Protocol ID93-008 
Page 4 
2.8.1 The treatment protocol. 
The purified 008"^ T cells will be expanded to large numbers (10'° to 10** range) in low 
concentrations of rlL-2 (600 lU/ml) by a four-step method that we have developed (18). 
Ovarian cancer patients who have failed at least first-line chemotherapy will be eligible to 
receive IP injections of rIL-2 expanded T cells together with low doses of rIL-2 under the 
companion treatment protocol ID92-015 (T92-0093). This protocol is approved by the Cancer 
Therapy Evaluation Program at the National Cancer Institute and by the M. D. Anderson 
Surveillance Committee (IRB). Under this protocol, patients will undergo as part of their 
clinical evaluation: (1) serial sampling of cells from the peritoneal catheter used for 
administering TIL + rlL-2 into the peritoneal cavity, and (2) laparoscopic evaluation for 
clinical response one month after IP treatment with CD8+ T cells. 
Metastatic lesions of ovarian cancer are usually observed as surface lesions on the 
peritoneum of the abdomino-pelvic cavity and on the serosa of the intestines. These surface 
metastases shed tumor cells and TIL into the abdominal cavity which are easily retrieved for 
growing TlL-derived T-cell lines as we have shown in previous experiments (14-17). The 
situation of these metastases are monitored by video laparoscopy. The metastatic lesions of 
ovarian cancer are in direct contact with TlL-derived T cells utilized in IP adoptive 
immunotherapy. Only the patients who are enrolled on the above treatment protocol will be 
eligible to have their rIL-2 expanded TIL marked with the neo*^ gene as described in this 
protocol. 
2.8.2 Rationale for using neo *^ transduced cells in vivo. 
CD8+ TIL derived T cells may exhibit a therapeutic effect in vivo by a number of different 
mechanisms. These TIL may have therapeutic effects either directly by cell to cell contact or 
indirectly through the production of a number of cytokines such as interferon-yTNFa or 
GMCSF. Our studies have shown that CD8'*’ ovarian T cells may exhibit HLA class I 
restricted autologous tumor specificity (14, 15) or may produce cytokines in a class I restricted 
manner in response to autologous tumor cells (30). These cytokines may exhibit antitumor 
activity by one or more of the following mechanism(s): (a) by direct lysis of ovarian tumor 
cells: (b) by enhancing HLA Class I and II expression on tumor cells that can now be 
recognized more easily by MHC restricted T cells; (c) by inducing the differentiation of pre- 
effector T cells to effector T cells. The objectives of this protocol are to determine whether the 
in vitro expanded purified CD8+ T cells home to the tumor sites. Data from these experiments 
will enhance our understanding of how these cells work in vivo and permit the development of 
more effective adoptive immunotherapeutic strategies for patients with ovarian cancer. 
Previous clinical experiments in melanoma conducted with TIL labeled with * * * Indium oxlne 
have provided very limited Information on TIL trafficking, largely because of the short half-life 
of **Mndium and because ***Indium is preferentially taken up by the liver (31). For these 
reasons we were encouraged by experiments utilizing retroviral vectors for marking of TIL that 
were conducted by Rosenberg and associates (33-3^, and reviewed by Anderson (38) and by 
Miller (39). These studies have shown that certain safety-modified nonreplicating retroviral 
vectors which contain the gene for the neomycin phosphotransferase enzyme (neo*^) can be 
used to mark and to trace cultured TIL-derived T-cells in vivo utilizing sensitive PCR detection 
methods. Neo^ marked TIL can be detected in both man and animals for a considerable time 
after in vivo administration. Therefore marking TIL with the neo*^ gene using retroviral 
vectors is an excellent way to study the homing and survival of these cells at the tumor site 
(33-37). Animal studies reported by Culver et al (33), reported that a positive signal for neo*^ 
was detected between 4 and 24 months In 3 of 4 monkeys who received IV injections of 
cultured T-cells infected with a neo^ containing retrovirus vector. Alexander et al (40) 
recovered TILs from mice that had been cured by treatment with TILs up to 1 19 days after 
they had received injections of these TILs. By administering rIL-2 concurrently to these 
animals TIL recovery was increased at all time points at the tumor sites. Moreover, when 
mice whose pulmonary metastases had been cured with TIL, were rechallenged with tumor, 
the donor TIL were found to concentrate at the tumor donor sites showing that these cells 
retained memory characteristics. Rosenberg reported a clinlca* trial involving patients with 
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Recombinant DNA Research, Volume 19 
