PT  - JOURNAL ARTICLE
AU  - Rühl, Geraldine
AU  - Niedl, Anna G.
AU  - Patronov, Atanas
AU  - Siewert, Katherina
AU  - Pinkert, Stefan
AU  - Kalemanov, Maria
AU  - Friese, Manuel A.
AU  - Attfield, Kathrine E.
AU  - Antes, Iris
AU  - Hohlfeld, Reinhard
AU  - Dornmair, Klaus
TI  - Multiple sclerosis
AID  - 10.1212/NXI.0000000000000241
DP  - 2016 Aug 01
TA  - Neurology - Neuroimmunology Neuroinflammation
PG  - e241
VI  - 3
IP  - 4
4099  - http://nn.neurology.org/content/3/4/e241.short
4100  - http://nn.neurology.org/content/3/4/e241.full
SO  - Neurol Neuroimmunol Neuroinflamm2016 Aug 01; 3
AB  - Objective: To identify target antigens presented by human leukocyte antigen (HLA)–A*02:01 to the myelin-reactive human T-cell receptor (TCR) 2D1, which was originally isolated from a CD8+ T-cell clone recognizing proteolipid protein (PLP) in the context of HLA-A*03:01, we employed a new antigen search technology.Methods: We used our recently developed antigen search technology that employs plasmid-encoded combinatorial peptide libraries and a highly sensitive single cell detection system to identify endogenous candidate peptides of mice and human origin. We validated candidate antigens by independent T-cell assays using synthetic peptides and refolded HLA:peptide complexes. A molecular model of HLA-A*02:01:peptide complexes was obtained by molecular dynamics simulations.Results: We identified one peptide from glycerolphosphatidylcholine phosphodiesterase 1, which is identical in mice and humans and originates from a protein that is expressed in many cell types. When bound to HLA-A*02:01, this peptide cross-stimulates the PLP-reactive HLA-A3-restricted TCR 2D1. Investigation of molecular details revealed that the peptide length plays a crucial role in its capacity to bind HLA-A*02:01 and to activate TCR 2D1. Molecular modeling illustrated the 3D structures of activating HLA:peptide complexes.Conclusions: Our results show that our antigen search technology allows us to identify new candidate antigens of a presumably pathogenic, autoreactive, human CD8+ T-cell-derived TCR. They further illustrate how this TCR, which recognizes a myelin peptide bound to HLA-A*03:01, may cross-react with an unrelated peptide presented by the protective HLA class I allele HLA-A*02:01.58-2D1-CD8-sGFP=58α−β− T hybridoma cells expressing TCR 2D1, human CD8αβ molecules, and sGFP under the control of nuclear factor of activated T cells; 58-B7-CD8-sGFP=58α−β− T hybridoma cells expressing TCR B7, human CD8αβ molecules, and sGFP under the control of nuclear factor of activated T cells; APC=antigen-presenting cells; COS-7-A2=COS-7 cells expressing HLA-A*02:01; COS-7-A3=COS-7 cells expressing HLA-A*03:01; DMXL2=Dmx-like-2; EAE=experimental autoimmune encephalomyelitis; EML5=echinoderm microtubule associated protein-like 5; GPCPD1=glycerolphosphatidylcholine phosphodiesterase 1; HLA=human leukocyte antigen; HLA-A2=HLA-A*02:01; HLA-A3=HLA-A*03:01; IL=interleukin; Met=methionine; MHC=major histocompatibility complex; MS=multiple sclerosis; mTECs=medullary thymic epithelial cells; NCAN=Neurocan; NFAT=nuclear factor of activated T cells; PECP=plasmid-encoded combinatorial peptide; PLP=proteolipid protein; RMSD=root mean square deviation; PLP=proteolipid protein; TAX=T-lymphotrophic virus-2 protein; TCR=T-cell receptor; VMD=visual molecular dynamics