Protocol ID93-008 
Page 6 
smaller number of patients being required. DNA will be extracted by a proteinase K phenol 
procedure and the neo^ gene quantitated by PCR to determine the fold increase at metastatic 
tumor sites. The proportions of CD3+CD8+ T cells will be determined in biopsies utilizing 
OKT8 and OKT3 moAbs by histochemical procedures that we have established (44). In 
addition to these objectives, we will also determine the persistence of neo^ marked T cells in 
the abdominal cavity during the course of treatment with TIL + rlL-2. 
BACKGROUND DRUG INFORMATION AND PREPARATION OF NEO^ TRANSDUCED CELLS 
3. 1 Proleukin recombinant interleukin-2 a (human) (Chiron-CETUS) 
A clone containing the rIL-2 gene was derived from ds cDNA synthesized from mRNA 
extracted from the human Jurkat cell line. The rIL-2 protein is produced by 
Escherichia coli containing the pBR322 plasmid into which the gene was inserted. 
Proleukin differs from native rIL-2 in that it is not glycosylated and has no N-terminal 
alanine, and a serine substituted for cystine at the amino acid position 125. Its 
molecular weight is approximately 15,600 daltons. 
3.1.1 Reconstitution 
Proleukin is supplied as a lyophilized cake in 5 cc vials containing 1 mg protein. 
Reconstitution Directions; IL-2 (Chiron), NSC #373364, is provided as a lyophilized 
powder. Each 5 cc vial has a labeled strength of 1.3 mg (22 million lU). The vial is 
reconstituted with 1.2 mL of Sterile Water for Injection, USP, and the resultant 
concentration is 18 million lU/ml. Direct the diluent against the side of the vial to 
avoid excess foaming. Swirl contents gently until completely dissolved. Do not shake. 
The resulting solution should be a clear, colorless liquid. 
Reconstituted Proleukin is suitable for intravenous injection directly or may be diluted 
as necessary in volumes of 50 mL to 500 mL with 5% Dextrose injection, USP with 
0.1% Albumin Human, USP. When diluting, the Albumin Human, USP should be 
added to the 5% Dextrose injection, USP prior to the addition of the reconstituted 
Proleukin. 
For TIL administration dilution of original reconstituted rIL-2 is performed in 5% 
dextrose in 0.2% Normal Saline with 0. 1% Albumin Human, USP. 
Reconstituted and diluted Proleukin contain no bacteriostatic agents. Sterile Technique 
should be carefully observed during their preparation and administration. 
3.1.2 Stability 
Lyophilized Cake In Vial. Stable at 2-8°C for 18 months from date of manufacture. 
Protect the lyophilized material during extended storage from excessive exposure to 
light. 
Proleukin, diluted for intravenous administration in 5% Dextrose injection, USP, in a 
plastic bag (e.g. Viaflex. manufactured by Travenol Laboratories. Inc.) containing 0.1% 
Albumin Human, is stable for 48 hours at refrigerated and room temperature 2° to 30°C 
(36° to 86° F). 
3.1.3 Suppliers 
All rIL-2 for this project will be supplied by the National Cancer Institute. CELLector T- 
150 Cell Selection Procedure device will be supplied by Applied Immune Sciences Inc., 
Menlo Park. CA. 
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Recombinant DNA Research, Volume 19 
