Production of a soluble gammadelta T-cell receptor to identify ligands for the murine intestinal intraepithelial gammadelta T cell population.
Although the functions and antigen recognition requirements of alphabeta T cells are well characterised, the antigens recognised by gammadelta T cells and the consequences of this recognition are unclear. gammadelta T cells are enriched within epithelia, where they eradicate transformed epithelial cells and regulate inflammation. To understand how this occurs, we need to understand the cellular ligands recognised by the gammadelta cell through the gammadelta T-cell receptor (TCR). We have therefore generated a soluble TCR (sTCR) to identify ligands for the murine gammadelta intestinal intraepithelial lymphocyte (IEL) population. sTCR was produced in the baculovirus expression system and purified by affinity chromatography on an anti-TCRdelta affinity column. sTCR was recognised by a panel of conformation-specific anti-TCRgammadelta antibodies. We will now use our sTCR to directly test the binding of putative ligands to the TCR using surface plasmon resonance, and to isolate the ligand biochemically.
Steele
Carrie R
CR
Department of Immunobiology, GKT School of Medicine, Guy's Hospital, SE1 9RT, London, UK.
Van Remoortere
Kier C
KC
Hayday
Adrian C
AC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002846:Chromatography, Affinity; D017931:DNA Primers; D018076:DNA, Complementary; D007413:Intestinal Mucosa / Q000166:cytology*; D008024:Ligands; D051379:Mice; D016133:Polymerase Chain Reaction; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000096:biosynthesis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D020349:Surface Plasmon Resonance
D017931:DNA Primers; D018076:DNA, Complementary; D008024:Ligands; D016692:Receptors, Antigen, T-Cell, gamma-delta; D011994:Recombinant Proteins