Tumor lysate-loaded Bacterial Ghosts as a tool for optimized production of therapeutic dendritic cell-based cancer vaccines.

Cancer immunotherapy with dendritic cell (DC)-based vaccines has been used to treat various malignancies for more than two decades, however generally showed a limited clinical success. Among various factors responsible for their modest clinical activity is the lack of universally applied, standardized protocols for the generation of clinical-grade DC vaccines, capable of inducing effective anti-tumor immune responses. We investigated Bacterial Ghosts (BGs) - empty envelopes of Gram-negative bacteria - as a tool for optimized production of DC vaccines. BGs possess various intact cell surface structures, exhibiting strong adjuvant properties required for the induction of DC maturation, whereas their empty internal space can be easily filled with a source tumor antigens, e.g. tumor lysate. Hence BGs emerge as an excellent platform for both the induction of immunogenic DC maturation and loading with tumor antigens in a single-step procedure. We compared the phenotype, cytokine secretion profile, functional activity and ability to induce immunogenic T-cell responses in vitro of human monocyte-derived DCs generated using BG platform and DCs matured with widely used lipopolysaccharide (LPS) plus interferon-γ cocktail and loaded with tumor lysate. Both approaches induced DC maturation, however BG-based protocol was superior to LPS-based protocol in terms of the ability to induce DCs with a lower tolerogenic potential, resulting in a more robust CD8+ T cell activation and their functional activity as well as significantly lower induction of regulatory T cells. These superior parameters are attributed, at least in part, to the ability of BG-matured DCs to resist potential immunosuppressive and pro-tolerogenic activity of various tumor cell lysates, including melanoma, renal carcinoma and glioblastoma.

The value of MMP-9 for breast and non-small cell lung cancer patients' survival.

PURPOSE: Matrix metalloproteinases (MMPs) are implicated in cancer cells invasion and metastasis processes and have been investigated as potential cancer biomarkers. In this study MMP-9 gene expression and MMP-9 -1562 C/T polymorphism in breast and non-small cell lung cancer patients' blood and tumor samples and its correlation with clinicopathological parameters were investigated. MATERIAL/METHODS: MMP-9 gene expression was assessed by reverse transcription - polymerase chain reaction method in 108 cancer patients' blood and tumor samples. MMP-9 -1562 C/T polymorphism was determined by the polymerase chain reaction - based restriction fragment length polymorphism method. RESULTS: Significant relationship of MMP-9 gene expression and tumor differentiation grade was found only between groups with G1 and G3 breast tumors. Low survival rates were identified among positive MMP-9 expression in blood and ductal carcinoma of the breast (p=0.01) and negative progesterone receptor reaction (p=0.04). Significant differences in the distribution among genotypes were found between groups with stage I and stages III/IV (p=0.005) as well as between groups with lymph node status N0 and N1 (p<0.001). Breast cancer patients with tumor differentiation grade G3 and identified CC variant had a longer survival time (p=0.014). Shorter survival time was found among positive MMP-9 expression in tumor and stage I non-small cell lung cancer patients with negative lymph node (p=0.012) and squamous cell carcinoma (p=0.019). CONCLUSIONS: Expression of MMP-9 in blood and tumor together indicates worse prognosis for breast cancer patients.

A Shandon PapSpin liquid-based gynecological test: A split-sample and direct-to-vial test with histology follow-up study.

BACKGROUND: Studies for liquid-based Papanicolaou (Pap) tests reveal that liquid-based cytology (LBC) is a safe and effective alternative to the conventional Pap smear. Although there is research on ThinPrep and SurePath systems, information is lacking to evaluate the efficiency and effectiveness of systems based on cytocentrifugation. This study is designed to determine the sensitivity and specificity of the Shandon PapSpin (ThermoShandon, Pittsburgh, Pennsylvania, USA) liquid-based gynecological system. We used split-sample and direct-to-vial study design. MATERIALS AND METHODS: 2,945 women referred to prophylactic check-up were enrolled in this study. Split sample design was used in 1,500 women and residual cervical cytology specimen from all these cases was placed in fluid for PapSpin preparation after performing conventional smear. The direct-to-vial study was carried out in another cohort of 1,445 women in whom the entire cervical material was investigated using only the PapSpin technique. Follow up histological diagnoses for 141 women were obtained from both study arms following 189 abnormal cytology cases. 80 LBC cases from the split sample group and 61 LBC cases in the direct-to-vial group were correlated with the histology results. The sensitivity and secificity of the conventional smear and PapSpin tests in both study arms were compared. RESULTS: In the split sample group, conventional smears showed a higher proportion of ASC-US (atypical cells undetermined significance): 31 (2.1%) vs 10 (0.7%) in PapSpin (P = 0.001). A higher proportion of unsatisfactory samples was found in the conventional smear group: 25 (1.7%) vs 6 (0.4%) cases (P = 0.001). In the split sample group, the sensitivity of the conventional and PapSpin tests was 68.7% vs 78.1%, and the specificity 93.8% vs 91.8%, respectively. In the direct to vial group PapSpin sensitivity was 75.9% and specificity 96.5%. The differences in sensitivity and specificity were not significant. The positive predictive values for the conventional and PapSpin methods were not different in the split sample group: 88.0% vs 86.2% and 95.7% in the direct-to-vial group. Also, no differences were found for negative predictive value (82.1, 86.8% and 80.0% respectively). CONCLUSIONS: PapSpin showed good qualitative results in both study arms, even after the material splitting in the first study arm, and is a good alternative to the conventional Pap smear. Additionally, the PapSpin method offers several advantages such as the opportunity to prepare duplicate slides, option for HPV DNA testing and cell block preparations from residual material. Microscopic evaluation of thinner cell preparations is less time consuming than the conventional Pap smears.