1 . OBJECTIVES 
1 . 1 Primary 
1 . 1.1 
1.2 Secondary 
1 . 2.1 
1 . 2.2 
To assess the safety and tolerance of the adoptive transfer of anti-HIV 
cytotoxic, syngeneic peripheral blood T-lymphocytes in patients with HIV 
infection. 
To determine the activity of adoptively transferred, genetically marked 
syngeneic peripheral blood anti-HIV cytotoxic T-lymphocytes in immune 
function of patients with HIV infection. 
To determine longevity of the genetically marked CD8-I- lymphocytes in the 
peripheral blood of patients with HIV infection. 
2.1 Pre-clinical Studies 
2.1.1 In Vitro 
Numerous studies in rodents have demonstrated that the adoptive transfer 
of antigen specific CD8-f- T cell clones can eradicate viral infection such as 
influenza (33,34). In addition, anti-HIV specific human T cell clones have 
been successfully generated (Protocol 92-1-0035). However, since these 
clones recognize antigen in an MHC-restricted fashion, they are not 
applicable as therapeutic agents for use as adoptively transferred cells in the 
majority of humans. 
The adoptive immunotherapy-gene therapy approach utilized in this study 
redirects the antigenic-specificity of CD8-I- cytotoxic T cells to kill HIV 
infected target cells using non-MHC restricted "universal" T cell receptors 
(URs). Upon binding to the viral antigen, these anti-HIV specific URs 
initiate T cell activation, resulting in induction of effector functions, 
including cytolysis of the HIV-infected cell. In these chimeric receptors,' 
the cytoplasmic domain of zeta is fused to an extracellular domain of the 
targeting molecule, which confers specificity for HIV env protein. The UR 
derives its specificity from the extracellular domain of the human CD4 
receptor which recognizes the gpl20 moiety of HIV env, currently the 
only well characterized high affinity receptor for HIV. 
Expression of the CD4-UR in a model T cell line enabled the cells to initiate 
T cell effector functions such as cytokine secretion in response to HIV-env 
expressing target cells. These initial experiments served to demonstrate 
functionality of the anti-HIV URs in T cell activation. In the subsequent 
experiments, their ability to mediate T cell effector functions in primary 
human CD8+ T cells was evaluated. We therefore introduced the UR via 
retroviral transduction into human CD8-I- T cells purified by positive 
selection from the peripheral blood mononuclear cells (PBMCs) of normal 
donors. Purified populations of CD8-f- T cells were subsequently isolated 
that expressed surface UR as seen by fluorescence analysis, and have 
integrated provirus in the genome as demonstrated by Southern analysis. 
The HlV-specific UR was stably expressed at high levels on the surface of 
transduced CD8-I- T cells modified in this manner. The structure and 
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Recombinant DNA Research, Volume 19 
