Detection of micro-metastases by flow cytometry in lymph nodes from patients with penile cancer.

BACKGROUND: The tumor draining lymph node concept was first described in penile cancer for staging. Immunohistochemistry and histopathology evaluations are routinely used in clinical practice to examine lymph nodes for metastasis. However, these methods are time-consuming with low diagnostic accuracy and micro-metastases might be missed. In this study, we aim to evaluate detection of metastatic cells in draining lymph nodes by flow cytometry. METHODS: To assess the sensitivity of micro-metastasis detection by FACS (Fluorescence-activated cell sorting), HeLa cells were titrated into Peripheral blood mononuclear cells (PBMCs) and expression of pan-cytokeratin AE1/AE3 was analyzed. Single cell suspensions were separately prepared from 10 regional lymph nodes obtained from 5 patients with invasive penile cancer undergoing radical surgery and lymph node dissection. Lymph node dereived cells were examined for cell surface expression of EpCAM, E-cadherin and intracellular expression of pan-cytokeratin AE1/AE3 by FACS. RESULTS: Ten lymph nodes from 5 penile cancer patients were investigated in a head-to-head comparison between FACS and pathology examination of sections. All metastatic lymph nodes verified by pathology examination were also identified by FACS. Two additional lymph nodes with micro-metastases were diagnosed by FACS only. CONCLUSIONS: FACS analyses of pan-cytokeratin AE1/AE3 stained single cells from tumor draining lymph nodes can be used to detect micro-metastases in patients with penile cancer patients.

Immune responses against autologous tumor and human papilloma virus in lymph nodes from patients with penile cancer.

PURPOSE: Nearly half of penile cancers are related to human papillomavirus (HPV) infection. Investigations of tumor- and HPV-specific T cell reactivity in regional lymph nodes (LNs) from patients with penile cancer are warranted. MATERIALS AND METHODS: In this study, single-cell suspensions from LNs and peripheral blood from 11 patients with penile cancer were stained with antibodies for lymphocyte markers and analyzed by fluorescence-activated cell sorting (FACS). DNA was extracted from the tumor tissue and HPV status was investigated by PCR. RESULTS: T-cell reactivity against autologous tumor-extract and against the HPV-vaccine Gardasil® was tested by flow-cytometric assay of specific cell-mediated immune response in activated whole blood (FASCIA). CD4+/CD8+ ratios were significantly lower in HPV positive LNs (p<0.05). Immune responses to tumor extract assessed by blast transformation and expansion in vitro, of either CD4+ or CD8+ T-cells, were found in 9 of 13 LNs (69%). 5 of 6 tested patients demonstrated T cell recognition of tumor-associated antigen(s). In HPV-positive patients, dose-dependent T cell responses against L1 (late) HPV proteins (Gardasil vaccine) were demonstrated. CONCLUSIONS: LN-derived T cells from patients with penile cancer recognize tumor antigen(s) and in HPV-positive cases, there is a response against L1 (late) HPV proteins, being constituents of the Gardasil vaccine.

Experience of endoscopic transanal resections with a urologic resectoscope in 131 patients.

PURPOSE: Endoscopic transanal resection of rectal adenomas and other presumably benign lesions is not widespread. The purpose of this study was to evaluate the efficacy and the safety of endoscopic transanal resection. METHODS: Patients who underwent endoscopic transanal resection at three Stockholm hospitals between 1993 and 2004 were studied retrospectively with respect to patient and lesion characteristics, complications, follow-up time, and recurrence rates. RESULTS: One hundred eighty endoscopic transanal resection procedures were performed in 131 patients. The tissue diagnosis was adenoma in 160 operative cases, cancer in 12 operative cases, and hyperplasia, fibrosis, or normal mucosa in the remaining 8 operative cases. Among the patients with rectal adenomas, one endoscopic transanal resection was sufficient in 77 cases and in 16 cases the surgery was performed in more than one session because of the large size of the adenoma. In 27 cases there were recurrences that needed additional endoscopic transanal resection or other surgery. The median time until recurrence was seven months, but there were no recurrent rectal carcinomas. In 16 operative cases there were complications. Two patients had to undergo a Hartman's procedure as a result of a bowel perforation, and one patient had to be reoperated on because of bleeding. There were no perioperative deaths. The median follow-up time without recurrence was 32 (range, 0-67) months. CONCLUSIONS: Endoscopic transanal resection is a feasible and oncologically safe option for treatment of rectal adenomas, especially in cases where conventional transanal resection or transanal endoscopic microsurgery are unavailable or unsuitable because of the characteristics and localization of the lesion.