Tissue specificity of cross-reactive allogeneic responses by EBV EBNA3A-specific memory T cells.
BACKGROUND
The crossreactivity of Epstein-Barr virus (EBV Epstein-Barr virus nuclear antigen 3A [EBNA3A])-specific CD8 T cells against allogeneic human leukocyte antigen (HLA)-B*44:02 has been shown to be dependent on presentation of self-peptide EEYLQAFTY by the target antigen. In this study, we report that allogeneic HLA-B*44:02 proximal tubular epithelial cells (PTECs) and human umbilical vein endothelial cells (HUVECs) are poor targets for EBV EBNA3A-specific T cells.


METHODS
The EEY peptide was exogenously loaded onto HLA-B*44:02 and HLA-B*44:03-expressing PTECs and HUVECs. EEY-peptide-loaded, and unloaded, PTECs and HUVECs were then incubated with serial dilutions of our EBNA3A T-cell clone, in a cytotoxicity assay.


RESULTS
Although HLA-B*44:02-expressing PTECs were specifically lysed in proportion to the effector/target ratio by the EBNA3A T-cell clone, without peptide loading, lysis was greatly increased by exogenous EEY peptide loading (15% vs. 75%; P<0.0001). HLA-B*44:02-expressing HUVECs were only lysed when loaded with exogenous EEY peptide (0% vs. 64%; P<0.0001). Lack of HLA expression and lack of ABCD3 gene expression were excluded as a cause for these results. PTECs and HUVECs were specifically targeted by another alloreactive T-cell clone without exogenous peptide loading, suggesting that the lack of recognition of HLA-B*44:02 epithelial and endothelial cells by the EBV EBNA3A T-cell clone was due to lack of EEYLQAFTY peptide presentation.


CONCLUSIONS
Tissue-specific (peptide dependent) alloreactivity may have important implications for transplantation monitoring and rejection.
Department of Immunohematology and Blood Transfusion, Leiden University Medical Centre, Leiden, The Netherlands. l.j.a.d_orsogna@lumc.nl
Lloyd J A
LJ
D'Orsogna
Dave L
DL
Roelen
Ellen M W
EM
van der Meer-Prins
Pieter
P
van der Pol
Marry E
ME
Franke-van Dijk
Michael
M
Eikmans
Jacqy
J
Anholts
Jamie
J
Rossjohn
James
J
McCluskey
Arend
A
Mulder
Cees
C
van Kooten
Ilias I N
II
Doxiadis
Frans H J
FH
Claas
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D002478:Cells, Cultured; D003429:Cross Reactions / Q000276:immunology; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology*; D015235:HLA-B Antigens / Q000378:metabolism; D059948:HLA-B44 Antigen; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000276:immunology; D009928:Organ Specificity / Q000276:immunology; D010455:Peptides / Q000378:metabolism
C063305:ABCD3 protein, human; D018528:ATP-Binding Cassette Transporters; C508151:EBNA-3A antigen; D019309:Epstein-Barr Virus Nuclear Antigens; D015235:HLA-B Antigens; D059948:HLA-B44 Antigen; D010455:Peptides