Making a stabilized HLA-E complex with native properties.

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B918
Abstract ID: 5409

Presenting Author:
Thomas H Blicher , Senior Scientist at Immudex

Abstract:

Objective

The non-classical, human MHC-I molecule HLA-E is functionally monomorphic and therefore a highly attractive target in immunotherapy against cancers and infectious diseases. However, HLA-E is characterized by low stability due to its biological role as a rapid-response system to monitor HLA-I expression. The inherent instability makes production of HLA-E difficult and severely limits its use as a reagent, which in turn negatively impacts the understanding of HLA-E biology. Here, we present native-like, stabilized forms of HLA-E, which brings production of HLA-E in line with that of classical MHC-I molecules.

 

Methods

Disulfide bridges were introduced in HLA-E at selected locations deemed to not interfere with recognition of HLA-E by NK or T cell receptors. The stability of the modified HLA-E molecules was monitored using thermal denaturation, and conservation of native HLA-E properties was assessed by in vitro TCR-binding assays and through staining of NK cells.

 

Results

The modified HLA-E molecules exhibited increased stability, increased refolding yields, both or neither. The most stable HLA-E variant exhibited thermal stability comparable to complexes of HLA-A*02:01, while retaining native-like recognition by both NK and T cell receptors.

 

Summary

Stabilized forms of HLA-E have been generated and their native-like properties validated using in vitro assays and NK cell staining. This enables production of reagents for the study HLA-E biology.