5) PCR assay for the absence of 4070A envelope gene must be negative. 
6) Reverse transcriptase assay must be negative. 
7) Southern blot will be run on the transduced TIL to assure that Intact 
provirus is present. 
8) TNF protein assay to assure the production of TNF. Cells must be 
producing at least 150 pg TNF/10^ cells/24 hours. 
9) IL-2 will be withdrawn from the culture medium of an aliquot for at 
least one week to assure that cells do not exhibit autonomous growth in the 
absence of IL-2. 
10) Cytotoxicity against the autologous and at least two other targets 
will be tested. The phenotype of the cells will be tested by fluorescence 
activated cell sorting analysis. 
The S+/L- assay, the 3T3 amplification assay and the polymerase chain 
reaction assay are detailed in Appendix C. 
Recombinant DNA Research, Volume 14 
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