Processing of a Multiple Membrane Spanning Epstein-Barr Virus Protein for Cd8+T Cell Recognition Reveals a Proteasome-Dependent, Transporter Associated with Antigen Processing-Independent Pathway

doi:10.1084/jem.194.8.1053

Published 15 October 2001. doi:10.1084/jem.194.8.1053

This Article

	Full Text
	Full Text (PDF, 317K)
	PPT slides of all figures
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JEM
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Lautscham, G.
	Articles by Blake, N.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Lautscham, G.
	Articles by Blake, N.


Social Bookmarking

	What's this?

© The Rockefeller University Press, 0022-1007/2001/10/1053/ $5.00
The Journal of Experimental Medicine, Volume 194, Number 8, October 15, 2001 1053-1068

Original Article

Processing of a Multiple Membrane Spanning Epstein-Barr Virus Protein for Cd8⁺T Cell Recognition Reveals a Proteasome-Dependent, Transporter Associated with Antigen Processing–Independent Pathway

Georg Lautscham^a, Sabine Mayrhofer^a, Graham Taylor^a, Tracey Haigh^a, Alison Leese^a, Alan Rickinson^a, and Neil Blake^a

^a Cancer Research Campaign Institute for Cancer Studies and Medical Research Council Centre for Immune Regulation, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom
CRC Institute for Cancer Studies, University of Birmingham, Vincent Drive, Edgbaston, Birmingham B15 2TT, UK.44-121-414-448644-121-414-4492

A.B.Rickinson{at}bham.ac.uk

Epstein-Barr virus (EBV) latent membrane protein (LMP)2 is amultiple membrane spanning molecule which lacks ectodomainsprojecting into the lumen of the endoplasmic reticulum (ER).Human CD8⁺ cytotoxic T lymphocytes (CTL)s recognize a numberof epitopes within LMP2. Assays with epitope-specific CTLs intwo different cell backgrounds lacking the transporter associatedwith antigen processing (TAP) consistently show that some, butnot all, LMP2 epitopes are presented in a TAP-independent manner.However, unlike published examples of TAP-independent processingfrom endogenously expressed antigens, presentation of TAP-independentLMP2 epitopes was abrogated by inhibition of proteasomal activity.We found a clear correlation between hydrophobicity of the LMP2epitope sequence and TAP independence, and experiments withvaccinia minigene constructs expressing cytosolic epitope peptidesconfirmed that these more hydrophobic peptides were selectivelyable to access the HLA class I pathway in TAP-negative cells.Furthermore, the TAP-independent phenotype of particular epitopesequences did not require membrane location of the source antigensince (i) TAP-independent LMP2 epitopes inserted into an EBVnuclear antigen and (ii) hydrophobic epitope sequences nativeto EBV nuclear antigens were both presented in TAP-negativecells. We infer that there is a proteasome-dependent, TAP-independentpathway of antigen presentation which hydrophobic epitopes canselectively access.

Key Words: MHC class I presentation • CD8⁺ epitopes • hydrophobicity • Epstein-Barr virus • TAP independence

Abbreviations used in this paper: EBV, Epstein-Barr virus; ER,endoplasmic reticulum; LCL, lymphoblastoid cell line; LMP, latentmembrane protein; moi, multiplicity of infection; TAP, transporterassociated with antigen processing.

S. Mayrhofer's present address is Universitaet Koeln, Hodgkinlabor,LFI, D-50924 Koeln, Germany.

N. Blake's present address is Dept. of Medical Microbiologyand GU Medicine, University of Liverpool, Liverpool L69 3GA,United Kingdom.

Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Bell, M. J., Abbott, R. J. M., Croft, N. P., Hislop, A. D., Burrows, S. R. (2009). An HLA-A2-Restricted T-Cell Epitope Mapped to the BNLF2a Immune Evasion Protein of Epstein-Barr Virus That Inhibits TAP. J. Virol. 83: 2783-2788 [Abstract] [Full Text]
Johnstone, C., Guil, S., Rico, M. A., Garcia-Barreno, B., Lopez, D., Melero, J. A., Del Val, M. (2008). Relevance of viral context and diversity of antigen-processing routes for respiratory syncytial virus cytotoxic T-lymphocyte epitopes. J. Gen. Virol. 89: 2194-2203 [Abstract] [Full Text]
Huang, S., Gilfillan, S., Kim, S., Thompson, B., Wang, X., Sant, A. J., Fremont, D. H., Lantz, O., Hansen, T. H. (2008). MR1 uses an endocytic pathway to activate mucosal-associated invariant T cells. JEM 205: 1201-1211 [Abstract] [Full Text]
Zuo, J., Thomas, W., van Leeuwen, D., Middeldorp, J. M., Wiertz, E. J. H. J., Ressing, M. E., Rowe, M. (2008). The DNase of Gammaherpesviruses Impairs Recognition by Virus-Specific CD8+ T Cells through an Additional Host Shutoff Function. J. Virol. 82: 2385-2393 [Abstract] [Full Text]
Haigh, T. A., Lin, X., Jia, H., Hui, E. P., Chan, A. T. C., Rickinson, A. B., Taylor, G. S. (2008). EBV Latent Membrane Proteins (LMPs) 1 and 2 as Immunotherapeutic Targets: LMP-Specific CD4+ Cytotoxic T Cell Recognition of EBV-Transformed B Cell Lines. J. Immunol. 180: 1643-1654 [Abstract] [Full Text]
Oosten, L. E. M., Koppers-Lalic, D., Blokland, E., Mulder, A., Ressing, M. E., Mutis, T., van Halteren, A. G. S., Wiertz, E. J. H. J., Goulmy, E. (2007). TAP-inhibiting proteins US6, ICP47 and UL49.5 differentially affect minor and major histocompatibility antigen-specific recognition by cytotoxic T lymphocytes. Int Immunol 19: 1115-1122 [Abstract] [Full Text]
Herberts, C. A., Neijssen, J. J., Haan, J. d., Janssen, L., Drijfhout, J. W., Reits, E. A., Neefjes, J. J. (2006). Cutting Edge: HLA-B27 Acquires Many N-Terminal Dibasic Peptides: Coupling Cytosolic Peptide Stability to Antigen Presentation.. J. Immunol. 176: 2697-2701 [Abstract] [Full Text]
Ito, Y., Kondo, E., Demachi-Okamura, A., Akatsuka, Y., Tsujimura, K., Tanimoto, M., Morishima, Y., Takahashi, T., Kuzushima, K. (2006). Three Immunoproteasome-Associated Subunits Cooperatively Generate a Cytotoxic T-Lymphocyte Epitope of Epstein-Barr Virus LMP2A by Overcoming Specific Structures Resistant to Epitope Liberation. J. Virol. 80: 883-890 [Abstract] [Full Text]
Dorfel, D., Appel, S., Grunebach, F., Weck, M. M., Muller, M. R., Heine, A., Brossart, P. (2005). Processing and presentation of HLA class I and II epitopes by dendritic cells after transfection with in vitro-transcribed MUC1 RNA. Blood 105: 3199-3205 [Abstract] [Full Text]
Hill, S. C., Youde, S. J., Man, S., Teale, G. R., Baxendale, A. J., Hislop, A., Davies, C. C., Luesley, D. M., Blom, A. M., Rickinson, A. B., Young, L. S., Eliopoulos, A. G. (2005). Activation of CD40 in Cervical Carcinoma Cells Facilitates CTL Responses and Augments Chemotherapy-Induced Apoptosis. J. Immunol. 174: 41-50 [Abstract] [Full Text]
Lee, S. P., Brooks, J. M., Al-Jarrah, H., Thomas, W. A., Haigh, T. A., Taylor, G. S., Humme, S., Schepers, A., Hammerschmidt, W., Yates, J. L., Rickinson, A. B., Blake, N. W. (2004). CD8 T Cell Recognition of Endogenously Expressed Epstein-Barr Virus Nuclear Antigen 1. JEM 199: 1409-1420 [Abstract] [Full Text]
Taylor, G. S., Haigh, T. A., Gudgeon, N. H., Phelps, R. J., Lee, S. P., Steven, N. M., Rickinson, A. B. (2004). Dual Stimulation of Epstein-Barr Virus (EBV)-Specific CD4+- and CD8+-T-Cell Responses by a Chimeric Antigen Construct: Potential Therapeutic Vaccine for EBV-Positive Nasopharyngeal Carcinoma. J. Virol. 78: 768-778 [Abstract] [Full Text]
Edwards, R. H., Sitki-Green, D., Moore, D. T., Raab-Traub, N. (2004). Potential Selection of LMP1 Variants in Nasopharyngeal Carcinoma. J. Virol. 78: 868-881 [Abstract] [Full Text]
Majlessi, L., Rojas, M.-J., Brodin, P., Leclerc, C. (2003). CD8+-T-Cell Responses of Mycobacterium-Infected Mice to a Newly Identified Major Histocompatibility Complex Class I-Restricted Epitope Shared by Proteins of the ESAT-6 Family. Infect. Immun. 71: 7173-7177 [Abstract] [Full Text]
Ong, K.-W., Wilson, A. D., Hirst, T. R., Morgan, A. J. (2003). The B Subunit of Escherichia coli Heat-Labile Enterotoxin Enhances CD8+ Cytotoxic-T-Lymphocyte Killing of Epstein-Barr Virus-Infected Cell Lines. J. Virol. 77: 4298-4305 [Abstract] [Full Text]
Lautscham, G., Haigh, T., Mayrhofer, S., Taylor, G., Croom-Carter, D., Leese, A., Gadola, S., Cerundolo, V., Rickinson, A., Blake, N. (2003). Identification of a TAP-Independent, Immunoproteasome-Dependent CD8+ T-Cell Epitope in Epstein-Barr Virus Latent Membrane Protein 2. J. Virol. 77: 2757-2761 [Abstract] [Full Text]
Kuzushima, K., Hayashi, N., Kudoh, A., Akatsuka, Y., Tsujimura, K., Morishima, Y., Tsurumi, T. (2003). Tetramer-assisted identification and characterization of epitopes recognized by HLA A*2402-restricted Epstein-Barr virus-specific CD8+ T cells. Blood 101: 1460-1468 [Abstract] [Full Text]
Heinzel, A. S., Grotzke, J. E., Lines, R. A., Lewinsohn, D. A., McNabb, A. L., Streblow, D. N., Braud, V. M., Grieser, H. J., Belisle, J. T., Lewinsohn, D. M. (2002). HLA-E-dependent Presentation of Mtb-derived Antigen to Human CD8+ T Cells. JEM 196: 1473-1481 [Abstract] [Full Text]