Abstract
Antigen presentation by classical MHC class I molecules to CD8+ T cells is a central aspect of the adaptive immune response. Here, we describe methods to monitor antigen presentation using the model ovalbumin Kb-binding peptide, SIINFEKL. SIINFEKL genetically incorporated into viral or cellular source proteins can be used to precisely probe various aspects of antigen presentation, including the kinetics of peptide generation, MHC class I surface stability, and presentation efficiency following pharmacological and genetic manipulations including genome wide and high throughput drug screening.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Esquivel F, Yewdell JW, Bennink JR (1992) RMA/S cells present endogenously synthesized cytosolic proteins to class I-restricted cytotoxic T lymphocytes. J Exp Med 175:163–168
Yewdell JW, Anton LC, Bennink JR (1996) Defective ribosomal products (DRiPs): a major source of antigenic peptides for MHC class I molecules? J Immunol 157(5):1823–1826
Challa-Malladi M, Lieu YK, Califano O, Holmes AB, Bhagat G, Murty VV, Dominguez-Sola D, Pasqualucci L, Dalla-Favera R (2011) Combined genetic inactivation of beta2-Microglobulin and CD58 reveals frequent escape from immune recognition in diffuse large B cell lymphoma. Cancer Cell 20(6):728–740. https://doi.org/10.1016/j.ccr.2011.11.006
McGranahan N, Rosenthal R, Hiley CT, Rowan AJ, Watkins TBK, Wilson GA, Birkbak NJ, Veeriah S, Van Loo P, Herrero J, Swanton C (2017) Allele-specific HLA loss and immune escape in lung cancer evolution. Cell 171(6):1259–1271.e1211. https://doi.org/10.1016/j.cell.2017.10.001
Bassani-Sternberg M, Braunlein E, Klar R, Engleitner T, Sinitcyn P, Audehm S, Straub M, Weber J, Slotta-Huspenina J, Specht K, Martignoni ME, Werner A, Hein R, HB D, Peschel C, Rad R, Cox J, Mann M, Krackhardt AM (2016) Direct identification of clinically relevant neoepitopes presented on native human melanoma tissue by mass spectrometry. Nat Commun 7:13404. https://doi.org/10.1038/ncomms13404
Sendoel A, Dunn JG, Rodriguez EH, Naik S, Gomez NC, Hurwitz B, Levorse J, Dill BD, Schramek D, Molina H, Weissman JS, Fuchs E (2017) Translation from unconventional 5′ start sites drives tumour initiation. Nature 541(7638):494–499. https://doi.org/10.1038/nature21036
Yewdell JW, Bennink JR, Hosaka Y (1988) Cells process exogenous proteins for recognition by cytotoxic T lymphocytes. Science 239(4840):637–640
Moore MW, Carbone FR, Bevan MJ (1988) Introduction of soluble protein into the class I pathway of antigen processing and presentation. Cell 54:777–785
Karttunen J, Sanderson S, Shastri N (1992) Detection of rare antigen-presenting cells by the lacZ T-cell activation assay suggests an expression cloning strategy for T-cell antigens. Proc Natl Acad Sci U S A 89:6020–6024
Clarke SR, Barnden M, Kurts C, Carbone FR, Miller JF, Heath WR (2000) Characterization of the ovalbumin-specific TCR transgenic line OT-I: MHC elements for positive and negative selection. Immunol Cell Biol 78(2):110–117. https://doi.org/10.1046/j.1440-1711.2000.00889.x
Porgador A, Yewdell JW, Deng Y, Bennink JR, Germain RN (1997) Localization, quantitation, and in situ detection of specific peptide-MHC class I complexes using a monoclonal antibody. Immunity 6(6):715–726
Chen W, Anton LC, Bennink JR, Yewdell JW (2000) Dissecting the multifactorial causes of immunodominance in class I-restricted T cell responses to viruses. Immunity 12(1):83–93
Princiotta MF, Finzi D, Qian SB, Gibbs J, Schuchmann S, Buttgereit F, Bennink JR, Yewdell JW (2003) Quantitating protein synthesis, degradation, and endogenous antigen processing. Immunity 18(3):343–354
Mates L, Chuah MK, Belay E, Jerchow B, Manoj N, Acosta-Sanchez A, Grzela DP, Schmitt A, Becker K, Matrai J, Ma L, Samara-Kuko E, Gysemans C, Pryputniewicz D, Miskey C, Fletcher B, VandenDriessche T, Ivics Z, Izsvak Z (2009) Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. Nat Genet 41(6):753–761. https://doi.org/10.1038/ng.343
Kowarz E, Loscher D, Marschalek R (2015) Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol J 10(4):647–653. https://doi.org/10.1002/biot.201400821
Yewdell JW, Bennink JR (1989) Brefeldin A specifically inhibits presentation of protein antigens to cytotoxic T lymphocytes. Science 244(4908):1072–1075
del Val M, Schlicht HJ, Ruppert T, Reddehase MJ, Koszinowski UH (1991) Efficient processing of an antigenic sequence for presentation by MHC class I molecules depends on its neighboring residues in the protein. Cell 66:1145–1153
Zanker D, Waithman J, Yewdell JW, Chen W (2013) Mixed proteasomes function to increase viral peptide diversity and broaden antiviral CD8+ T cell responses. J Immunol 191(1):52–59. https://doi.org/10.4049/jimmunol.1300802
Acknowledgments
The authors are supported by the Division of Intramural Research, NIAID.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Dersh, D., Yewdell, J.W., Wei, J. (2019). A SIINFEKL-Based System to Measure MHC Class I Antigen Presentation Efficiency and Kinetics. In: van Endert, P. (eds) Antigen Processing. Methods in Molecular Biology, vol 1988. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9450-2_9
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9450-2_9
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9449-6
Online ISBN: 978-1-4939-9450-2
eBook Packages: Springer Protocols