Functional CD4(+) and CD8(+) T-cell responses induced by autologous mitomycin C treated Epstein-Barr virus transformed lymphoblastoid cell lines

Epstein-Barr virus (EBV) gene expression in tumor cells of posttransplant l ymphoproliferative disorder (PTLD) patients resembles that of EBV transform ed B-cell lines (LCL). EBV-specific cytotoxic T-lymphocytes can be generate d by stimulating peripheral blood lymphocytes with autologous LCL. We descr ibe a standardized method for the growth inactivation and cryopreservation of LCL for optimal T-cell stimulation and analyzed the function and phenoty pe of responding T-cells. LCL growth was completely blocked by mitomycin C treatment (McLCL) and McLCL could be cryopreserved while retaining excellen t APC function. McLCL stimulated both CD4(+) and CD8(+) T-cells as measured by HLA-DR and CD25 expression using FACS analysis. EBV-specific CTL activi ty and T-cell proliferation were induced and immunocytochemical staining sh owed CD4(+) and (granzyme B positive) CD8(+) T-cells resetting with McLCL. Granzymes A and B, IFN-gamma, and IL-6 were detected at significant levels in the supernatant. Thus, ex vivo T-cell activation with cryopreserved McLC L results in activation of both CD4(+) and CD8(+) T-cells producing a Th1-l ike cytokine profile, making this a suitable protocol for adoptive therapy of PTLD. (C) 2001 Academic Press.