Autotransporter-Based Antigen Display in Bacterial Ghosts

Applied and Environmental Microbiology - Tập 81 Số 2 - Trang 726-735 - 2015
Anna Hjelm1, Bill Söderström1, David Vikström2, Wouter S. P. Jong3,4, Joen Luirink3,4, Jan‐Willem de Gier1
1Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden
2Xbrane Bioscience AB, Stockholm, Sweden
3Abera Bioscience AB, Stockholm, Sweden
4Section Molecular Microbiology, Department of Molecular Cell Biology, Faculty of Earth and Life Sciences, VU University, Amsterdam, Netherlands

Tóm tắt

ABSTRACT Bacterial ghosts are empty cell envelopes of Gram-negative bacteria that can be used as vehicles for antigen delivery. Ghosts are generated by releasing the bacterial cytoplasmic contents through a channel in the cell envelope that is created by the controlled production of the bacteriophage ϕX174 lysis protein E. While ghosts possess all the immunostimulatory surface properties of the original host strain, they do not pose any of the infectious threats associated with live vaccines. Recently, we have engineered the Escherichia coli autotransporter hemoglobin protease (Hbp) into a platform for the efficient surface display of heterologous proteins in Gram-negative bacteria, HbpD. Using the Mycobacterium tuberculosis vaccine target ESAT6 (early secreted antigenic target of 6 kDa), we have explored the application of HbpD to decorate E. coli and Salmonella ghosts with antigens. The use of different promoter systems enabled the concerted production of HbpD-ESAT6 and lysis protein E. Ghost formation was monitored by determining lysis efficiency based on CFU, the localization of a set of cellular markers, fluorescence microscopy, flow cytometry, and electron microscopy. Hbp-mediated surface display of ESAT6 was monitored using a combination of a protease accessibility assay, fluorescence microscopy, flow cytometry and (immuno-)electron microscopy. Here, we show that the concerted production of HbpD and lysis protein E in E. coli and Salmonella can be used to produce ghosts that efficiently display antigens on their surface. This system holds promise for the development of safe and cost-effective vaccines with optimal intrinsic adjuvant activity and exposure of heterologous antigens to the immune system.

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Tài liệu tham khảo

10.4161/bbug.1.5.12540

10.1016/j.vaccine.2010.06.087

10.1586/14760584.6.2.241

10.1128/mr.56.3.430-481.1992

10.1016/S0021-9258(18)83778-4

10.1111/j.1365-2958.2012.08153.x

10.1021/bi900469g

10.1073/pnas.97.8.4297

10.1128/jb.172.7.4109-4114.1990

10.1046/j.1365-2958.1997.5781941.x

10.1111/j.1574-6976.1995.tb00203.x

10.1007/s12033-010-9277-2

10.1016/j.vaccine.2004.11.078

10.1073/pnas.93.4.1458

10.1016/S0928-8244(03)00063-4

10.1128/IAI.00950-10

10.1016/j.bbamcr.2013.11.006

10.1016/j.copbio.2010.07.009

10.1186/1475-2859-11-85

10.1128/iai.63.5.1710-1717.1995

10.1111/j.1365-2958.2007.05605.x

10.1186/1475-2859-5-23

10.1016/0022-2836(76)90119-4

10.1128/JB.181.2.508-520.1999

10.1038/291238a0

10.2144/000112112

10.1016/0378-1119(94)90643-2

10.1186/1475-2859-9-65

10.1073/pnas.120163297

10.1016/S0014-5793(01)02494-2

Lubitz W. June 2011. US patent 7 968 323 B2. http://www.google.com/patents/US7968323.pdf.

10.1074/mcp.M600431-MCP200

10.1002/1097-0320(20010601)44:2<106::AID-CYTO1088>3.0.CO;2-5

10.1128/AEM.69.1.468-474.2003

10.1046/j.1365-2958.1999.01425.x

10.1128/IAI.73.8.4810-4817.2005

10.1006/biol.1995.0034

10.1016/0378-1119(90)90464-3

10.1038/35101614

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Applied and Environmental Microbiology

Tập 81 Số 2

726-735

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