Abstract
Although much effort has been expended on evaluating recombinant proteins and synthetic peptides as immunogens, they have generally proved incapable of inducing an efficient cytotoxic T-cell (CTL) response. Filamentous bacteriophage fd can display multiple copies of foreign peptides in the N-terminal region of its major coat protein pVIII, 2,700 copies of which make up the virus capsid. Here we show that fd virions displaying peptide RT2 (ILKEPVHGV), corresponding to residues 309–317 of the reverse transcriptase (RTase) of HIV-1, are able to prime a CTL response specific for this HIV-1 epitope in human cell lines. Successful priming also requires a T-helper epitope, pep23 (KDSWTVNDIQKLVGK), corresponding to residues 249–263 of HIV-1 RTase. Supplying this by displaying it on either the same or a separate bacteriophage virion led to activation of antigen-specific CD4+ T cells. Likewise, HLA-A2 transgenic mice immunized with bacteriophage virions displaying peptide RT2 were shown to mount an effective, specific anti-HIV-RT2 CTL response. This unexpected ability to elicit a designated cytolytic T-cell response, in addition to a B-cell response, has important implications for access to the class I major histocompatibility complex (MHC) loading compartment and the development of recombinant vaccines.
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References
Stuhler, G. & Schlossman, S.F. Antigen organization regulates cluster formation and induction of cytotoxic T lymphocytes by helper T cell subsets. Proc. Natl. Acad. Sci. USA 94, 622–627 (1997).
Ossendorp, F., Mengede, E., Camps, M., Filius, R. & Melief, C. J. Specific T helper cell requirement for optimal induction of cytotoxic T lymphocytes against major histocompatibility complex class II negative tumors. J. Exp. Med. 187, 693–702 (1998).
Keene, J. A. & Forman, J. Helper activity is required for the in vivo generation of cytotoxic T lymphocytes. J. Exp. Med. 155, 768–782 (1982).
Bona, C.A., Casares, S. & Brumeanu, T.D. Towards development of T-cell vaccines. Immunol Today 19, 126–133 (1998).
Demotz, S., Grey, H. M. & Sette, A. The minimal number of class II MHC–antigen complexes needed for T cell activation. Science 249, 1028–1030 (1990).
Bot, A., Bot, S., Antohi, S., Karjalainen, K. & Bona, C. Kinetics of generation and persistence on membrane class II molecules of a viral peptide expressed on foreign and self proteins. J. Immunol. 157, 3436–3442 (1996).
Greenwood, J., Willis, A.E. & Perham, R.N. Multiple display of foreign peptides on a filamentous bacteriophage. Peptides from Plasmodium falciparum circumsporozoite protein as antigens. J. Mol. Biol. 220, 821–827 (1991).
Willis, A.E., Perham, R.N. & Wraith, D. Immunological properties of foreign peptides in multiple display on a filamentous bacteriophage. Gene 128, 79–83 (1993).
Minenkova, O.O., Ilyichev, A.A., Kishchenko, G.P. & Petrenko, V.A. Design of specific immunogens using filamentous phage as the carrier. Gene 128, 85–88 (1993).
di Marzo Veronese, F., Willis, A.E., Boyer-Thompson, C., Appella, E. & Perham, R.N. Structural mimicry and enhanced immunogenicity of peptide epitopes displayed on filamentous bacteriophage. The V3 loop of HIV-1 gp120. J. Mol. Biol. 243, 167–172 (1994).
De Berardinis, P. et al. Recognition of HIV-derived B and T cell epitopes displayed on filamentous phages. Vaccine 17, 1434–1441 (1999).
Ogg, G. S. et al. Quantitation of HIV-1-specific cytotoxic T lymphocytes and plasma load of viral RNA. Science 279, 2103–2106 (1998).
Malik, P. & Perham, R.N. New vectors for peptide display on the surface of filamentous bacteriophage. Gene 171, 49–51 (1996).
Manca, F. et al. Antigenicity of HIV-derived T helper determinants in the context of carrier recombinant proteins: effect on T helper cell repertoire selection. Eur. J. Immunol. 26, 2461–2469 (1996).
Menendez-Arias, L., Mas, A., Domingo, E. Cytotoxic T-lymphocyte responses to HIV-1 reverse transcriptase. Viral Immunol. 11, 167–181 (1998).
Malik, P. & Perham, R.N. Simultaneous display of different peptides on the surface of filamentous bacteriophage. Nucleic Acids Res. 25, 915–916 (1997).
Di Modugno, F. et al. MHC-peptide binding: dimers of cysteine-containing nonapeptides bind with high affinity to HLA-A2.1 class I molecules. J. Immunother 20, 431–436 (1997).
Ridge, J.P., Di Rosa, F. & Matzinger, P. A conditioned dendritic cell can be a temporal bridge between a CD4+ T- helper and a T-killer cell. Nature 393, 474–478 (1998).
Bennett, S.R. et al. Help for cytotoxic T-cell responses is mediated by CD40 signalling. Nature 393, 478–480 (1998).
Schoenberger, S.P., Toes, R.E., van der Voort, E.I., Offringa, R. & Melief, C.J. T-cell help for cytotoxic T lymphocytes is mediated by CD40–CD40L interactions. Nature 393, 480–483 (1998).
de Boer, M. et al. Ligation of B7 with CD28/CTLA-4 on T cells results in CD40 ligand expression, interleukin-4 secretion and efficient help for antibody production by B cells. Eur. J. Immunol 23, 3120–3125 (1993).
Pascolo, S. et al. HLA-A2.1-restricted education and cytolytic activity of CD8(+) T lymphocytes from beta2 microglobulin (beta2m) HLA-A2.1 monochain transgenic H-2Db beta2m double knockout mice. J. Exp. Med. 185, 2043–2051 (1997).
Vitiello, A., Marchesini, D., Furze, J., Sherman, L.A. & Chesnut, R.W. Analysis of the HLA-restricted influenza-specific cytotoxic T lymphocyte response in transgenic mice carrying a chimeric human–mouse class I major histocompatibility complex. J. Exp. Med. 173, 1007–1015 (1991).
Ren, J. et al. High resolution structures of HIV-1 RT from four RT–inhibitor complexes. Nat. Struct. Biol. 2, 293–302 (1995).
Grimison, B. & Laurence, J. Immunodominant epitope regions of HIV-1 reverse transcriptase: correlations with HIV-1+ serum IgG inhibitory to polymerase activity and with disease progression. J. Acquir. Immune. Defic. Syndr. Hum. Retrovirol. 9, 58–68 (1995).
Acknowledgements
This research was supported by a grant from the European Union to J.G. and R.N.P., from ISS II Progetto AIDS to J.G., and from CNR P.F. Biotecnologia to G.D.P. The core facilities of the Cambridge Centre for Molecular Recognition are supported by the Biotechnology and Biological Sciences Research Council and The Wellcome Trust. We thank P. Barba and C. Sole for excellent technical assistance.
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De Berardinis, P., Sartorius, R., Fanutti, C. et al. Phage display of peptide epitopes from HIV-1 elicits strong cytolytic responses. Nat Biotechnol 18, 873–876 (2000). https://doi.org/10.1038/78490
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DOI: https://doi.org/10.1038/78490
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