in BUN and creatinine, bilirubin and liver enzymes, hypertension and, after 
prolonged administration, pancytopenia. Minor side effects may include 
hirsutism, gingival hyperplasia and possible skin changes, mental depression, 
tremor, and hypertension. 
Serum samples will be obtained according the concurrently active protocols 
for CSP pharmacokinetic studies. 
3. Dose adjustment 
Rising BUN and creatinine and pancytopenia may require reductions of the 
dose of CSP. 
C. Generation and characterization of HIV-specific T cell clones for immunotherapy 
The methodologies employed to generate CD8 + HIV-specific T cell clones from HIV 
seropositive individuals pretransplant and to transduce the T cell clones with the HyTK 
retrovirus are detailed in Appendix 1. T cell clones transduced with HyTK and demonstrating 
HIV gag-specific cytolytic function will be characterized for 
1. Cell surface phenotype (CD3, CD4, CD8, CD 16) 
2. Glass I MHC restriction 
3. DL-2 dependence 
4. Correct structure of transferred HyTK vector DNA 
5. In vitro ganciclovir sensitivity 
6. Vaccinia or latent HIV infection by PCR for HIV gag DNA 
7. Absence of helper virus (S + L' and neovirus rescue assay) 
8. Mycoplasma, fungal and bacterial sterility 
The generation and in vitro growth of autologous HIV-specific T cell clones requires a single 
blood sample (60cc) from the HIV positive patient and the following from the HLA-identical 
HIV seronegative bone marrow donor: 
1 . 60 cc of peripheral blood obtained in the pretransplant work-up (required twelve 
weeks prior to the scheduled BMT). 
2. 3 mm punch skin biopsy obtained in the pretransplant work-up (required at least 
twelve weeks prior to scheduled BMT to establish a fibroblast line). 
3. Two leukaphereses to obtain and store PBMC for use as feeder cells. 
D. Administration of CD8 + HIV-specific T cell clones transduced with the HvTK retrovirus. 
1. Autologous T cell infusions will begin following engraftment as defined by an ANC 
of > 750/mm 3 providing Acyclovir has been discontinued for ^ 24 hours and the 
patient is not on ganciclovir. 
2. All T cells administered will be clonally-derived CD8 + class I MHC-restricted HIV- 
specific cytotoxic T cells generated from peripheral blood lymphocytes of the HIV- 
seropositive patient. 
3. Each patient will receive up to four intravenous infusions of HYTK-transduced T c at 
increasing cell doses given two weeks apart, according to the dose escalation scale 
outlined in Table 1 or patient tolerance. 
Recombinant DNA Research, Volume 15 
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