4. Concomitant therapy. Patients may receive concomitant medications to 
control the side effects of therapy (4,5). It is our plan to administer the 
same concomitant medications used in all previous TIL protocols. These Include 
acetaminophen (650 mg every 4 hours), indomethacin (50-75 mg every six hours) 
and ranitidine (150 mg every 12 hours) throughout the course of treatment. 
Patients may receive intravenous meperidine (25-50 mg) to control chills when 
they occur, although chills are unusual after the first one to two doses of 
IL-2. Hydroxyzine hydrochloride (25 mg every six hours) is given to treat 
pruritis. Steroids will not be used in these patients and if steroids are 
required, then the patient will Immediately be taken off protocol therapy. 
5. Preparation of the TNF-NeoR vector containing supernatant . The TNF- 
NeoR vector was constructed by modifying the Moloney murine leukemia vector 
by techniques similar to those previously described. Retroviral vector 
supernatant is produced by harvesting the cell culture medium from the PA317 
packaging line developed by Dr. A. Dusty Miller (22,23). This line has been 
extensively characterized and was used by us in our previous studies of the 
infusion of TIL modified by the LNL6 vector. The TNF-Neo vector preparations 
from PA317 will be extensively tested to assure that no detectable replication 
competent virus is present. Tests for replication competent virus will be 
conducted on both the vector supernatant and on the TIL after transduction. 
Testing will be the same as previously approved for the LNL6 supernatants used 
to introduce the NeoR gene into TIL (protocol 86-C-183c). The following tests 
will be run on the producer line and/or the viral supernatant: 
1) The viral titer will be determined on 3T3 cells. Viral preparations 
with titers greater than 5 x 10“^ colony forming units/ml will be used. 
2) Southern blots will be run on the producer line to detect the TNF 
gene. 
Recombinant DNA Research, Volume 14 
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