Zeng, X. W., Zhang, Kefeng, Li, C. Y., Zhou, Yu, Bo, L., Zhang, X. M. and Hong, Yiguo . (2008) Expression of highly immunogenic tuberculosis proteins in plants using a versatile Potato virus X-based expression system. Journal of Horticultural Science and Biotechnology, Vol.83 (No.1). pp. 4-8. ISSN 1462-0316

Full text not available from this repository.

Abstract

The functionally and structurally-coordinated proteins ESAT-6 and CFP-10 secreted by Mycobacterium tuberculosis are potent T-cell antigens and hold great promise as novel subunit vaccines against tuberculosis (TB). Using a modified Potato virus X, we expressed the ESAT-6 and CFP-10 proteins in Nicotiana benthamiana and in tomato plants. Both proteins were tagged with a HiS(6)-Gly polypeptide at their N-termini, while a green fluorescent protein (GFP) was fused in-frame at their C-termini. Our data suggest that free ESAT-6 and CFP-10 were less stable than the GFP fusion proteins in plant cells. GFP-tagging significantly increased the stability of both proteins, as shown by the fact that ESAT6:GFP and CFP10:GFP were readily detected by western blotting. The GFP tag also allowed direct visualisation and localisation of green fluorescence, an efficient reporter of fusion protein expression in those plant tissues infected by the recombinant viruses. The potential for producing ESAT-6 and CFP-10 as novel orally-deliverable TB vaccines in tomato fruit is discussed.

