ARTIFICIAL-CAPILLARY-SYSTEM DEVELOPMENT OF HUMAN ALLOREACTIVE CYTOTOXIC T-LYMPHOCYTES THAT LYSE BRAIN-TUMORS

The production of alloreactive cytotoxic T-lymphocytes (CTL) for thera py of recurrent brain tumours was performed in the CELLMAX(R) artifici al capillary system composed of cell-culture modules containing cellul ose acetate or cuprammonium rayon hollow fibres. Lymphocytes, obtained from the brain-tumour patient to be used for sensitization, were stim ulated with OKT3 (anti-CD3) and interleukin-2 (IL-2) and inoculated in to the extracapillary space of artificial capillary modules, For allog eneic CTL production, the expanded patient lymphocytes were harvested, irradiated and placed into a second artificial capillary system with allogeneic lymphocytes from a healthy donor, In a one-way mixed lympho cyte reaction, CTL developed in the presence of low-concentration IL-2 (60 i.u. of IL-2/ml), In 18-21 days the cell preparation usually disp layed a predominantly CD3+, CD8+ phenotype, which consorted with the d ual-labelled CD8/CD11a markers used to identify CTL, Chromium (Cr)-rel ease assays demonstrated lysis of patient tumour in relation to alloge neic glioma; the response observed in cold-target-inhibition assays co nfirmed lysis of the relevant tumour by the CTL.