This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Mitev, G. M.
Right arrow Articles by Geller, B. L.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mitev, G. M.
Right arrow Articles by Geller, B. L.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, September 2009, p. 3700-3704, Vol. 53, No. 9
0066-4804/09/$08.00+0     doi:10.1128/AAC.00099-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Inhibition of Intracellular Growth of Salmonella enterica Serovar Typhimurium in Tissue Culture by Antisense Peptide-Phosphorodiamidate Morpholino Oligomer {triangledown}

Georgi M. Mitev,1 Brett L. Mellbye,2,{dagger} Patrick L. Iversen,2 and Bruce L. Geller1,2*

Oregon State University,1 AVI BioPharma, Inc., Corvallis, Oregon2

Received 22 January 2009/ Returned for modification 2 March 2009/ Accepted 19 May 2009

Two types of phosphorodiamidate morpholino oligomers (PMOs) were tested for inhibition of growth of Salmonella enterica serovar Typhimurium. Both PMOs have the same 11-base sequence that is antisense to the region near the start codon of acpP, which is essential for lipid biosynthesis and viability. To the 3' end of each is attached the membrane-penetrating peptide (RXR)4XB (R, X, and B indicate arginine, 6-aminohexanoic acid, and β-alanine, respectively). One peptide-PMO (AcpP PPMO) has no charge on the PMO moiety. The second PPMO has three cations (piperazine) attached to the phosphorodiamidate linkages (3+Pip-AcpP PPMO). A scrambled-sequence PPMO (Scr PPMO) was synthesized for each type of PMO. The MICs of AcpP PPMO, 3+Pip-AcpP PPMO, and either one of the Scr PPMOs were 1.25 µM (7 µg/ml), 0.156 µM (0.94 µg/ml), and >160 µM (>900 µg/ml), respectively. 3+Pip-AcpP PPMO at 1.25 or 2.5 µM significantly reduced the growth rates of pure cultures, whereas AcpP PPMO or either Scr PPMO had no effect. However, the viable cell count was significantly reduced at either concentration of 3+Pip-AcpP PPMO or AcpP PPMO, but not with either Scr PPMO. In other experiments, macrophages were infected intracellularly with S. enterica and treated with 3 µM 3+Pip-AcpP PPMO. Intracellular bacteria were reduced >99% with 3+Pip-AcpP PPMO, whereas intracellular bacteria increased 3 orders of magnitude in untreated or Scr PPMO-treated cultures. We conclude that either AcpP PPMO or 3+Pip-AcpP PPMO inhibited growth of S. enterica in pure culture and that 3+Pip-AcpP PPMO reduced intracellular viability of S. enterica in macrophages.

* Corresponding author. Mailing address: Department of Microbiology, 220 Nash Hall, Oregon State University, Corvallis, OR 97331-3804. Phone: (541) 737-1845. Fax: (541) 737-0496. E-mail: gellerb{at}orst.edu

{triangledown} Published ahead of print on 6 July 2009.

{dagger} Present address: Department of Microbiology, 220 Nash Hall, Oregon State University, Corvallis, OR 97331-3804.

Antimicrobial Agents and Chemotherapy, September 2009, p. 3700-3704, Vol. 53, No. 9
0066-4804/09/$08.00+0     doi:10.1128/AAC.00099-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»