MHC-independent αβ T cells replace epidermal and dermal γδ T cells in Tcrd-/- mice

In Tcrd-/- mice, dendritic epidermal T cells (DETC) are known to be replaced by a DETC-population expressing αβ TCR. Here, the TCR-repertoire from Tcrd-/- mouse skin T cells was compared to the one from WT mice. The analysis revealed the existence of T cells expressing MHC-independent αβ TCR and replacing the missing γδ T cells in Tcrd-/- mouse epidermis and dermis. For TCR-sequencing the desired populations were sorted from WT or Tcrd-/- mice by flow cytometry as living CD45+CD3+TCRβ+ (WT dermal, Tcrd-/- dermal, Tcrd-/- pLN), living CD45+CD3hiTCRβhi (Tcrd-/- DETC), living CD45+CD3hiTCRγδhi (WT DETC) and living CD45+CD3+TCRβ+CD103+ or CD103hi (Tcrd-/- dermal CD103+, Tcrd-/- dermal CD103hi). Each sample is a pool from equivalent populations from several (5 to 18) animals. RNA was isolated from these cells and was analyzed by 5´RACE and PCR amplification followed by illumina-sequencing of Tcrb or Tcrg transcripts.