presence of remaining tumor. At least 200 cells are studied and therapy 
proceeds only when no tumor cells are found. Other lymphocyte samples are 
taken for characterization of cell surface markers and for assessment of 
cytotoxicity using techniques identical to that in our previous protocol (86- 
C-183; reference 23, attached). Briefly, lymphocytes are stained with 
fluorescent-labeled antibodies (Leu2, Leu3, Leu4, Leu7, Leull, Leu5, Leu9, 
LeuM3 , HLADDR and Tac). Chromium release assays are performed with K562, 
Daudi autologous tumor, and allogeneic tumor targets. 
To infuse the lymphocytes they will be thawed and grown for one to three 
additional' weeks using the same procedures detailed above. For infusion TIL 
are reharvested. At the time of cell collection, one liter of saline for 
injection is pumped through the collection chamber and the centrifuge is 
stopped. Lymphocytes are resuspended in the collection bag, the centrifuge is 
started again, and another liter of saline is pumped through to fully wash the 
free of tissue culture medium components. The cells are then filtered through 
a platelet administration set into 600 ml transfer packs (Fenwal), and 50 ml 
of 25% albumin and 450,000 IU of IL-2 are added to the 200 to 300 ml volume of 
cells in saline. The TIL are infused over 30 to 60 minutes through a central 
venous catheter. 
7. Interleukin-2 : The recombinant IL-2 used in this trial will be 
provided by the Division of Cancer Treatment, National Cancer Institute 
(supplied by the Cetus Corporation, Emeryville, CA) (28) and will be 
administered exactly as specified in our previously approved protocols (86-C- 
183c) (41). The IL-2 will be provided as a lyophilized powder and will be 
reconstituted with 1.2 ml/vial. Each vial contains approximately 1.2 mg of 
IL-2 (specific activity 18 x 10 6 IV/mg) . Less than 0.04 ng of endotoxin are 
present per vial as measured by the limulus amebocyte assay. Each vial also 
contains 5% mannitol and approximately 130-230 ug of sodium dodecyl sulfate/mg 
of IL-2. Following reconstitu tion the IL-2 will be diluted in 50 ml of normal 
saline containing 5% human serum albumin and will be infused intravenously at 
a dose of 720,000 IU/kg over a 15 minute period every 8 hr, beginning from two 
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Recombinant DNA Research, Volume 15 
