Can oligomeric T-cell receptor be used as a tool to detect viral peptide epitopes on infected cells?
We have utilized soluble HIV Gag-specific T-cell receptor (TCR) D3 with low affinity and TCR-like antibody 25-D1.16 recognizing its natural peptide-MHC (pMHC) ligand with high affinity to determine how affinity and off-rate of the receptor-pMHC interactions affect the sensitivity of pMHC detection on the cell surface. We found that with soluble TCR cognate pMHCs can be detected only at relatively high cell surface densities when the TCR was oligomerized using either Streptavidin or quantum dot (QD) scaffolds. While the higher affinity probe led to a greater sensitivity of pMHC detection, monomers and oligomers of the probe showed essentially the same detection limit, which is restricted by the sensitivity of standard flow cytometry technique. We have also shown that imaging of QD/TCR specifically bound to cognate pMHC on the cell surface yielded a very bright fluorescent signal that can enhance the sensitivity of viral peptide detection on infected cells.
Department of Microbiology and Immunology and Kimmel Cancer Center, BLSB 650, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Nadia
N
Anikeeva
Tatiana
T
Mareeva
Wei
W
Liu
Yuri
Y
Sykulev
D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D002460:Cell Line; D004331:Drosophila melanogaster; D000939:Epitopes / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D012996:Solutions; D014764:Viral Proteins / Q000276:immunology*
D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D007140:Immunoglobulin Fab Fragments; D010455:Peptides; D011948:Receptors, Antigen, T-Cell; D012996:Solutions; D014764:Viral Proteins