Cytolytic function of transduced and ex vivo expanded T-cells will be determined by 
microlymphocytotoxicity assays employing 293 cells (human embryonic kidney line) 
expressing gpl20 as cytolytic targets. Tests for sterility, phenotype, function, and 
percent of transduced cells will be performed by BRI. 
5.5 Preparation of Cells for Infusion 
Approximately 2-3 weeks prior to the expected day of cell infusion, aliquots of 
stocked, cryopreserved cell cultures will be thawed and grown in a secondary 
expansion step to the required dose in accordance with treatment assignment (aiming 
for >1 X 10^® cells) in humidified incubators under 5% CO 2 at 37°C. Forty-eight hours 
prior to each scheduled harvest of the expanded cell cultures for subsequent infusion, a 
representative number of expansion bags will be tested for sterility as before. Once cell 
cultures attain the required number, cells will be aseptically harvested and pooled for 
infusion using a specialized cell harvester (Baxter or Dupont) and a Baxter Life-Cell™ 
Tissue Culture gas-permeable bag of a 1 liter capacity. Cells are then filtered through a 
170 micron filter in a solution of physiologic saline containing 2% human albumin, in 
preparation for infusion into the patient. Standard tests for sterility and percentage of 
transduced cells will be performed. Infusion will proceed if the proportion of 
CD8-f-CD4-i- phenotype exceeds 70% of the expanded cells. 
Units of expanded cells ready for infusion will be transported to the NIH Clinical 
Center by the BRI courier, under the conditions outlined in Appendix B. 
TREATMENT PROCEDURES 
6.1 Treatment Period I 
Treatment groups are shown in the table below. For cohort 0, lymphocytes pheresed 
from the seronegative twins will be fractionated into CD 8 - 1 - cells and expanded ex vivo, 
but without genetic modification. The corresponding seropositive twins will receive a 
single infusion at a cell dose of 1 x 10*®. For cohorts 1-3, the seronegative twin will 
likewise undergo apheresis, and the lymphocytes will be fractionated into CD 8 -f- cells, 
activated, transduced with a retrovirus carrying the gene for the chimeric protein 
targeting HIV-infected cells, and expanded ex vivo. The initial 6 corresponding 
seropositive twins will receive a lymphocyte infusion of 1 x 10^ cells (cohort 1). If at 
the first dose level no immediate dose-limiting toxicity has manifested in the minimum 
numbers of patients outlined in Section 1 1.1.3, randomization will proceed to cohort 2 
(1 X 10^ cells). The same rules of advancement will be followed for cohort 3 (1 x 
10*®). Following the single infusion of cells, patients will be observed through 8 
weeks for safety. 
Cohort 
Lymphocyte Dose 
Planned 
Number of 
IV infusions 
Minimum 
Number of 
Patients 
0 
1 X 10*® unmodified T-cells 
1 
6 
1 
1 X 108 
UR T-cells 
1 
6 
2 
1 X 109 
UR T-cells 
1 
6 
3 
1 X 10*® 
UR T-cells 
1 
6 
HIV-infected patients will be admitted to the Warren G. Magnuson Clinical Center 
inpatient wards for the first single infusion in Period I. An intravenous catheter will be 
Recombinant DNA Research, Volume 19 
