Use of a Pan-Vβ primer permits the amplification and sequencing of TCRβ chains expressed by bovine T-cell clones following a single semi-nested PCR reaction

We report in this study the design and validation of a Pan-Vβ primer that in combination with Cβ-specific primers enables the amplification, in a single semi-nested PCR, of TCRβ chains expressed by bovine T-cell clones irrespective of the expressed Vβ sequence. Using the Pan-Vβ primer we examined the TCRβ chains expressed by 16 Theileria parva-specific clones that had not been previously analysed. TCRβ chain sequence was obtained from 15 of the clones following direct sequencing of the PCR product, whilst the other clone appeared to express 2 different TCRβ chains which were characterised following sub-cloning of the PCR product. We have also successfully used the Pan-Vβ primer to amplify the TCRβ chains expressed by 19 T-cell clones, on which previous analysis using Vβ-subfamily-specific primers had failed to do. Sequencing of these TCRβ chains has identified members of 2 novel bovine Vβ subfamilies—Vβ5 and VβX. This method offers a simple and rapid method of analyzing the TCRβ chains of bovine T-cell clones that has many potential applications in the investigation of bovine T-cell responses.