Donor material
Stocks of donor leukapheresis mononuclear cells, derived EBV-specific T
cells and corresponding LCL from the SNBTS second-generation bank were
supplied from frozen stored stocks. For this bank material, initial
leukapheresis donations were collected by Spectra Optia apheresis system
(Terumo BCT) and all donations were fully consented for research use and
were either from New Zealand (NZ code, free of variant CJD and suitable
for therapy); or were from local Scottish donors, for research only.
Autologous EBV-transformed lymphoblastoid cell lines (LCL) for the
second-generation bank were generated from NZ donor PBMC by infection
with concentrated EBV-positive supernatant, for use as autologous
antigen presenting cells as previously described [12]. Briefly,
supernatant from a live EBV-expressing cell line was added to donor PBMC
and cultured for several rounds until stable LCL lines were established.
The LCL were frozen in 1ml aliquots in human serum albumin (HSA) + 10%
DMSO until use. Initial work on phenotypic development was run using
peripheral blood mononuclear cells (PBMC) derived from buffy coats from
normal blood donors, supplied through SNBTS under Sample Governance
14~11.