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A Novel Approach to Deliver a Mycobacterium avium subsp. paratuberculosis Antigen in Eukaryotic Cells

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Abstract

This study was aimed to express and deliver a Mycobacterium avium subsp. paratuberculosis antigen to macrophages using salmonella as carrier. The coding sequence of a fibronectin attachment protein which is expressed by Mycobacterium avium subsp. paratuberculosis was cloned into pcDNA3.1 (+) plasmid. The construct was introduced into the attenuated Salmonella typhimurium strain SL7207 (ΔhisG, ΔaroA) as carrier. In order to evaluate the delivery capacity of Salmonella and gene expression by antigen-presenting cells, the THP-1 derived macrophages were infected with the salmonella carrier. SDS-PAGE and western blot analysis showed the successful delivery and expression of targeted gene in THP-1 cell line. Although, in vitro stimulation of peripheral blood mononuclear cells with Salmonella containing plasmid did not trigger IFNγ production significantly. But it seems that this carrier can increase plasmid uptake and antigen expression by host intestinal antigen-presenting cells after mucosal administration. So, the construct can be used for further in vivo studies on the Salmonella carrier’s efficiency in mycobacterial DNA vaccines.

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Acknowledgements

The authors would like to thank Dr. Arsalan Hosseini for his valuable advice on the immunogenicity assay. This work was financially supported partially by a grant from Shiraz University and also Iran National Science Foundation (No: 94811303).

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Authors and Affiliations

  1. Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, 71345-1731, Iran

    Azar Motamedi Boroojeni, Abdollah Derakhshandeh, Masoud Haghkhah & Vida Eraghi

  2. Institute of Biotechnology and Bioengineering, Isfahan University of Technology, Esfahān, Iran

    Seyed Amir Hossein Jalali

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  1. Azar Motamedi Boroojeni
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  2. Abdollah Derakhshandeh
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  3. Masoud Haghkhah
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  4. Seyed Amir Hossein Jalali
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Correspondence to Abdollah Derakhshandeh.

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Ethical Approval

All animal procedures were performed in accordance with the guidelines of the Animal Ethics Committee of School of Veterinary Medicine, Shiraz University (dated 20 September 2013) and the experiments were approved by this committee (permit: 93GCU1M163973).

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Motamedi Boroojeni, A., Derakhshandeh, A., Haghkhah, M. et al. A Novel Approach to Deliver a Mycobacterium avium subsp. paratuberculosis Antigen in Eukaryotic Cells. Mol Biotechnol 61, 506–512 (2019). https://doi.org/10.1007/s12033-019-00180-8

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  • DOI: https://doi.org/10.1007/s12033-019-00180-8

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