Improved delivery of the OVA-CD4 peptide to T helper cells by polymeric surface display on Salmonella

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• 作者：Junjie Zhang (10) (8) (9)
  Leon De Masi (9)
  Beena John (9)
  Wenxin Chen (8)
  Dieter M Schifferli (9)
  
  10. College of Food and Biological Engineering ; Zhengzhou University of Light Industry ; Zhengzhou ; Henan province ; 450002 ; People鈥檚 Republic of China
  8. State Key Laboratory of Agrobiotechnology and College of Biological Sciences ; China Agricultural University ; Beijing ; 100193 ; People鈥檚 Republic of China
  9. Department of Pathobiology ; University of Pennsylvania School of Veterinary Medicine ; Philadelphia ; Pennsylvania ; 19104 ; USA
  
• 关键词：Autotransporter ; T5SS ; Salmonella ; MisL ; Ovalbumin
• 刊名：Microbial Cell Factories
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：13
• 期：1
• 全文大小：916 KB
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• 刊物类别：Chemistry and Materials Science
• 刊物主题：Biotechnology
  Applied Microbiology
  Environmental Engineering/Biotechnology
  
• 出版者：BioMed Central
• ISSN：1475-2859

文摘

Background Autotransporter proteins represent a treasure trove for molecular engineers who modify Gram-negative bacteria for the export or secretion of foreign proteins across two membrane barriers. A particularly promising direction is the development of autotransporters as antigen display or secretion systems. Immunologists have been using ovalbumin as a reporter antigen for years and have developed sophisticated tools to detect specific T cells that respond to ovalbumin. Although ovalbumin-expressing bacteria are being used to trace T cell responses to colonizing or invading pathogens, current constructs for ovalbumin presentation have not been optimized. Results The activation of T helper cells in response to ovalbumin was improved by displaying the OVA-CD4 reporter epitope as a multimer on the surface of Salmonella and fused to the autotransporter MisL. Expression was optimized by including tandem in vivo promoters and two post-segregational killing systems for plasmid stabilization. Conclusions The use of an autotransporter protein to present relevant epitope repeats on the surface of bacteria, combined with additional techniques favoring stable and efficient in vivo transcription, optimizes antigen presentation to T cells. The technique of multimeric epitope surface display should also benefit the development of new Salmonella or other enterobacterial vaccines.