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Purpose: Vaccinia virus is a DNA poxvirus previously used as a vaccine to eradicate smallpox. The virus has a high efficiency of infection, replicates in the cytoplasm without chromosomal integration and can transport a large amount of recombinant DNA without losing infectivity. Therefore, it is an excellent choice as a vector for gene delivery in vivo. Large quantities of vaccinia have been injected into dermal, subcutaneous and peripheral lymph node melanoma metastases without significant side effects, and with efficient infection of the tumor cells and recombinant gene transfection. To determine if vaccinia, when given intravesically, can effectively infect bladder mucosa and tumor with acceptable toxicity, we performed a phase I trial of intravesical vaccinia in patients with muscle invasive transitional cell carcinoma before radical cystectomy., Materials and Methods: After documenting immune competence and demonstration of a major reaction after revaccination, patients received 3 increasing doses of intravesical Dryvax vaccinia virus (Wyeth-Ayerst Laboratories, Philadelphia, Pennsylvania) that was provided by the Centers for Disease Control. Approximately 24 hours after the third dose, cystectomy was performed and the tissue was examined microscopically., Results: There were 4 patients who were treated. The 3 patients who received the highest doses (100 x 106 plaque forming units) had significant mucosal and submucosal inflammatory infiltration by lymphocytes, eosinophils, and plasma cells into tumor and normal tissue. Dendritic cells were recruited to the site after exposure to the vaccinia. Significant mucosal edema and vascular ectasia were seen. Tumor and normal urothelial cells showed evidence of viral infection, including enlarged vacuolated cells with cytoplasmic inclusions. There were no clinical or laboratory manifestations of vaccinia related toxicity except mild dysuria. Of the 4 patients 3 survived and were free of disease at 4-year followup., Conclusions: Our study demonstrates that vaccinia virus can be administered safely into the bladder with recruitment of lymphocytes and induction of a brisk local inflammatory response. To our knowledge, this is the first report of direct delivery of live virus into the human bladder. The role of wild type vaccinia as immunotherapy for bladder cancer warrants further study. Furthermore, these data support the exploration of recombinant vaccinia as a putative gene therapy vector for intravesical infection and transfection of bladder tumor cells with cytokine or other genes, an approach that our group pioneered and most recently studied in patients with superficial melanoma.

The mucin lining of the bladder is thought to serve as a primary defense mechanism against bacterial colonization, and has recently been implicated in the urothelial resistance to carcinogenic insult. We have isolated a unique glycoprotein fraction (GP1) of this lining from the normal rabbit bladder which may have a function in preventing bacterial adherence and colonization in the urinary tract. This glycoprotein has been shown to bind to a wide range of uropathic bacteria. The present study examines changes in the bladder's antibacterial defense mechanisms as measured by GP1 expression in the neoplastic state. Using an anti-GP1 serum, immunohistochemical staining was performed on 20 paraffinized and fresh frozen transitional cell carcinomas ranging from low grade, superficial tumors to high grade, invasive tumors. The presence of GP1 was seen throughout the mucosal layer in normal specimens with increased amounts noted towards the mucosal surface. Progressively decreased expression was noted with increasing grades of all transitional carcinoma specimens. Mucosal field changes in GP1 expression were not noted in any of the patients. Intestinal mucosal controls failed to detect the presence of GP1. These studies suggest that the expression of GP1 decreases with tumor dedifferentiation and that bladder tumorogenesis may serve a role in handicapping the bladder's primary antibacterial defense mechanism.

From 1978 through 1987, 78 patients with carcinoma of the bladder were treated with combined pre- and postoperative adjuvant radiation therapy. All were given a single dose of pre-operative radiation therapy, 500 cGy, either on the day of or the day before cystectomy. Histological staging on the cystectomy specimens according to the TNM classification system was performed. Forty patients with Stage P2 (high grade III and IV), P3A, P3B, P4A, or N+ underwent planned high dose postoperative radiation therapy (4000-4500 cGy) in 5 weeks. The whole pelvis was treated with conventional fractionation of 180 cGy 5 days per week. Median follow-up was 52 months, with 36 months minimum follow-up. There was a 67% overall 5-year survival, and those with P1 and P2 (Grade I and II) had an 84% 5-year survival. Survival for patients with P2 tumor (Grade III and IV), P3A, P3B, and P4/N+ stages was 57%, 56%, 39%, and 50%, respectively. Bowel obstructions developed in 8% of patients who received no postoperative radiation therapy and 37% in those who did. Genitourinary complication rates were similar in both groups, 13% in the group that received no postoperative radiation therapy and 10% in the group that did. Although the planned approach of combined pre- and postoperative radiation therapy for unfavorable stages of bladder cancer is associated with a better than 50% 5-year survival rate (except in Stage P3B cancer), the bowel toxicity was unacceptably high.

