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. 2011 Jan 8;1(12):1118–1127. doi: 10.1007/s13238-010-0144-5

High affinity soluble ILT2 receptor: a potent inhibitor of CD8+ T cell activation

Ruth K Moysey 1, Yi Li 1, Samantha J Paston 1, Emma E Baston 1, Malkit S Sami 1, Brian J Cameron 1, Jessie Gavarret 1, Penio Todorov 1, Annelise Vuidepot 1, Steven M Dunn 1, Nicholas J Pumphrey 1, Katherine J Adams 1, Fang Yuan 1, Rebecca E Dennis 1, Deborah H Sutton 1, Andy D Johnson 1, Joanna E Brewer 1, Rebecca Ashfield 1, Nikolai M Lissin 1, Bent K Jakobsen 1,✉
PMCID: PMC4875079  PMID: 21213105

Abstract

Using directed mutagenesis and phage display on a soluble fragment of the human immunoglobulin super-family receptor ILT2 (synonyms: LIR1, MIR7, CD85j), we have selected a range of mutants with binding affinities enhanced by up to 168,000-fold towards the conserved region of major histocompatibility complex (MHC) class I molecules. Produced in a dimeric form, either by chemical cross-linking with bivalent polyethylene glycol (PEG) derivatives or as a genetic fusion with human IgG Fc-fragment, the mutants exhibited a further increase in ligand-binding strength due to the avidity effect, with resident half-times (t 1/2) on the surface of MHC I-positive cells of many hours. The novel compounds antagonized the interaction of CD8 co-receptor with MHC I in vitro without affecting the peptide-specific binding of T-cell receptors (TCRs). In both cytokine-release assays and cell-killing experiments the engineered receptors inhibited the activation of CD8+ cytotoxic T lymphocytes (CTLs) in the presence of their target cells, with subnanomolar potency and in a dose-dependent manner. As a selective inhibitor of CD8+ CTL responses, the engineered high affinity ILT2 receptor presents a new tool for studying the activation mechanism of different subsets of CTLs and could have potential for the development of novel autoimmunity therapies.

Keywords: CD8+ T cells, cellular activation, autoimmunity, cell surface molecules, binding affinity, phage display

Footnotes

These authors contributed equally to this work.

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