RT Journal Article
SR Electronic
T1 Multiple sclerosis
JF Neurology - Neuroimmunology Neuroinflammation
JO Neurol Neuroimmunol Neuroinflamm
FD Lippincott Williams & Wilkins
SP e241
DO 10.1212/NXI.0000000000000241
VO 3
IS 4
A1 Rühl, Geraldine
A1 Niedl, Anna G.
A1 Patronov, Atanas
A1 Siewert, Katherina
A1 Pinkert, Stefan
A1 Kalemanov, Maria
A1 Friese, Manuel A.
A1 Attfield, Kathrine E.
A1 Antes, Iris
A1 Hohlfeld, Reinhard
A1 Dornmair, Klaus
YR 2016
UL http://nn.neurology.org/content/3/4/e241.abstract
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