Protocol ID93-008 
Page 15 
after injection, will be 5x10®/ 10^2 = 5x10-4. Our detection methods employing PCR have a 
sensitivity of lx 10'® cells so this procedure would be capable of detecting any marked cells 
that are among the sample sites under the null hypothesis that the CD8+ TILs are 
homogeneously distributed after injection. Assuming a 0. 1 gram tissue sample contains a 
approximately 10® CD8+ TIL (1% CD8+ T cells), the expected number of marked cells is at 
least 50, since we have argued that the proportion of marked lymphocytes in a patient is 
5x10-4 if they were homogeneously distributed. It is reasonable to assume that the number 
of cells marked in each tissue sample follows a Poisson distribution with a mean of 50. This 
means that the probability of observing at least one marked cell exceeds .9999 and if no 
marked cells are detected, the 95% confidence interval for portion of cells marked at that 
site is (0 to 3x10'®). Hence, the probability of a false negative is extremely low for each 
patient. 
In the event that there is specific uptake neo^^ marked TIL will not be equally distributed 
and substantial numbers will home into the tissue, much higher than those anticipated in 
normal tissues (postulated at 50 neo^^ marked TIL/0.1 gm). Therefore with the PCR 
detection technique that we have established, it would be easy to identify and quantitate 
neo*^ marked TIL that home to the tumor. PCR quantification in tumor and normal control 
sites will be performed according to the anal3ftic methods of Morgan et al. (43). A successful 
outcome in this experiment is defined when the mean number of marked cells in tumor 
tissue is significantly larger than the mean number of marked cells in either the blood and 
the normal tissue. The mean referred to here is the average of the measurements taken in 
triplicate. If the first five patients exhibit a successful response, then there is significant 
evidence at the .05 level to reject the null hypothesis that the response rate is less than or 
equal to 1/2. In other words, if the first five patients exhibit a successful response, then 
there is evidence at the .05 level of significance that the success rate is greater than 1/2, 
and this part of the clinical trial can be terminated. In order to collect evidence that the 
probability of success is less than .30. 9 evaluable patients are needed. If all 9 patients 
have an unsuccessful outcome: i.e., the mean number of marked cells in tumor tissue is 
less than or equal to the mean number of marked cells in both the blood and the normal 
tissue, then there is evidence to reject the null hypothesis that the probability of success is 
greater than or equal to .30. That is, if all 9 patients have an unsuccessful outcome, then at 
the .05 level there is evidence that the probability of success is less than .30. 
1 .0 POTENTIAL SIDE EFFECTS AND REPORTING OF ADVERSE REACTIONS 
11.1 Reporting of Adverse Reactions 
Adverse drug reactions (ADRs) to GlNa neo^^ are to be reported promptly to the 
Investigational Drug Branch. ADR reports are required even if there is only a suspicion 
of a drug effect. If in doubt call the Investigational Drug Branch of the NCI at 301-230- 
2330. 
11.1.1 Adverse reactions are to be reported to the NCI in writing using the “NCI 
Adverse Reactions Form for Investigational Agents" appended to ID92-015 
(T92-0093) within 10 working days. 
11.1.2 Grade 4 reactions and patient deaths while on treatment are to be reported to 
NCI by phone within 24 hours. A written report is to follow within 10 working 
days. 
Recombinant DNA Research, Volume 19 
[473] 
