E-cadherin expression in bladder cancer using formalin-fixed,paraffin-embedded tissues: Correlation with histopathological grade,tumour stage and survival

To determine the potential prognostic value of epithelial cadherin (E-cadherin), a Ca²⁺-dependent cell-cell adhesion molecule, we have analysed its immunoreactivity and cellular localisation in 67 transitional cell carcinomas (TCC) using an avidin-biotin immunoperoxidase technique on formalin-fixed, paraffin-embedded tissues. These results were correlated with histopathological grade, tumour stage, presence of metastases and survival. In addition, 10 cystitis and 11 normal bladder biopsies were evaluated as controls. E-cadherin was expressed in a normal membranous pattern in all normal and 7 of 10 cystitis biopsies. Loss of normal surface E-cadherin was expression was found in 3 of 15 superficial tumours and in 48 of 52 invasive cancers. Abnormal immunoreactivity was strictly related to tumour differentiation and stage. Fifteen of 20 well-differentiated (grade 1) tumours showed preserved membranous E-cadherin immunoreactivity, while 46 of 47 moderate and poorly differentiated tumours (grades II and III) demonstrated abnormal staining patterns. Loss of membranous E-cadherin immuno-reactivity was also associated with advanced tumour stage. There was a significantly higher 5-year survival rate for patients with preserved membranous staining compared with patients with abnormal staining. © 1995 Wiley-Liss, Inc.     

Delayed disease progression after allogeneic cell vaccination in hormone-resistant prostate cancer and correlation with immunologic variables.

PURPOSE: There are a significant number of patients with asymptomatic hormone-resistant prostate cancer who have increasing prostate-specific antigen (PSA) levels but little or no evaluable disease. The immunogenicity and minimal toxicity associated with cell-based vaccine therapy makes this approach attractive for these patients. EXPERIMENTAL DESIGN: We have evaluated a vaccine comprising monthly intradermal injection of three irradiated allogeneic prostate cell lines (8 x 10(6) cells each) over 1 year. The first two doses were supplemented with bacille Calmette-Guérin as vaccine adjuvant. Twenty-eight hormone-resistant prostate cancer patients were enrolled. Patients were assessed clinically and PSA levels were measured monthly. Radiologic scans (X-ray, computed tomography, and bone scan) were taken at baseline and at intervals throughout the treatment period. Comprehensive monthly immunologic monitoring was undertaken including proliferation studies, activation markers, cytokine protein expression, and gene copy number. This longitudinal data was analyzed through predictive modeling using artificial neural network feed-forward/back-propagation algorithms with multilayer perceptron architecture.RESULTS: Eleven of the 26 patients showed statistically significant, prolonged decreases in their PSA velocity (PSAV). None experienced any significant toxicity. Median time to disease progression was 58 weeks, compared with recent studies of other agents and historical control values of around 28 weeks. PSAV-responding patients showed a titratable T(H)1 cytokine release profile in response to restimulation with a vaccine lysate, while nonresponders showed a mixed T(H)1 and T(H)2 response. Furthermore, immunologic profile correlated with PSAV response by artificial neural network analysis. We found predictive power not only in expression of cytokines after maximal stimulation with phorbol 12-myristate 13-acetate, but also the method of analysis (qPCR measurement of IFN-gamma > qPCR measurement tumor necrosis factor-alpha > protein expression of IFN-gamma > protein expression of interleukin 2). CONCLUSIONS: Whole cell allogeneic vaccination in hormone-resistant prostate cancer is nontoxic and improves the natural history of the disease. Longitudinal changes in immunologic function in vaccinated patients may be better interpreted through predictive modeling using tools such as the artificial neural network rather than periodic "snapshot" readouts.     

Surgical management of primary retroperitoneal hydatid cyst

Hydatid disease is caused by the cystic stage of infestation by Echinococcus granulosus. Most hydatid cysts occur in the liver followed in frequency by the lung but no site is immune. Hydatid disease of the retroperitoneum is a rare condition. A case study is described here of a 30-year-old female with diagnosis of hydatid cyst of retroperitoneum which was treated surgically.     

