6.0 Nature of Procedures or Therapeutic Agents 
6.1 Study Design. 
This study is designed to determine whether the introduction 
of a transdominant form of Rev will prolong the survival of T 
lymphocytes in vivo which contain this gene. If resistance can 
be conferred upon these cells to HIV infection in vivo, then it 
will warrant further studies to optimize this molecular 
intervention for therapeutic purposes in AIDS patients. Briefly, 
the Rev M10 gene will be introduced into CD4 + cells after 
depletion of mononuclear cells and B cells using soybean 
agglutinin and removal of CD8 + cells with antibody-coated plates 
(see Appendix IV) . These CD4 + cells are -90% homogenous and will 
be stimulated in vitro. Two methods of stimulation will be used, 
each in a separate patient. In the first instance, cells will be 
stimulated with anti-CD3 and IL-2 as by standard methods 
(Appendix IV) . In the second case, cells will be stimulated with 
antibodies to CD3 and CD28 coated to tissue culture plates. 
These antibodies stimulate different signal transduction pathways 
to T cells, and have different growth stimulation properties. 
For example, CD3 and CD28 stimulation provide preferential 
proliferation of CD4 + cells while stimulation with CD3 and IL-2 
stimulates both CD4 and : CD8 populations simultaneously. 
In each case, a control gene will be introduced into a 
separate aliquot of cells which contain a frameshift which 
abrogates Rev M10 expression (ARev M10) . We will first analyze 
the effects of Rev M10 or ARev M10 on T cell survival after 
introduction into T cells by retroviral vectors. The structure 
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Recombinant DNA Research, Volume 18 
