Abstract
The identification and analysis of allergen-specific CD4+ T cells is critical for understanding how these cells contribute to atopic disease and how to subvert their behavior through immune therapy. The advent of fluorescently labeled soluble tetramers of peptide:MHCII complexes (pMHCII tetramers) has provided investigators with an invaluable means to achieve this goal. Although pMHCII tetramers were first developed over two decades ago, their widespread use has been limited by the technical difficulty of generating these reagents. However, the adoption of various technical innovations from several labs over time has contributed greatly to the increased success in tetramer generation today. Here, we describe a comprehensive protocol for generating pMHCII tetramers using as an example a Derp1:I-Ab tetramer used to study allergen-specific CD4+ T cell responses in murine models of airway inflammation and allergic disease.
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
Similar content being viewed by others
References
Locksley RM (2010) Asthma and allergic inflammation. Cell 140(6):777–783
Nepom GT (2012) MHC class II tetramers. J Immunol 188(6):2477–2482
Chua KY et al (1988) Sequence analysis of cDNA coding for a major house dust mite allergen, Der p 1. Homology with cysteine proteases. J Exp Med 167(1):175–182
Yssel H et al (1992) T cell activation-inducing epitopes of the house dust mite allergen Der p I. Proliferation and lymphokine production patterns by Der p I-specific CD4+ T cell clones. J Immunol 148(3):738–745
Hoyne GF, Callow MG, Kuo MC, Thomas WR (1994) Inhibition of T-cell responses by feeding peptides containing major and cryptic epitopes: studies with the Der p I allergen. Immunology 83(2):190–195
Hondowicz BD et al (2016) Interleukin-2-dependent allergen-specific tissue- resident memory cells drive asthma. Immunity 44(1):155–166
Jenkins MK, Moon JJ (2012) The role of naive T cell precursor frequency and recruitment in dictating immune response magnitude. J Immunol 188(9):4135–4140
Moon JJ et al (2007) Naive CD4(+) T cell frequency varies for different epitopes and predicts repertoire diversity and response magnitude. Immunity 27(2):203–213
Legoux FP, Moon JJ (2012) Peptide:MHC tetramer-based enrichment of epitope-specific T cells. J Vis Exp 68:e4420
Altman JD et al (1996) Phenotypic analysis of antigen-specific T lymphocytes. Science 274(5284):94–96
Vollers SS, Stern LJ (2008) Class II major histocompatibility complex tetramer staining: progress, problems, and prospects. Immunology 123(3):305–313
Scott CA, Garcia KC, Carbone FR, Wilson IA, Teyton L (1996) Role of chain pairing for the production of functional soluble IA major histocompatibility complex class II molecules. J Exp Med 183(5):2087–2095
Kozono H, White J, Clements J, Marrack P, Kappler J (1994) Production of soluble MHC class II proteins with covalently bound single peptides. Nature 369(6476):151–154
Beckett D, Kovaleva E, Schatz PJ (1999) A minimal peptide substrate in biotin holoenzyme synthetase-catalyzed biotinylation. Protein Sci 8(4):921–929
Yang J, Jaramillo A, Shi R, Kwok WW, Mohanakumar T (2004) In vivo biotinylation of the major histocompatibility complex (MHC) class II/peptide complex by coexpression of BirA enzyme for the generation of MHC class II/tetramers. Hum Immunol 65(7):692–699
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Moon, J.J., Pepper, M. (2018). Generation of Allergen-Specific Tetramers for a Murine Model of Airway Inflammation. In: Reinhardt, R. (eds) Type 2 Immunity. Methods in Molecular Biology, vol 1799. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7896-0_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7896-0_14
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-7895-3
Online ISBN: 978-1-4939-7896-0
eBook Packages: Springer Protocols