Thirty-nine patients with invasive bladder cancer were treated using a new approach to adjuvant radiation therapy. All patients had histologic evidence of bladder muscle invasion on biopsy and were considered suitable for surgical resection. Low-dose preoperative radiation (500 rad) was delivered either on the day of or the day before cystectomy. In most instances, a radical cystectomy was performed and patients were then stratified according to pathologic stagings. Those patients with pathologically good prognostic indicators, Stage B1 low grade (I or II) disease, were followed up with no further treatment. Patients at high risk for local recurrence and pelvic lymph node involvement, Stages B1 high grade (III or IV), B2, and C, were given aggressive postoperative radiation (4,500 rad in five weeks). Patients with advanced disease, Stage D2, were given palliative therapy. All patients received the dose of preoperative radiation as planned. Preliminary results indicate that combined pre- and postoperative radiation in bladder cancer gives excellent local control of disease. A significant potential for improved long-term survival of patients has been observed.

Peanut lectin (PNA) binds to D Gal B (1-3) D Gal NAc which is the purported antigenic determinant of the so-called T blood group antigen (TAg). We used PNA in an indirect immunofluorescent technique to study TAg in transitional cell carcinomas of the bladder (TCC). All 14 control tissues failed to express TAg, whereas all controls (100%) expressed TAg on the cell membrane after prior treatment with neuraminidase. Sixty-nine percent of Stage O TCC expressed TAg, whereas 100% of cases expressed TAg after treatment with neuraminidase. Staining patterns noted were: granular intracytoplasmic, intracytoplasmic globular, and membrane. There was no correlation between TAg positivity or negativity and subsequent muscle wall invasion in the Stage O TCC group. Fifty percent of Stage B or greater TCC expressed TAg, whereas 27 of 30 (90%) cases expressed TAg after neuraminidase treatment. TAg-staining patterns noted in Stage B tumors were: membrane, granular intracytoplasmic, intracytoplasmic globular, and glycocalyx. Only 47% of Grade III TCC expressed TAg compared with 72% of low-grade TCC. The authors conclude that the expression of TAg reflects the neoplastic transformation of bladder epithelium. The data also suggest that in TCC, TAg positivity is most likely due to incomplete synthesis of MN blood group glycoprotein, as evidenced by loss of a terminal sialic acid, and that there is also evidence of cytostructural "relocalization" of TAg.

Forty patients with stage O papillary transitional cell carcinoma of the bladder were tested for the presence or absence of ABH blood group substances. Of 25 tumors with antigens 21 (84 per cent) remained stage O, with a mean followup of 13.3 years. Of 15 tumors devoid of blood group substances 11 (73.3 per cent) progressed to stage B or greater in a mean of 4.3 years. Patients without antigens whose tumor remained stage O were followed for a mean of 10 years. Two patients with initially positive tumors subsequently showed loss of antigens and invasion. In 7 of 8 patients the benign epithelium showed a greater concentration of blood group substances than did the corresponding tumor. The presence or absence of blood group substances can be used to predict the biological behavior of stage O transitional cell carcinoma.

Ninety-two patients with bladder cancer have been treated with combined pre- and postoperative radiation in a Radiation Oncology Study Group (RTOG) Phase I-II study and at Thomas Jefferson University Hospital. Patients with invasive bladder cancer were entered into the study and given low-dose preoperative radiation (500 rad) to the whole pelvis, either on the day of or the day before cystectomy. Following surgery, patients were pathologically staged. Patients with stage B1 (T2) (grade 3 or 4), stage B2 and C (T3) tumors were given 4500 rad in 5 weeks postoperative radiation. Follow-up in the study ranges from a minimum of 24 months to 5 years, with a median of 36 months. Incidence of complications was 15% (14/92). The 4-year actuarial survival (Kaplan-Meier) by stage of disease is 68% for stage B1 (T2) (grade 3 or 4), 78% for stage B2, and 57% for stage C. These survival results appear to be better than those obtained with other approaches of adjuvant therapy and/or surgery in comparable histopathologically staged patients.