HLA-D typing using Epstein-Barr virus-induced lymphoblastoid cell lines.

We examined whether Epstein-Barr virus-induced lymphoblastoid cell lines, established from homozygous typing cells (HTC-EBV-LCL), could be substituted for normal homozygous typing cells (HTC) in HLA-D typing. When using EBV-LCL as stimulators in one-way MLR, the stimulator: responder ratio was found to be most important. At a ratio of 1:10, the autologous MLR between EBV-LCL and PBL from the same donors exhibited low double normalized values (DNV) which could be distinguished from those of homologous combinations. The panel cells typed with HTC always exhibited low DNV in repeat one-way MLR in which EBV-LCL established from the same HTC were used. On the other hand, by employing our previously described method, we were able to establish the necessary HTC-EBV-LCL from unselected seropositive adult donors and utilize these cells for HLA-D typing. We suggest that our method makes it possible to circumvent the problem presented by the shortage of HTC, since our HTC-EBV-LCL can be substituted for normal HTC in HLA-D typing.