Plaksin D, Polakova K, McPhie P, Margulies D H
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892, USA.
J Immunol. 1997 Mar 1;158(5):2218-27.
We have expressed in bacteria a single-chain T cell receptor (scTCR) with specificity for an HIV gp120-derived peptide bound to the murine MHC-I molecule, H-2Dd. This scTCR consists of V alpha covalently linked to the VbetaCbeta domains that was solubilized, refolded, and purified in high yield. Specific binding of the scTCR to MHC/peptide complexes was demonstrated by surface plasmon resonance, with a Kd of 2 to 8 x 10(-6) M. This scTCR specifically inhibited T cell activation, and stained cell surface MHC/peptide complexes as measured by cytofluorimetry. The preservation of binding specificity by such a three-domain scTCR suggests that this structure is sufficient for specific MHC/peptide recognition and that this strategy will be of general use as applied to other TCR.
我们已在细菌中表达了一种单链T细胞受体（scTCR），它对与小鼠MHC-I分子H-2Dd结合的HIV gp120衍生肽具有特异性。这种scTCR由与VβCβ结构域共价连接的Vα组成，该结构域经过溶解、重折叠并以高产率纯化。表面等离子体共振证明了scTCR与MHC/肽复合物的特异性结合，解离常数（Kd）为2至8×10⁻⁶ M。这种scTCR特异性抑制T细胞活化，并通过细胞荧光测定法检测到其对细胞表面MHC/肽复合物进行染色。这种三结构域scTCR对结合特异性的保留表明，该结构足以实现对MHC/肽的特异性识别，并且这种策略在应用于其他TCR时将具有普遍用途。
