and lateral chest x-ray, CT scans and other indicated radiographic screening, Measurement of viral 
shedding, and immunological testing of pre-gene therapy blood for reactivity against adenovirus and 
the presence of cytotoxic T-lymphocytes. 
Pretransfection treatment 
Patients will be admitted to the Thoracic Surgery Service of the Hospital of the University of 
Pennsylvania (H.U.P.) and will undergo standard pre-operative evaluation and preparation. On Day 
1 , patients will undergo a diagnostic video thoracoscopy under either regional or general anesthesia. 
Utilizing standard thoracoscopic operative techniques and real-time imaging equipment, Dr. Kaiser 
and his assistants will visually inspect and photograph the pleural surfaces of the lung, 
mediastinum, chest wall, and diaphragm to document the extent and location (relative to standard 
anatomic landmarks) of tumor. Biopsies will be taken from parietal pleura at the apex, diaphragm, 
lateral chest wall, mediastinal pleura, and visceral pleura. Biopsy tissue will be evaluated both by 
frozen section, permanent histologic sections, and electron microscopy. Special stains using 
immunohistochemical techniques will be used to support the diagnosis of mesothelioma. Any pleural 
fluid removed at the time of this procedure will be evaluated by routine chemistry, microbiology and 
hematology, and cells will also be evaluated by cytopathology. A chest tube will be placed in the 
pleural space and placed to suction. The patients will be returned to the recovery room and their 
hospital room after surgery. 
Transfection 
The diagnosis of mesothelioma will be confirmed. On Day 2, one day post-videothoracoscopy, the 
patient will be moved to the GCRC, where he/she will remain for the remainder of the stay under 
strict isolation conditions. The appropriate dose of virus will be instilled into the chest tube 
dissolved in 50 ml of normal saline. We will begin by instilling the Ad. RSVt/r virus at a dose of 10^ 
plaque forming units (pfu) into 3 patients. If there is no toxicity observed, we will increase the 
dose by one log as described in detail below to a maximum dose of 1 0^ pfu. After 2 hours, the chest 
tube will be removed. Vital signs including blood pressure, pulse, temperature, and respiratory 
rate will be measured and recorded every 1 5 min for the first hour, every 30 minutes for the next 
two hours, every one hour for the next six hours, and every four hours for the rest of the week 
post-transfection. 
Post-transfection Protocol 
Patients will be observed closely on Day 3 and Day 4. On Day 5, the patient will be taken 
to the operating room to have a repeat videothoracoscopy performed under local anesthesia. At this 
time biopsies (3-5) will be taken at standardized locations in order to determine the efficacy and 
characteristics of gene transfer (See Specific Aim #3). Additional biopsies will be taken where the 
pleura appears “normal". The location and orientation of each biopsy specimen will be marked by 
Drs. Kaiser and Litzky. Biopsy specimens will then be evaluated by routine staining, 
immunostaining, in situ hybridization, and PCR analysis. The patient will be returned to the CRC 
after surgery and remain in isolation. 
On Day 6, the patient will begin receiving ganciclovir at a dose of 5 mg/kg via IV infusion 
over one hour given BID. This will continue for 1 4 days with monitoring as described below. On 
Day 20, after the last dose of GCV, the patient will be discharged and followed closely as an 
outpatient via the schedule below. Further therapy will be at the discretion of the treating physician 
and be determined by the clinical situation. For example, the patient may require pleural sclerosis 
for the control of symptoms associated with an effusion. 
While in the hospital the patient will have his/her sputum, nasal swab, urine and stool 
analyzed for replication competent adenoviruses of any serotype using standard adenovirus assays. 
These samples will also be evaluated for HSVt/r recombinant virus using a F>CR assay. In the unlikely 
event that the patient continues to shed recombinant adenovirus in the airway, he/she will be kept in 
the hospital for a longer period of time. 
The following monitoring schedule will be used: 
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