| First Author | Zhao J | Year | 2013 |
| Journal | J Immunol Methods | Volume | 396 |
| Issue | 1-2 | Pages | 56-64 |
| PubMed ID | 23928495 | Mgi Jnum | J:287216 |
| Mgi Id | MGI:6403656 | Doi | 10.1016/j.jim.2013.07.011 |
| Citation | Zhao J, et al. (2013) Development of transgenic mice expressing a coronavirus-specific public CD4 T cell receptor. J Immunol Methods 396(1-2):56-64 |
| abstractText | Mice that are transgenic (Tg) for T cell receptor (TCR) expression are used extensively to analyze longitudinal T cell responses during effector and memory phases of the T cell response. Generation of TCR Tg mice generally requires T cell stimulation and cloning in vitro prior to amplification, processes which introduce biases into selection of the TCR that is ultimately chosen for TCR Tg mouse generation. Here we describe an alternative approach that involves no T cell stimulation or propagation in vitro. We generated mice that were transgenic for a TCR responding to a CD4 T cell epitope (epitope M133) that is immunodominant in mice infected with a neurotropic coronavirus, the JHM strain of mouse hepatitis virus. The CD4 T cell response to epitope M133 is of particular interest because it may be pathogenic, protective or regulatory, depending upon the physiological setting. We applied an iterative process in which we identified a TCR-beta chain expressed by all mice that were examined ('public sequence'). This TCR-beta chain was introduced into bone marrow cells with a lentivirus vector, generating TCR-beta retrogenic mice. A TCR-alpha chain that paired with this TCR-beta was then identified and used to generate a second set of TCR (alpha/beta) retrogenic mice. After demonstrating that these cells were functional and responded to epitope M133, these TCR chains were used to generate an epitope M133-specific TCR Tg mouse. This method should be generally useful for engineering TCR Tg mice without introduction of bias caused by in vitro manipulation and propagation. |
