6.2 Confirmation of Gene Transfer and Expression 
Needle biopsy of the injected nodule will be performed after 
administration of local anesthesia prior to injection and 
subsequently to treatment (preferably at day 3). A portion of 
this tissue will be processed to obtain DNA for PCR analysis. 
The remaining tissue will be processed for pathologic analysis 
and immunohistochemical and/or immunof luorescent staining. If 
sufficient material can be obtained, RNA PCR analysis will also 
be performed. For internal organs, CT or ultrasound guided thin 
needle biopsies will be obtained when possible. 
6.3 Analysis of Immune Response 
Evidence of gene transfer can also be obtained indirectly by 
examination of the specific immune response to HLA-B7. The 
analysis will be performed as follows: two weeks prior to the 
initial treatment, a blood sample will be obtained to derive 
lymphocytes which will be immortalized using the Epstein-Barr 
virus. An aliquot of these cells will be further infected with 
an amphotropic HLA-B7 retroviral vector, and expression will be 
confirmed on the cell surface. These cells will subsequently be 
used in the laboratory as target cells for the cytolytic T cell 
assay. At no time will these cells be brought into the same 
building where the patient is being treated. These cells and 
laboratory experiments will be performed in MSRB II, Room 3560, 
whereas the patient's clinical treatment will be taking place in 
the Clinical Research Center of The University of Michigan 
Medical Center Hospital. 
Recombinant DNA Research, Volume 17 
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