Finally, although we do not expect that the relatively small 
number of cells in this study will confer protective effects on 
immunologic function, we will nonetheless examine CD4 counts and 
viral load using a commercially available assay to quantitate 
serum viral RNA levels (Chiron) . 
11.2 Confirmation of Recombinant Gene Expression 
Several independent techniques will be used to evaluate the 
presence and expression of the recombinant gene in vivo. We have 
used several antibodies to Rev to detect the recombinant gene 
product in vivo by immunoprecipitation (16) . The presence of 
retroviral or plasmid DNA will be confirmed by PCR of DNA from 
peripheral blood lymphocytes or in selected lymph node specimen 
tissue, if available. RNA will also be isolated and examined for 
the presence of Rev M10 or ARev M10 mRNA by reverse transcription 
PCR or SI nuclease analysis. 
11.3 Analysis of Immune Response 
Cytolytic T cell activity will be evaluated by incubation of 
peripheral blood lymphocytes with 51 Cr labeled Rev M10 transduced 
EBV lines from the patient. The presence of antibody will be 
evaluated by FACS analysis of a matched pair of Rev M10 + or ARev 
M10 cell lines. In some instances, lymphocytes will be isolated, 
expanded in tissue culture, and analyzed for cytolytic function. 
11.4 Toxicity 
Toxic side effects of treatment will be analyzed and 
classified by the common toxicity criteria (Section 13) . 
Recombinant DNA Research, Volume 18 
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