli to cultured mammalian cells. , 1993, Bio Systems.

[13]  K. Kelly,et al.  Internalization of Chlamydia by dendritic cells and stimulation of Chlamydia-specific T cells. , 1998, Journal of immunology.

[14]  S. Falkow,et al.  A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12 , 1985, Nature.

[15]  P. Courvalin,et al.  Gene transfer from bacteria to mammalian cells. , 1995, Comptes rendus de l'Academie des sciences. Serie III, Sciences de la vie.

[16]  P. Youngman,et al.  Bacillus subtilis expressing a haemolysin gene from Listeria monocytogenes can grow in mammalian cells , 1990, Nature.

[17]  P. Sansonetti,et al.  Involvement of a plasmid in the invasive ability of Shigella flexneri , 1982, Infection and immunity.

[18]  W. Schaffner.  Direct transfer of cloned genes from bacteria to mammalian cells. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[19]  J. Heinemann,et al.  Bacterial conjugative plasmids mobilize DNA transfer between bacteria and yeast , 1989, Nature.

[20]  K. Timmis,et al.  Oral Somatic Transgene Vaccination Using Attenuated S. typhimurium , 1997, Cell.

[21]  M. Kovacsovics-Bankowski,et al.  A phagosome-to-cytosol pathway for exogenous antigens presented on MHC class I molecules , 1995, Science.

[22]  R. Isberg,et al.  Multiple β 1 chain integrins are receptors for invasin, a protein that promotes bacterial penetration into mammalian cells , 1990, Cell.

[23]  P. Ricciardi-Castagnoli,et al.  Immortalized dendritic cell line fully competent in antigen presentation initiates primary T cell responses in vivo , 1993, The Journal of experimental medicine.

[24]  J. Sadoff,et al.  Attenuated Shigella as a DNA Delivery Vehicle for DNA-Mediated Immunization , 1995, Science.

[25]  S. Falkow,et al.  Analysis of expression and thermoregulation of the Yersinia pseudotuberculosis inv gene with hybrid proteins , 1988, Infection and immunity.

[26]  P. Sansonetti,et al.  Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri , 1983, Infection and immunity.