B. Pre-treatment evaluations: 
1. Complete medical history, complete physical examination and performance status 
assessment by a study physician. 
2. CBC, differential count, platelet count, PT, PTT. 
3. Chemistries: glucose, calcium, total protein, albumin, uric acid, phosphate, 
4. BUN, creatinine. 
5. Urinalysis. 
6. Pregnancy test (where indicated). 
7. Electrocardiogram, chest x-ray. 
8. p24 antigen level. 
9. CD4 counts. 
10. Quantitative PCR for viral load RNA analysis. 
11. Cryopreservation of 10 ml of serum and PBMC derived from 45 ml of 
peripheral blood. 
C. Gene Transfer Methods: 
In the gene transfer procedures outlined below, three measures will be employed to 
avoid activation of latent HIV present in patient peripheral blood lymphocytes. The non- 
nucleoside reverse transcriptase (RT) inhibitor nevirapine and CD4-pseudomonas 
exotoxin (CD4-PE40), will be included during the ex vivo expansion and transduction of 
patient lymphocytes. The latter agent is a fusion protein consisting of the HIV-envelope 
binding region of human CD4 linked to the translocation and ADP-ribosylation domains 
of Pseudomonas aeruginosa exotoxin A (50). Binding of one toxin domain allows entry of 
the lethal second domain into the cell; CD4-PE40 has been shown to selectively 
eliminate infectious HIV from cultures of human T cells lines when used in concert with 
reverse transcriptase inhibitors (51). Nevirapine belongs to the non-nucleoside RT 
inhibitors, a group of structurally diverse compounds that non-competitively inhibit 
HIV RT and viral replication at nanomolar concentrations, typically have therapeutic 
indices (assayed in cultured cells) of 1000 to 1, and are highly specific for HIV-1, 
lacking activity against HIV-2, SIV or murine reverse transcriptases (27,52,53). This 
synergistic combination of an agent selectively virustatic for HIV-1 and an agent 
selectively cidal for gpl 20-expressing cells will prevent amplification of HIV during 
the protocol without interfering with transduction by the PA31 7-packaged vector. 
Recombinant DNA Research, Volume 18 
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