Cryopreserved dendritic cells for intratumoral immunotherapy do not require re-culture prior to human vaccination

Dendritic cell (DC) immunotherapy for cancer has shown great promise so far. The ability to deliver dendritic cells directly into tumours where they are capable of acquiring tumour antigens prior to stimulating specific T cell responses has been demonstrated both in animal models and human patients. Clinical grade DCs can be grown from peripheral blood monocytes in the absence of foetal calf serum (FCS) and cryopreserved to generate plentiful identical aliquots thus avoiding repeated venesection. However, the approach is still limited by the necessity to return thawed DCs to culture prior to injection. It would be more advantageous to directly inject the DCs whilst still in the freezing medium and thus prevent the need for further manipulation. Whilst several reports have shown that cryopreserved DCs can survive for over 72 h when returned to culture, there is no information regarding the longevity of cells maintained in the freezing medium after thawing. In this report we have shown that DCs may remain in freezing medium for up to 1 h without affecting their survival, phenotype or function. This period of time is sufficient to allow for any delays incurred between the preparation of the DCs and time taken to be administered within a standard clinical setting. This study demonstrates that clinical grade DCs can be cryopreserved and thawed whilst retaining the ability to acquire exogenous antigenic material required for intratumoural immunotherapy. The survival of these cells within the freezing medium without the requirement for re-culture expands their availability for administration directly to the tumours of patients in non-specialist centres that do not have the appropriate facilities for DC re-culture.     

Successful live cell harvest from bisected sentinel lymph nodes research report

Sentinel lymph nodes provide an excellent opportunity to study early immune responses to cancer. However, harvesting live cells has not previously been possible, because it conflicts with the need to preserve tissue for histological interpretation. This study used scrape cytology on 26 sentinel and 8 non-sentinel nodes, harvested from 17 stage I/II melanoma patients undergoing sentinel node biopsy. Numbers of viable cells harvested before and after cryopreservation were measured and the effect on subsequent histology assessed. The mean number of cells harvested from 26 sentinel nodes was 7.06 x 10(6) (range 0.1-32.2), with a mean viability of 99.5% (range 87-100, lower 95% CI 98.5%). Furthermore, counts and viabilities were well maintained after cryopreservation. Flow cytometry confirmed CD3+, CD20+ and lineage-1-/HLA-DR+ subpopulations, consistent with T-lymphocytes, B-lymphocytes and dendritic cells, respectively. Importantly, there was no discernible change in histological detail and the proportion of positive sentinel nodes remained unchanged. This technique will allow more functional and quantitative approaches to sentinel lymph node research.     

Adenoviral strategies for the gene therapy of cancer

Despite slow clinical progress, efforts to develop specific nontoxic cancer gene therapies are increasing exponentially. Adenoviral vectors are one of the most popular vehicles for gene transfer currently being used in worldwide clinical trials for cancer. Over the past decade our knowledge of the adenoviral life cycle together with the discovery of novel tumor antigens has permitted the targeting of adenoviral vectors to specific tumors. Targeting adenoviral vectors to tumors is crucial for their use in clinical applications in order to allow for systemic administration and the use of reduced vector doses. In addition, novel approaches to tumor killing have also been explored, which will have greater potency and selectivity than currently available treatments such as chemotherapy or radiation. This review discusses the basic concepts behind the use of adenoviral vectors for cancer gene therapy and their potential for clinical application, as well as ongoing and completed clinical trials.     

Gene therapy for head and neck cancer

The prognosis of patients with advanced head and neck cancer has not changed significantly in the last twenty years, despite concerted efforts to optimize treatment using conventional modalities such as surgery, radiotherapy and chemotherapy. Novel therapeutic approaches based on our increasing understanding of the molecular changes that underlie the development of cancer have the potential to alter this situation. Gene therapy involves the delivery of genetic sequences in to tumour or normal cells for a therapeutic purpose. A number of viral and non-viral vectors have been developed that have the ability to deliver therapeutic genes specifically to tumours. These therapeutic genes can exert their effects by correcting existing genetic abnormalities, by killing cells directly or indirectly through recruitment of the immune system. In this review, the various gene therapy strategies that are under development are presented with particular reference to the treatment of head and neck cancer.