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Attenuated Salmonella enteritidis E23 as a vehicle for the rectal delivery of DNA vaccine coding for HIV-1 polyepitope CTL immunogen.

Karpenko LI, Danilenko AV, Bazhan SI, Danilenko ED, Sysoeva GM, Kaplina ON, Volkova OY, Oreshkova SF, Ilyichev AA - Microb Biotechnol (2011)

Bottom Line: The gene coding for TCI protein was used to construct the eukaryotic expression plasmid pcDNA-TCI.Detectable antibodies were generated in 2 weeks after immunization and their level increased after second immunization.This study demonstrates that attenuated S. enteritidis E23 is an effective live vector for rectal delivery of the DNA vaccine pcDNA-TCI to generate humoral and T-cellular responses against HIV-1.

View Article: PubMed Central - PubMed

Affiliation: State Research Center of Virology and Biotechnology 'Vector', 630559 Koltsovo, Novosibirsk, Russia. lkarpenko@ngs.ru

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RT‐PCR detection of in vivo TCI gene transcription. Electrophoretic pattern of PCR products in 1% agarose gel: lane M, λ/StyI; length of fragments are 19 329, 7743, 6223, 4254, 3472, 2690, 1882, 1489, 925, 421 and 74 bp; lane 1, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA‐TCI as a template; and lane 2, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA3.1 as a template.
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f3: RT‐PCR detection of in vivo TCI gene transcription. Electrophoretic pattern of PCR products in 1% agarose gel: lane M, λ/StyI; length of fragments are 19 329, 7743, 6223, 4254, 3472, 2690, 1882, 1489, 925, 421 and 74 bp; lane 1, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA‐TCI as a template; and lane 2, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA3.1 as a template.

Mentions: Peyer's patches are the main colonization site of the attenuated S. enteritidis E23/pcDNA‐TCI and an important immunologically relevant site in the context of the mucosal responsiveness. To test an in vivo delivery of pcDNA‐TCI using attenuated Salmonella as a transgenic vehicle, total cellular RNA was isolated from mouse small intestinal Peyer's patches on day 3 after the immunization with attenuated S. enteritidis E23/pcDNA‐TCI and the TCI gene transcription was analysed by RT‐PCR. As it is shown in (Fig. 3, only a DNA fragment with a length of about 1191 bp was amplified from RNA of the mice immunized with S. enteritidis E23/pcDNA‐TCI. Meanwhile, there were no DNA fragments amplified from the RNA prior to reverse transcription with the same primers or from the RNA of the control mice immunized with S. enteritidis E23/pcDNA3.1. Murine β‐actin DNA fragment (330 bp) was amplified from all samples.


Attenuated Salmonella enteritidis E23 as a vehicle for the rectal delivery of DNA vaccine coding for HIV-1 polyepitope CTL immunogen.

Karpenko LI, Danilenko AV, Bazhan SI, Danilenko ED, Sysoeva GM, Kaplina ON, Volkova OY, Oreshkova SF, Ilyichev AA - Microb Biotechnol (2011)

RT‐PCR detection of in vivo TCI gene transcription. Electrophoretic pattern of PCR products in 1% agarose gel: lane M, λ/StyI; length of fragments are 19 329, 7743, 6223, 4254, 3472, 2690, 1882, 1489, 925, 421 and 74 bp; lane 1, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA‐TCI as a template; and lane 2, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA3.1 as a template.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815784&req=5

f3: RT‐PCR detection of in vivo TCI gene transcription. Electrophoretic pattern of PCR products in 1% agarose gel: lane M, λ/StyI; length of fragments are 19 329, 7743, 6223, 4254, 3472, 2690, 1882, 1489, 925, 421 and 74 bp; lane 1, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA‐TCI as a template; and lane 2, the PCR product obtained using the RNA from the mice immunized with S. enteritidis E23/pcDNA3.1 as a template.
Mentions: Peyer's patches are the main colonization site of the attenuated S. enteritidis E23/pcDNA‐TCI and an important immunologically relevant site in the context of the mucosal responsiveness. To test an in vivo delivery of pcDNA‐TCI using attenuated Salmonella as a transgenic vehicle, total cellular RNA was isolated from mouse small intestinal Peyer's patches on day 3 after the immunization with attenuated S. enteritidis E23/pcDNA‐TCI and the TCI gene transcription was analysed by RT‐PCR. As it is shown in (Fig. 3, only a DNA fragment with a length of about 1191 bp was amplified from RNA of the mice immunized with S. enteritidis E23/pcDNA‐TCI. Meanwhile, there were no DNA fragments amplified from the RNA prior to reverse transcription with the same primers or from the RNA of the control mice immunized with S. enteritidis E23/pcDNA3.1. Murine β‐actin DNA fragment (330 bp) was amplified from all samples.

Bottom Line: The gene coding for TCI protein was used to construct the eukaryotic expression plasmid pcDNA-TCI.Detectable antibodies were generated in 2 weeks after immunization and their level increased after second immunization.This study demonstrates that attenuated S. enteritidis E23 is an effective live vector for rectal delivery of the DNA vaccine pcDNA-TCI to generate humoral and T-cellular responses against HIV-1.

View Article: PubMed Central - PubMed

Affiliation: State Research Center of Virology and Biotechnology 'Vector', 630559 Koltsovo, Novosibirsk, Russia. lkarpenko@ngs.ru

Show MeSH
Related in: MedlinePlus