Protocol ID93-008 
Page 3 
Expanded TIL were primarily CD3+CD4+TCRcxp+ (8 TIL-derived T-cell lines) and primarily 
CDS+CDS+TCRotP'*' in one TIL-derived T-cell line. Eleven patients (8 treated with TIL plus 
rIL-2 and 3 patients treated with rIL-2 alone) received a total of 38 cycles of rIL-2 without 
TIL. Grade 3 clinical toxicity (peritonitis) occurred in 1 of 9 four day TIL plus rIL-2 cycles 
and 1 of 38 four day cycles of rIL-2 alone. Grade 3 anemia occurred in 1 of 9 TIL plus rIL-2 
cycles and 3 of 38 cycles of rIL-2 alone. Two patients required blood transfusions. There 
were no measurable responses, however. 4 of 8 patients treated with IP TIL plus rIL-2 had 
some indication of clinical activity: ascites regression (2 patients), tumor and CA-125 
reduction (1 patient), and surgically confirmed stable tumor and CA-125 values (1 patient). 
The schedule of IP TIL plus low dose rIL-2 shows manageable toxicity and is worthy of 
further evaluation in patients with EOC who have less tumor burden. 
The most significant toxicities that have been described with IP adoptive cellular therapies 
are catheter related infections (25-28) and peritoneal fibrosis (25.27.28). These problems 
have been overcome by attention to the following details: (a) the use of lower doses and 
duration of IP rIL-2 such as we currently employ, i.e.. 6x10^ IU/m2/4 day cycle and. fb) 
restricting IP treatments to 3 cycles/course; and removal of IP catheters after 18 days; and 
(c) instituting prophylactic antibiotics at the time of IP catheter insertion. Using this 
approach we have observed no catheter related infections by performing microbiologic 
cultures on the tips of catheters removed from a number of patients after 18 days. 
2.6 Purification and expansion in rIL-2 of CD8 ~^ or CD4 '^ TIL-derived T-cell lines 
EbqDansion of ovarian TIL in rIL-2 may result in T-cell lines that are primarily CD3+CD8+CD4' 
apTGR+ or CD3+CD4+CD8'apTCR or mixtures of CD3+CD8+ and CD3+CD4+ T-cell lines. 
Purified CD4+ and CD8+ T-cell lines have been developed from rIL-2 expanded TIL using 
appropriate antibody coated flasks (CELLector devices. Applied Immune Sciences (AIS) Menlo 
Park. CA) (29). When rIL-2 expanded TIL reached 2x10® cells they are transferred in complete 
AIM V medium (includes rIL-2 at 600 lU/ml) to a CELLector-TIL sterile polystyrene chamber 
with anti-CD8 mAb covalently bound to the inner surface. CD8+ cells are separated from the 
mixture of CD4+ and CD8+ rIL-2-expanded TIL by positive selection. This device is further 
described in reference (29) and in a master drug file with the FDA. Within 2-4 days the cells 
detach from the matrix in the chamber and continue their expansion as described in our 
procedure for unseparated rIL-2 expanded T-cell lines (17). 
2.7 IP rIL-2 administration in combination with TIL. 
Ovarian TIL-derived T-cell lines require the presence in vitro of low concentrations of rIL-2 to 
sustain proliferation and for other functions including cytotoxicity (14.15.18) and in the 
production of cytokines (30). Since activated TIL derived T cells may require several days to 
concentrate at tumor sites (31) it is probably necessary to administer these cells with 
exogenous rIL-2 in order to maintain a suitable micro-environment for the cells after 
injection. Pharmacokinetic studies have shown that rIL-2 can be detected for a prolonged 
time after IP injection with an estimated half-life of 22 hrs following IP bolus injection (32). 
This compares with a half-life of only minutes when rIL-2 is administered by IV bolus 
injection. 
2.8 The use of neo^ marked cells to study trafficking of CD8 ~*' TIL-derived T cells in ovarian 
carcinoma patients who are receiving IP adoptive immunotherapy. 
We will determine whether purified CD3+CD4'CD8'^TCRaP'‘’ ovarian TIL-derived T cells marked 
with neo*^ that cire expanded in rIL-2 (600 lU/ml) for the companion treatment protocol ID92- 
015 (T92-0093) concentrate in metastatic tumor sites in the peritoneal cavity of patients with 
epithelial ovarian carcinoma. 
