Patients meeting the criteria for partial response at 4 weeks after their single 
injection on Schedule A or after their last injection on Schedule B may be 
retreated once. 
14.0 Pharmacologic/Immunologic Studies 
14.1 Immunochemical staining will be done in the Immunohistochemical Core 
Laboratory of the Mayo Cancer Center under the direction of Dr. Patrick 
Roche. Pre- and post-treatment tumor cells are stained with anti-HLA-B7 
antibodies, ME-1, BB7.1, and GSP5.3 (G. Nabel, personal communication) 
to look for expression. 
14.2 Presence of DNA from the HLA-B7 gene will be assessed by PCR 
amplification of cells obtained by biopsy of the treated site on days 8, 15, 
and 29 after injection of DNA/lipid complex. Genomic DNA is isolated 
by standard methods and a portion of the HLA-B7 gene is amplified and 
sequenced. Several primer sequences may be used (G. Nabel, personal 
communication). 
14.3 Development of circulating antibodies to HLA-B7 will be evaluated. 
Autologous peripheral blood B lymphocytes will be EBV immortalized 
and subjected to in vitro gene transfer with the DNA/lipid complex, these 
autologous cells expressing the HLA-B7 gene will be used to assess the 
specificity of antibody response to the in vivo transfer of the gene. 
Evidence of cytolytic T-cells will be assessed if a sufficient amount of 
material is available for successful expansion of infiltrating T-cell 
population from cells in biopsy of the metastasis. These studies will be 
carried out in the Cellular Immunology Laboratory under the direction of 
Dr. Homburger. 
15.0 Drug Information 
The study drug (VCL-1005) will be supplied by Vical as two sterile vials 
containing (i) HLA-B7 plasmid DNA, and (ii) DMRIE/DOPE lipid mixture. 
Diluent will be lactated Ringer's. All components will be stable for at least eight 
weeks under recommended storage conditions (DNA -20°C, DMRIE/DOPE 4°C). 
The materials will be supplied by Vical Inc. 
This study drug is composed of plasmid DNA coding for the complete human 
MHC HLA-B7 formulated with the cationic lipid mixture DMRIE/DOPE (lipid 
complex formulation). The DNA concentration is 1.0 mg/ml (see Investigator's 
brochure for complete details of product characteristics and preparation). 
Lactated Ringer's is readily available at the site. 
DNA/lipid complexes are prepared immediately prior to administration. DNA is 
supplied in 1.0 mg/ml concentration in 400 pi lactated Ringer's solution. Lipid 
(DMRIE/DOPE) is supplied as a dried film. Each vial contains 77 pg DMRJE and 
90 pg DOPE. Each vial is reconstituted with 400 pi lactated Ringer’s solutions by 
vortexing until homogeneous. The contents of the lipid vial is transferred into the 
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Recombinant DNA Research, Volume 18 
