indication of an autologous activation of the immune system. As a control, the same buffy layer will be 
incubated with the non-transfected parental glial cells from the primary tumor in the presence of 
tritiated thymidine. A significant difference in tritium incorporation between the two cultures will 
indicate activation of the immune system. As a positive control nontransfected glioblastoma cells will 
be incubated with buffy layer from a different individual and tritiated thymidine incorporation 
measured. We expect in this system a significant incorporation of thymidine into DNA in the cells of 
the buffy layer, since this is equivalent to an allogenic reaction. This information will be correlated 
with the dose of cells injected, and used to predict the patient's response. 
- Data analysis 
The initial toxicity trial is descriptive in nature. The magnitude of initial response in both 
clinical and laboratory analyses will enable us to determine the power needed for adequate sample 
sizes in a larger, randomized clinical trial to follow. In an expanded study, decision whether outcome is 
a success or failure would be based on measurement of tumor volume by MRI and on longevity. Factors to 
be paired in outcome analysis will be age, sex, tumor type and stage, and the Kamofsky performance 
status. The data for this study will be obtained and coordinated by the investigators and a nurse 
assistant dedicated to this project. Records will be managed through the facilities of the Ireland 
Cancer Center. 
2. Patient Selection (Population) 
Size and Source of Study Population and Mechanism for Contacting Subjects 
Patients older than 18 years of age will be recruited from the neurosurgery services of University 
Hospitals of Cleveland, Cleveland Veteran's Administration Hospitals and related hospitals of the 
University Hospitals ' group. Patients with newly diagnosed glioblastoma multiforme will be 
considered for experiemental treatment. Our past experience with the prevalence of glioma at these 
institutions suggests we would be able to recruit 50 patients per year with glioblastoma who meet the 
entrance criteria defined for this study. 
Number to be studied and design of experiments. Twelve patients will be studied. 
The patients will be divided into three experimental groups consisting of four patients each. There 
will be four patients at each cell dosage level. This study is designed to elicit evidence of toxicity. 
Direct evidence of efficacy will be sought in a subsequent randomized clinical trial. 
Patient Dose of transfected. 
No. irradiated cells 
4 10 X 10 6 
4 50 X 10 6 
4 200 X 10 6 
Treatment Schedule 
Day 1 
2s 
Wk 4 Wk 12 
The first cohort of four patients with tissue diagnosed glioblastoma multiforme will be treated 
according to the above schedule with 10 x 10^ cells obtained in cell culture from self and as modified by 
transfection with the pAnti-IGF-I vector. Prior to subcutaneous injection, the cells will be irradiated 
with 5,000 cGy delivered by cobalt source. In our experience this treatment is sufficient to bring about 
loss of the capacity to proliferate or to incorporate ^H thymidine into DNA in cultured rat or human 
cells. When non-transfected rat glioma cells are irradiated with this dose they also lose the ability to 
product subcutaneous tumors in syngeneic animals. 
If unacceptable toxicity is not obtained in the first cohort of patients treated, the second and third 
cohorts of of 4 patients each would receive the increased doses of cells in sequential order. Each cohort 
of patients would be separated in time by an interval of three months in order to determine toxicity 
and side effects before proceeding to the greater dose level. 
Recombinant DNA Research, Volume 18 
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