Expression of two distinct T cell receptors by EL-4 T lymphoma cells

Objective:To investigate the expression of T cell receptor(TCR) inEL-4 T lymphoma cells.Methods:Mouse spleen cells were used as positive control,total RNA was isolated from mouse spleen cells and EL-4 cell line.Complementarity Determining Region 3(CDR3) of 24 TCR BV gene subfamilies was amplified by the reverse transcription polymerase chain reaction(RT-PCR).To determine the spectratypes and sequences of CDR3 region of TCR BV subfamilies expressed by EL-4,PCR products were analyzed by TCR GeneScan Technique and DNA sequencer,respectively.Results:24 TCR BV subfamilies of mouse spleen cells were polyclonal,the spectratypes of TCR CDR3 region were showed Gaussian distribution.EL-4 was shown detected the expression of TCR BV10 and BV12 simultaneously,spectratypes of which showed single peak,and the TCR sequences of BV10 and BV12 were confirmed by DNA sequencer and they were both in-frame rearrangements.Conclusion:Two sets of in-frame rearrangements of TCR β chain exist in EL-4,and there maybe 'defects' in allelic exclusion in TCR β chain.It provides a basis for future research of allelic exclusion mechanism.