Item Type:	Journal Article
Subjects:	Q Science > QK Botany Q Science > QR Microbiology S Agriculture > SB Plant culture
Divisions:	Faculty of Science > Life Sciences (2010- ) > Warwick HRI (2004-2010)
Library of Congress Subject Headings (LCSH):	Tuberculosis -- Molecular aspects, Immune response, Proteins, Potato virus X , Mycobacterium tuberculosis
Journal or Publication Title:	Journal of Horticultural Science and Biotechnology
Publisher:	Headley Bros. Ltd.
ISSN:	1462-0316
Date:	January 2008
Volume:	Vol.83
Number:	No.1
Number of Pages:	5
Page Range:	pp. 4-8
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	Chengdu Institute of Biological Products, Warwick HRI
References:	BERTHET, F. X., RASMUSSEN, P. B., ROSENKRANDS, I., ANDERSEN, P. and GICQUEL, B. (1998). A Mycobacterium tuberculosis operon encoding ESAT-6 and a novel low-molecular-mass culture filtrate protein (CFP-10).Microbiology, 144, 3195–3203. BRANDT, L., ELHAY, M., ROSENKRANDS, I., LINDBLAD, E.B. and ANDERSEN, P. (2000). ESAT-6 subunit vaccination against Mycobacterium tuberculosis. Infection and Immunity, 68, 791–795. BRODIN, P., MAJLESSI, L., MARSOLLIER, L., DE JONGE, M. I., BOTTAI, D., DEMANGEL, C., HINDS, J., NEYROLLES, O., BUTCHER, P.D., LECLERC,C.,COLE, S.T. and BROSCH, R. (2006). Dissection of ESAT-6 system 1 of Mycobacterium tuberculosis and impact on immunogenicity and virulence. Infection and Immunity, 74, 88–98. DOROKHOV,Y.L.,SHEVELEVA,A.A.,FROLOVA,O.Y.,KOMAROVA,T.V., ZVEREVA, A. S., IVANOV, P. A. and ATABEKOV, J. G. (2007). Superexpression of tuberculosis antigens in plant leaves. Tuberculosis, 87, 218–224. FINE, P.E.M. (1995).Variation in protection by BCG: implication of and for heterologous immunity. Lancet, 346, 1339–1345. FOX, J. L. (2006). Turning plants into protein factories. Nature Biotechnology, 24, 1191–1193. GUPTA,U.D.,KATOCH,V.M. and MCMURRAY,D.N. (2007).Current status of TB vaccines. Vaccine, 25, 3742–3751. HONG, Y., SAUNDERS, K., HARTLEY, M. R. and STANLEY, J. (1996). Resistance to geminivirus infection by virus-induced expression of dianthin in transgenic plants. Virology, 220, 119–127. KAMATH, A. T., FENG, C. G., MACDONALD, M., BRISCOE, H. and BRITTON,W. J. (1999). Differential protective efficacy of DNA vaccines expressing secreted proteins of Mycobacterium tuberculosis. Infection and Immunity, 67, 1702–1707. LI, Z., HOWARD, A., KELLEY, C., DELOGU, G., COLLINS, F. and MORRIS, S. (1999). Immunogenicity of DNA vaccines expressing tuberculosis proteins fused to tissue plasminogen activator signal sequences. Infection and Immunity, 67, 4780–4786. MA, J.K-C.,CHIKWAMBA,R., SPARROW, P., FISCHER,R.,MAHONEY,R. and TWYMAN, R. M. (2005). Plant-derived pharmaceuticals – the road forward. Trends in Plant Sciences, 10, 580–585. MANNING, K., TOR, M., POOLE, M., HONG, Y., THOMPSON, A. J., KING, G. J., GIOVANNONI, J. J. and SEYMOUR, G. B. (2006). A naturally occurring epigenetic mutation in a gene encoding an SBP-box transcription factor inhibits tomato fruit ripening. Nature Genetics, 38, 948–952. PORTA, C. and LOMONOSSOFF,G. P. (2002).Viruses as vectors for the expression of foreign sequences in plants. Biotechnology and Genetic Engineering Review, 19, 245–291. PYM, A. S., BRODIN, P., MAJLESSI, L., BROSCH, R., DEMANGEL, C., WILLIAMS, A.,GRIFFITHS, K. E.,MARCHAL,G., LECLERC, C. and COLE, S. T. (2003). Recombinant BCG exporting ESAT-6 confers enhanced protection against tuberculosis. Nature Medicine, 9, 533–539. RENSHAW, P. S., PANAGIOTIDOU, P., WHELAN, A., GORDON, S. V., HEWINSON, R. G.,WILLIAMSON, R. A. and CARR, M. D. (2002). Conclusive evidence that the major T-cell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight, 1:1 complex and characterization of the structural properties of ESAT-6, CFP-10, and the ESAT-6/CFP-10 complex. Implications for pathogenesis and virulence. Journal of Biological Chemistry, 277, 21598–21603. RENSHAW, P. S., LIGHTBODY, K. L., VEVERKA, V., MUSKETT, F. W., KELLY, G., FRENKIEL, T. A., GORDON, S. V., HEWINSON, R. G., BURKE, B., NORMAN, J., WILLIAMSON, R. A. and CARR, M. D. (2005). Structure and function of the complex formed by the tuberculosis virulence factors CFP-10 and ESAT-6. EMBO Journal, 24, 2491–2498. RIGANO, M. M., ALVAREZ, M. L., PINKHASOV, J., JIN, Y., SALA, F., ARNTZEN, C. J. and WALMSLEY, A. M. (2004). Production of a fusion protein consisting of the enterotoxigenic Escherichia coli heat-labile toxin B subunit and a tuberculosis antigen in Arabidopsis thaliana. Plant Cell Reports, 22, 502–508. RIGANO, M. M., DREITZ, S., KIPNIS, A. P., IZZO, A. A. and WALMSLEY, A. M. (2006). Oral immunogenicity of a plant-made, subunit, tuberculosis vaccine. Vaccine, 24, 691–695. RYABOV, E. V., VAN WEZEL, R., WALSH, J. and HONG, Y. (2004). Cell-to-cell, but not long-distance, spread of RNA silencing that is induced in individual epidermal cells. Journal of Virology, 78, 3149–3154. SKEIKY,Y.A.W. and SADOFF, J. C. (2006).Advances in tuberculosis vaccine strategies. Nature Reviews Microbiology, 4, 469–476. SKJOT, R. L., OETTINGER, T., ROSENKRANDS, I., RAVN, P., BROCK, I., JACOBSEN, S. and ANDERSEN, P. (2000). Comparative evaluation of low-molecular-mass proteins from Mycobacterium tuberculosis identifies members of the ESAT-6 family as immunodominant T-cell antigens. Infection and Immunity, 68, 214–220. VAN WEZEL, R. and HONG, Y. (2004). Virus survival of RNA silencing without deploying protein-mediated suppression in Nicotiana benthamiana. FEBS Letters, 562, 65–70. VAN WEZEL, R., LIU, H., TIEN, P., STANLEY, J. and HONG, Y. (2001) Gene C2 of the monopartite geminivirus tomato yellow leaf curl virus-China encodes a pathogenicity determinant that is localized in the nucleus. Molecular Plant-Microbe Interactions, 14, 1125–1128. WARDS, B. J., DE LISLE, G W. and COLLINS, D. M. (2000). An esat6 knockout mutant of Mycobacterium bovis produced by homologous recombination will contribute to the development of a live tuberculosis vaccine. Tuberculosis Lung Diseases, 80, 185–189. ZELADA, A. M., CALAMANTE, G., DE LA PAZ SANTANGELO, M., BIGI, F., VERNA, F., MENTABERRY, A. and CATALDI, A. (2006). Expression of tuberculosis antigen ESAT-6 in Nicotiana tabacum using a potato virus X-based vector. Tuberculosis, 86, 263–267.
URI:	http://wrap.warwick.ac.uk/id/eprint/30621

Data sourced from Thomson Reuters' Web of Knowledge

Request changes to a record

Actions (login required)

View Item