Human endogenous retroviruses env gene expression and long terminal repeat methylation in colorectal cancer patients.

Human endogenous retroviruses (HERV) are remnants of exogenous retroviral infections, representing 8% of the human genome. Their regulation is based on the DNA methylation of promoters, the long terminal repeats (LTRs). Transcripts from HERV have been associated with cancers, but reports concerning HERV expression in colorectal cancer remain sporadic. Sixty-three patients with advanced stages of colorectal cancer were enrolled in this study. The expressions of HERV env gene, and HERV-H, -K, -R and -P LTRs and Alu, LINE-1 methylation levels, were investigated in the tumor, normal adjacent tissues, and, where possible, blood and plasmatic extracellular vesicles (EVs). Associations among HERV env expression, methylation status and clinical characteristics were evaluated. No differences were observed in HERV env gene expression levels among the clinical specimens, while Alu, LINE-1, HERV-H and -K LTRs were demethylated in the tumor compared to the normal adjacent tissues (p < 0.05).The HERV env gene was expressed in the EVs at of 54% (-H), 38% (-K), 31% (-R) patients. Association was not found between HERV env expression and LTR methylation, but significant higher expression of HERV-P and -R env was found in tumor tissues arising from the right colon. Our findings do not demonstrate significant overexpression of the studied HERV in colorectal cancer, but their association with tumor localization and specificity of the changes in DNA methylation of retroelements are shown. HERV sequences were packaged in the EVs and might be transferred from one cell to another.

Generation of tumor-specific cytotoxic T-lymphocytes from the peripheral blood of colorectal cancer patients for adoptive T-cell transfer.

This study designs a strategy for an adoptive cellular therapy (ACT) protocol based on the ex-vivo selection of autologous peripheral blood-derived CD8-enriched T-cells, stimulated with dendritic cells (DCs) that had been pulsed with apoptotic tumor cells to generate cytotoxic T lymphocytes (CTLs) with anti-tumor activity. Seventy-eight colorectal cancer (CRC) patients were enrolled in this study. Tumor tissues and peripheral blood (PB) were obtained at surgery. Tissues were mechanically dissociated and cultured to obtain a primary tumor cell line from each patient. DCs were derived from peripheral blood mononuclear cells (PBMCs) using magnetic positive selection of CD14+ monocytes. Anti-tumor CTLs were elicited in co-/micro-cultures using DCs as antigen-presenting cells, autologous apoptotic tumor cells as a source of antigens, and CD8+ T lymphocytes as effectors. Interferon-γ (IFN-γ) secretion was assessed by ELISpot assays to evaluate the activation of the CTLs against the autologous tumor cells. Primary tumor cell lines were obtained from 20 of 78 patients (25.6%). DCs were generated from 26 patients, and of them, corresponding tumor cell lines were derived from six patients. ELISpot results showed that significant IFN-γ secretion was detected after different numbers of stimulations for two patients, whereas weak secretion was observed for three patients. Despite difficulties due to contamination of several primary tumor cell lines with gut intestinal flora, the results suggest that the generation of tumor-specific CTLs is feasible from patients with CRC, and could be useful for supporting an ACT approach in CRC.

Italian multicenter experience of Roux-en-Y gastric bypass on vertical banded gastroplasty: four-year results of effective and safe innovative procedure enabling traditional endoscopic and radiographic study of bypassed stomach and biliary tract.

BACKGROUND: Cancer, perforation, and bleeding in the bypassed stomach after Roux-en-Y gastric bypass (RYGB) are rare, but serious, complications that need an early diagnosis. Our goal was to perform gastric bypass such that traditional endoscopic and radiographic study of the gastric remnant would be possible and, at the same time, obtain results in terms of weight loss equivalent to those found after standard RYGB. A previously published study demonstrated that complete occlusion of the gastrogastric outlet was not necessary to lose weight. We have developed an open RYGB-on-vertical banded gastroplasty procedure. METHODS: Since 2002, 289 patients with a mean age of 40.1 +/- 14.8 years, mean body mass index of 51.4 +/- 7.3 kg/m(2), and mean percentage of excess body weight of 107.3% +/- 36.7% underwent RYGB-on-vertical banded gastroplasty as their primary procedure. RESULTS: The follow-up examinations included radiographic and, if necessary, endoscopic studies at 6 and 12 months postoperatively and annually thereafter. Two cases of anastomotic ulcer were detected, one of which involved band erosion. The percentage of excess weight loss was 48.2% +/- 18.8% after 6 months and 59.0% +/-17.7%, 63.3% +/- 13.9%, 66.9% +/- 17.5%, and 70.0% +/- 17.7% after 1, 2, 3, and 4 years, respectively. The weight loss curve was similar to that for standard RYGB. CONCLUSION: The results of our study have shown that RYGB-on-vertical banded gastroplasty is as effective as traditional RYGB, while allowing for traditional radiography of the bypassed stomach in every patient. Endoscopy of the distal stomach and, therefore, the biliary tract, was also possible. These are the fundamental aspects of the procedure.