Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions

Peter Jönsson; Jennifer H Southcombe; Ana Mafalda Santos; Jiandong Huo; Ricardo A Fernandes; James McColl; Melissa Lever; Edward J Evans; Alexander Hudson; Veronica T Chang; Tomáš Hanke; Andrew Godkin; Paul D Dunne; Mathew H Horrocks; Matthieu Palayret; Gavin R Screaton; Jan Petersen; Jamie Rossjohn; Lars Fugger; Omer Dushek; Xiao-Ning Xu; Simon J Davis; David Klenerman

doi:10.1073/pnas.1513918113

Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions

Peter Jönsson, Jennifer H Southcombe, Ana Mafalda Santos, Jiandong Huo, Ricardo A Fernandes, James McColl, Melissa Lever, Edward J Evans, Alexander Hudson, Veronica T Chang, Tomáš Hanke, Andrew Godkin, Paul D Dunne, Mathew H Horrocks, Matthieu Palayret, Gavin R Screaton, Jan Petersen, Jamie Rossjohn, Lars Fugger, Omer DushekXiao-Ning Xu, Simon J Davis, David Klenerman

Institut for Klinisk Medicin - Immunologi

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

Abstract

The αβ T-cell coreceptor CD4 enhances immune responses more than 1 million-fold in some assays, and yet the affinity of CD4 for its ligand, peptide-major histocompatibility class II (pMHC II) on antigen-presenting cells, is so weak that it was previously unquantifiable. Here, we report that a soluble form of CD4 failed to bind detectably to pMHC II in surface plasmon resonance-based assays, establishing a new upper limit for the solution affinity at 2.5 mM. However, when presented multivalently on magnetic beads, soluble CD4 bound pMHC II-expressing B cells, confirming that it is active and allowing mapping of the native coreceptor binding site on pMHC II. Whereas binding was undetectable in solution, the affinity of the CD4/pMHC II interaction could be measured in 2D using CD4- and adhesion molecule-functionalized, supported lipid bilayers, yielding a 2D Kd of ∼5,000 molecules/μm(2) This value is two to three orders of magnitude higher than previously measured 2D Kd values for interacting leukocyte surface proteins. Calculations indicated, however, that CD4/pMHC II binding would increase rates of T-cell receptor (TCR) complex phosphorylation by threefold via the recruitment of Lck, with only a small, 2-20% increase in the effective affinity of the TCR for pMHC II. The affinity of CD4/pMHC II therefore seems to be set at a value that increases T-cell sensitivity by enhancing phosphorylation, without compromising ligand discrimination.

Originalsprog	Engelsk
Tidsskrift	Proceedings of the National Academy of Sciences
Vol/bind	113
Nummer	20
Sider (fra-til)	5682-7
Antal sider	6
ISSN	0027-8424
DOI	https://doi.org/10.1073/pnas.1513918113
Status	Udgivet - 17 maj 2016

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10.1073/pnas.1513918113

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4878507/pdf/pnas.201513918.pdf

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Jönsson, P., Southcombe, J. H., Santos, A. M., Huo, J., Fernandes, R. A., McColl, J., Lever, M., Evans, E. J., Hudson, A., Chang, V. T., Hanke, T., Godkin, A., Dunne, P. D., Horrocks, M. H., Palayret, M., Screaton, G. R., Petersen, J., Rossjohn, J., Fugger, L., ... Klenerman, D. (2016). Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions. Proceedings of the National Academy of Sciences, 113(20), 5682-7. https://doi.org/10.1073/pnas.1513918113

@article{f073b676c1b54ec98134ec94e1b1f6a9,

title = "Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions",

abstract = "The αβ T-cell coreceptor CD4 enhances immune responses more than 1 million-fold in some assays, and yet the affinity of CD4 for its ligand, peptide-major histocompatibility class II (pMHC II) on antigen-presenting cells, is so weak that it was previously unquantifiable. Here, we report that a soluble form of CD4 failed to bind detectably to pMHC II in surface plasmon resonance-based assays, establishing a new upper limit for the solution affinity at 2.5 mM. However, when presented multivalently on magnetic beads, soluble CD4 bound pMHC II-expressing B cells, confirming that it is active and allowing mapping of the native coreceptor binding site on pMHC II. Whereas binding was undetectable in solution, the affinity of the CD4/pMHC II interaction could be measured in 2D using CD4- and adhesion molecule-functionalized, supported lipid bilayers, yielding a 2D Kd of ∼5,000 molecules/μm(2) This value is two to three orders of magnitude higher than previously measured 2D Kd values for interacting leukocyte surface proteins. Calculations indicated, however, that CD4/pMHC II binding would increase rates of T-cell receptor (TCR) complex phosphorylation by threefold via the recruitment of Lck, with only a small, 2-20% increase in the effective affinity of the TCR for pMHC II. The affinity of CD4/pMHC II therefore seems to be set at a value that increases T-cell sensitivity by enhancing phosphorylation, without compromising ligand discrimination.",

keywords = "Adhesion, Binding equilibrium and kinetics, Protein interactions, T-cell activation, TCR phosphorylation",

author = "Peter J{\"o}nsson and Southcombe, {Jennifer H} and Santos, {Ana Mafalda} and Jiandong Huo and Fernandes, {Ricardo A} and James McColl and Melissa Lever and Evans, {Edward J} and Alexander Hudson and Chang, {Veronica T} and Tom{\'a}{\v s} Hanke and Andrew Godkin and Dunne, {Paul D} and Horrocks, {Mathew H} and Matthieu Palayret and Screaton, {Gavin R} and Jan Petersen and Jamie Rossjohn and Lars Fugger and Omer Dushek and Xiao-Ning Xu and Davis, {Simon J} and David Klenerman",

