DNA vial and mixed well by repeated inversion. The fmal concentration of the 
HLA-B7 plasmid DNA is 500 pg/ml. The amount injected into each tumor will be 
0.5 ml, for the highest dose of 250 pg. Lower doses, 10 and 50 pg, will be 
prepared in a similar fashion or formulated as dilutions with lactated Ringer's. 
Tumor lesions will be selected for treatment if they are accessible to 
intratumor administration by direct needle injection or intravascular 
catheter. These metastatic lesions will be located in the liver for colorectal 
adenocarcinoma. The study drug will be injected with the aid of 
sonographic visualization of the metastasis. 
Prior to injection following placement of the needle, gentle aspiration will 
be applied to the syringe to ensure that no material is injected 
intravenously. 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 48 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 
toxicides will be graded according to the WHO recommendations (see 
Appendix 2). 
16.0 Statistical Consideration and Methodology - Descriptive statistics only will be 
performed due to the small number of patients. 
17.0 Pathology Considerations 
17.1 Patients entering this trial will already have had histologically- 
documented metastatic colon carcinoma. Liver biopsy is carried out as 
part of this protocol including the initial one to demonstrate 
unequivocally by histologic examination of a frozen section that the 
metastasis selected for injection of the DNA/lipid complex is, in fact, 
metastatic cancer. Biopsy samples are never to be placed in fixative. Jill 
Piens, R.N. (507/284-4911, or pager 4-6362) to call Dr. J.S. Kovach's 
laboratory and state that a biopsy sample on this immunotherapy protocol 
has been obtained and is being held in a dry sterile container placed on 
ice to be picked up. 
17.2 For biopsies in which a core of tumor has been obtained, the sample will 
be subdivided and at least half frozen in liquid nitrogen. The remainder 
Recombinant DNA Research, Volume 18 
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