Generation of stimulated, lymphokine activated T killer (T-LAK) cells from the peripheral blood of normal donors and adult patients with recurrent glioblastoma

Therapy of recurrent high grade gliomas with surgery, and autologous mitogen activated IL-2 stimulated killer (MAK) Lymphocytes: I. Enhancement of MAK lytic activity and cytokine production by PHA and clinical use of PHA

“…These activated T cells have been given various names: 1) PHA Activated Killer (PAK) Cells [34,35], 2) Mitogen Activated IL-2 Stimulated Killer (MAK) Cells, 3) Autologous Stimulated Lymphocytes (ASL) [33], and CD3 or T-LAK [4,5].…”

“…Approximately 10 9 cells were collected from leukapheresis of 1-2.5 liters of blood. These cells were stimulated for 48 hours with PHA and cultured in IL-2 (RO-23-6019) for approximately 10 days, as described initially by Ingram et al [19] and further characterized by Yamamoto et al [4]. A second leukapheresis was done 3-4 days before surgery to obtain 109 cells.…”

“…Cytolytic assays employing non-adherent target cells K562 are performed in 96 well round-bottomed assay plates, whereas cytolytic assays for adherent U373 MG cells are performed in 96 well flat-bottomed assay plates as described by Yamamoto et al [4]. Radiolabeling of the non-adherent target cells is accomplished by the addition of 20 ul of 51Cr with a specific activity of 183 mCi/mg (ICN, Irvine, CA), at a concentration of 5 mCi/ml to 106 cells in 1 ml of serum free media for 1-2 hours at 37 ~ C. After 1-2 hours at 37 ~ C, the cells are washed 3 times (300 x G for 5 min) with cold RPMI-10% FBS.…”

“…There are several types of lymphokine-activated effector cells: A) Natural Killer (NK)-like cells derived lymphokine-activated killer (LAK) cells [2,3], B) non-MHC restricted T killers (mitogen activated killer (MAK) cells [1,[3][4][5], and C) tumor infiltrating lymphocytes (TIL's) [6,7]. These activated lymphocytes when administered systemically along with IL-2 have been employed to treat metastatic melanoma, sarcomas, adenocarcinomas, and implanted gliomas, melanoma, and sarcomas in animal models.…”

“…Lymphocytes can be activated in vitro with Interleukin-2 (IL-2) to produce cells that can kill tumor cells in a non-major histocompatibility restricted (MHC) manner, and secrete lymphokines in vitro [1][2][3][4]. There are several types of lymphokine-activated effector cells: A) Natural Killer (NK)-like cells derived lymphokine-activated killer (LAK) cells [2,3], B) non-MHC restricted T killers (mitogen activated killer (MAK) cells [1,[3][4][5], and C) tumor infiltrating lymphocytes (TIL's) [6,7].…”