PRECLINICAL STUDIES USING IMMOBILIZED OKT3 TO ACTIVATE HUMAN T-CELLS FOR ADOPTIVE IMMUNOTHERAPY - OPTIMAL CONDITIONS FOR THE PROLIFERATION AND INDUCTION OF NON-MHC-RESTRICTED CYTOTOXICITY
Jp. Uberti et al., PRECLINICAL STUDIES USING IMMOBILIZED OKT3 TO ACTIVATE HUMAN T-CELLS FOR ADOPTIVE IMMUNOTHERAPY - OPTIMAL CONDITIONS FOR THE PROLIFERATION AND INDUCTION OF NON-MHC-RESTRICTED CYTOTOXICITY, Clinical immunology and immunopathology, 70(3), 1994, pp. 234-240
In order to obtain large numbers of T cells for adoptive immunotherapy
after bone marrow transplantation (BMT), we optimized conditions for
long-term proliferation of T cells that exhibit non-MHC-restricted cyt
otoxicity using immobilized anti-CD3 (OKT3) activation and culture in
IL-2. Proliferation and cytotoxicity directed at Daudi, K562, and B ce
ll lines were used to determine (1) the optimal concentration of IL-2
and the optimal time of exposure to immobilized OKT3 for maintaining g
rowth and cytotoxicity, (2) the starting populations that can be used,
(3) the T cell subsets that mediate cytotoxicity, and (4) the optimal
medium and concentration of serum for maintaining growth and cytotoxi
city. Peripheral blood lymphocytes (PBL) activated with OKT3 would pro
liferate and mediate cytotoxicity at IL-2 doses as low as 30 IU/ml. In
creasing the IL-2 concentrations beyond 600 IU/ml did not augment the
proliferative or cytotoxic responses of PBL. A 24-hr incubation on OKT
3 was sufficient to activate PBL. Increasing the incubation time on OK
T3 from 24 to 72 hr did not significantly enhance cytotoxicity. Compar
isons between PBL and purified T cells (E-rosette) indicated that eith
er cell population could be activated with OKT3 in the presence of IL-
2 to proliferate and mediate non-MHC-restricted cytotoxicity. Purified
populations of CD4(+) or CD8(+) T cells demonstrated equivalent proli
feration and cytotoxicity when activated using IL-2 and OKT3. With equ
al concentrations of human or fetal bovine serum, RPMI 1640 and X-Vivo
10 were comparable for supporting proliferation and cytotoxicity. The
se conditions are being used to activate and expand T cells for clinic
al trials that involve infusing activated T cells into recipients afte
r autologous BMT. (C) 1994 Academic Press, Inc.