year = "2016",

month = may,

day = "17",

doi = "10.1073/pnas.1513918113",

language = "English",

volume = "113",

pages = "5682--7",

journal = "Proceedings of the National Academy of Sciences",

issn = "0027-8424",

publisher = "The National Academy of Sciences of the United States of America",

number = "20",

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Jönsson, P, Southcombe, JH, Santos, AM, Huo, J, Fernandes, RA, McColl, J, Lever, M, Evans, EJ, Hudson, A, Chang, VT, Hanke, T, Godkin, A, Dunne, PD, Horrocks, MH, Palayret, M, Screaton, GR, Petersen, J, Rossjohn, J, Fugger, L, Dushek, O, Xu, X-N, Davis, SJ & Klenerman, D 2016, 'Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions', Proceedings of the National Academy of Sciences, bind 113, nr. 20, s. 5682-7. https://doi.org/10.1073/pnas.1513918113

Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions. / Jönsson, Peter; Southcombe, Jennifer H; Santos, Ana Mafalda et al.
I: Proceedings of the National Academy of Sciences, Bind 113, Nr. 20, 17.05.2016, s. 5682-7.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

TY - JOUR

T1 - Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions

AU - Jönsson, Peter

AU - Southcombe, Jennifer H

AU - Santos, Ana Mafalda

AU - Huo, Jiandong

AU - Fernandes, Ricardo A

AU - McColl, James

AU - Lever, Melissa

AU - Evans, Edward J

AU - Hudson, Alexander

AU - Chang, Veronica T

AU - Hanke, Tomáš

AU - Godkin, Andrew

AU - Dunne, Paul D

AU - Horrocks, Mathew H

AU - Palayret, Matthieu

AU - Screaton, Gavin R

AU - Petersen, Jan

AU - Rossjohn, Jamie

AU - Fugger, Lars

AU - Dushek, Omer

AU - Xu, Xiao-Ning

AU - Davis, Simon J

AU - Klenerman, David

PY - 2016/5/17

Y1 - 2016/5/17

N2 - The αβ T-cell coreceptor CD4 enhances immune responses more than 1 million-fold in some assays, and yet the affinity of CD4 for its ligand, peptide-major histocompatibility class II (pMHC II) on antigen-presenting cells, is so weak that it was previously unquantifiable. Here, we report that a soluble form of CD4 failed to bind detectably to pMHC II in surface plasmon resonance-based assays, establishing a new upper limit for the solution affinity at 2.5 mM. However, when presented multivalently on magnetic beads, soluble CD4 bound pMHC II-expressing B cells, confirming that it is active and allowing mapping of the native coreceptor binding site on pMHC II. Whereas binding was undetectable in solution, the affinity of the CD4/pMHC II interaction could be measured in 2D using CD4- and adhesion molecule-functionalized, supported lipid bilayers, yielding a 2D Kd of ∼5,000 molecules/μm(2) This value is two to three orders of magnitude higher than previously measured 2D Kd values for interacting leukocyte surface proteins. Calculations indicated, however, that CD4/pMHC II binding would increase rates of T-cell receptor (TCR) complex phosphorylation by threefold via the recruitment of Lck, with only a small, 2-20% increase in the effective affinity of the TCR for pMHC II. The affinity of CD4/pMHC II therefore seems to be set at a value that increases T-cell sensitivity by enhancing phosphorylation, without compromising ligand discrimination.

AB - The αβ T-cell coreceptor CD4 enhances immune responses more than 1 million-fold in some assays, and yet the affinity of CD4 for its ligand, peptide-major histocompatibility class II (pMHC II) on antigen-presenting cells, is so weak that it was previously unquantifiable. Here, we report that a soluble form of CD4 failed to bind detectably to pMHC II in surface plasmon resonance-based assays, establishing a new upper limit for the solution affinity at 2.5 mM. However, when presented multivalently on magnetic beads, soluble CD4 bound pMHC II-expressing B cells, confirming that it is active and allowing mapping of the native coreceptor binding site on pMHC II. Whereas binding was undetectable in solution, the affinity of the CD4/pMHC II interaction could be measured in 2D using CD4- and adhesion molecule-functionalized, supported lipid bilayers, yielding a 2D Kd of ∼5,000 molecules/μm(2) This value is two to three orders of magnitude higher than previously measured 2D Kd values for interacting leukocyte surface proteins. Calculations indicated, however, that CD4/pMHC II binding would increase rates of T-cell receptor (TCR) complex phosphorylation by threefold via the recruitment of Lck, with only a small, 2-20% increase in the effective affinity of the TCR for pMHC II. The affinity of CD4/pMHC II therefore seems to be set at a value that increases T-cell sensitivity by enhancing phosphorylation, without compromising ligand discrimination.

KW - Adhesion

KW - Binding equilibrium and kinetics

KW - Protein interactions

KW - T-cell activation

KW - TCR phosphorylation

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U2 - 10.1073/pnas.1513918113

DO - 10.1073/pnas.1513918113

M3 - Journal article

C2 - 27114505

SN - 0027-8424

VL - 113

SP - 5682

EP - 5687

JO - Proceedings of the National Academy of Sciences

JF - Proceedings of the National Academy of Sciences

IS - 20

ER -

Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions

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