reconstituted in volumes of 50 to 500 ml of 5% dextrose solution with 0.1% human albumin. It 
is recommended that in-line filters not be used for delivery of ProleukinR since bioassays 
have shown significant loss of IL-2 activity after filtration. Reconstitution with sterile water or 
normal saline solutions for intravenous injection should be avoided because of precipitation. 
In addition, ProleukinR should not be mixed with other drugs. The lyophilized material is 
stable at 2-8°C for 18 months from the date of manufacture. This material should be 
protected from excessive exposure to light during extended storage. 
4.2 QKT3 
OKT 3 is a murine monoclonal antibody (mAb) to the CD3 antigen on human T cells. It 
is manufactured by Ortho Pharmaceutical Corp. and will be supplied by CTEP, NCI. It has 
been used extensively as an intravenously administered immunosuppressive agent in organ 
transplant patients. The mAb is a biochemically purified lgG2a immunoglobulin. It is 
supplied in a 5 ml ampule (Ortho clone OKT 3 R ) containing 5 mg (1 mg/ml) of mAb in a 
sterile, colorless solution which may contain a few fine translucent protein particles. Each 
ampule contains a buffered solution (pH 7.0 ± 0.5) of monobasic sodium phosphate (2.25 
mg), dibasic sodium phosphate (9.0 mg), sodium chloride (43 mg) and polysorbate 80 (1.0 
mg) in water. 
4.3 Retroviral vector: GBAH4-8 
GBAH4-18 is a Moloney Murine Leukemia virus vector which encodes for human IL-4. 
The retrovirus producer cell line is currently undergoing safety testing as is required by the 
FDA. Once testing is complete , then a 1-2 liter lot of virus stock will be 
prepared by an FDA-approved commercial facility and subjected to tests for 
replication competent virus , bacterial, viral, and fungal agents as is required 
clinical grade material produced by cell lines. 
4. 3. a. CQn§taictiQn-Qlilisj£lr.Q.Yiru s ve ctor G BAH4-18 - 
The parental retrovirus vector used for these studies is the gag BA, which has 
been extensively studied, and was the parental vector for a previous human gene therapy 
protocol approved by the RAC (#9110-12). The general structure of the vector will be 
described, using the published numbering of the provirus sequences (24). The backbone 
structure of this vector includes an intact 5' LTR of Moloney Murine Leukemia Virus (Mo- 
MLV), with additional Mo-MLV sequences between the 5* LTR and the internal promoter 
spanning nucleotide 146 at the border of U5 to the natural Xho 1 site in the gag coding 
region at nucleotide 1560 (with the exception that a Sac II linker was inserted at the Hae III 
site at nucleotide 624 (25). The plasmid also contains wild-type Mo-MLV sequences from the 
Cla I site at nucleotide 7674 (which was converted to a Bam HI site with synthetic linkers) to 
the end of the 3' LTR. Sequences containing the viral enhancer elements of the 3' LTR from 
the Pvu II site at nucleotide 7933 to the Xba site at nucleotide 81 1 1 have been deleted. In 
addition to these sequences, there are flanking mouse genomic DNA and pBR322 
sequences (spanning the Hind III site to the Eco R 1 site). The internal promoter used in this 
vector was derived from a Xho 1 to Mbo 1 fragment of the chicken p-actin gene spanning 
nucleotides -266 to +1(26). The Mbo 1 site was converted to a Bam HI site and the modified 
p-actin fragment was cloned into the parental vector. The plasmid pCDhlL-4, containing the 
complete human IL-4 cDNA, was digested with Mse 1 , and a 472 base pair fragment 
containing base pairs 61-513 of the human IL-4 cDNA sequence was obtained (27). The 
fragment was blunt ended with Klenow and deoxynucleotides, synthetic Bam HI linkers 
Recombinant DNA Research, Volume 18 
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