Loss of Original Antigenic Specificity in T Cell Hybridomas Transduced with a Chimeric Receptor Containing Single-Chain Fv of an Anti-Collagen Antibody and FcεRI-Signaling γ Subunit

This is the authors' abstract. We don't add key points for this paper.

T cell hybridomas HCQ6 and MD.45 acquired Ab-type specificity to collagen type II, when engrafted with a chimeric cell surface receptor, scC2Fv/γ, which includes the single-chain Fv domain (scFv) of the anti-collagen type II mAb C2 and the signaling γ subunit of the FcεRI. When transduced into MD.45 cells, scC2Fv/γ or its mutated form lacking immunoreceptor tyrosine-based activation motif (ITAM), scC2Fv/γIC−, formed mainly homodimers. A small proportion of these molecules formed heterodimers with endogenous CD3ζ in these hybridoma cells. By contrast, in HCQ6 cells, the majority of scC2Fv/γ and scC2Fv/γIC− molecules formed heterodimers with CD3ζ, and only a small proportion of them was expressed as homodimers. Stimulation with plastic-immobilized collagen induced IL-2 production in scC2Fv/γ-transduced MD.45 cells, but not in MD.45 cells transduced with the ITAM-less chimera scC2Fv/γIC−. HCQ6 cells transduced with scC2Fv/γ responded to plastic-bound collagen. Due to the high content of CD3ζ-associated chimeras, HCQ6 cells transduced with the ITAM-less scC2Fv/γIC− chimera were also responsive to plastic-bound collagen. When cells were stimulated with collagen in solution, MD.45 cells transduced with scC2Fv/γ produced IL-2, whereas transduced HCQ6 cells were unresponsive, hence suggesting that the ability of cells transduced with scC2Fv chimeras to respond to soluble collagen correlated with predominant expression of divalent scC2Fv/γ homodimers, but not monovalent scC2Fv/γ-CD3ζ or scC2Fv/γIC−-CD3ζ heterodimers. Of interest, expression of CD3 subunits in hybridomas transduced with scC2Fv chimeras was reduced, resulting in decreased response to cognate Ags.