be carried out by Dr. Gary Sudakoff, or one of his associates, prior to admission. 
Tumor lesions will be selected for treatment if they are accessible to Lntratumor 
administration by direct needle injection or intravascular catheter. These metastatic 
lesions will be located at any accessible site such as skin, lymph nodes, lung, liver, 
kidney or spleen for renal cell carcinoma. If necessary, the study drug will be 
injected with the aid of sonographic visualization of the metastasis. 
Once all pretherapy tests and procedures are completed, patients will be admitted to 
the Hematology/Oncology service at the University of Chicago prior to each 
intralesional injection. 
7.3 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 DMRIE 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 
DNA vial and mixed well by repeated inversion. The final concentration of the 
HLA-B7 plasmid DNA is 500 pg/ml. The amount of DNA injected into each tumor 
will range from lOpgm to 250 pg. Lower doses, 10 and 50 pg, will be prepared in a 
similar fashion or formulated as dilutions with lactated Ringer's. 
7.4 The appropriate volume of DNA/lipid mixture prepared in the University of 
Chicago Pharmacy will be injected into the pre-selected metastasis on day 1. The 
injection is done by the diagnostic radiologist using standard procedures in the 
Diagnostic Radiology Suite. The needle will be inserted under direct sonographic (or 
CT) visualization. A variable volume of lipid preparation (up to 4ml) will be used 
but all DNA concentrations will be kept constant per dose level. 
Prior to injection and following placement of the needle, gentle aspiration will be 
applied to the syringe to ensure that no material is injected intravenously. Up to 4ml 
of the DNA/lipid mixture will be injected as described above. Vital signs will be 
measured every 15 minutes prior to, during, and after the injection for at least two 
hours or until the patient is stable. If the systolic blood pressure drops below 80mm 
Hg, the injection will be terminated immediately, and the patient will be closely 
monitored until blood pressure is normalized. 
Immediately after the injection, a blood sample will be obtained to check serum 
enzymes, blood chemistries and cell counts, and to analyze by PCR for the presence 
of HLA-B7 Plasmid DNA in the peripheral blood. Every patient will be observed for 
23 hours and another blood collection will be drawn. If there are no complications, 
the patient will be discharged. If any abnormalities appear, the patient will be 
closely observed. All toxicities will be graded according to the WHO 
recommendations (see Appendix 2). 
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