Comparative Efficacy and Quality Characterizatin of Mayhaw (Crataegus Opaca) Juice Extraction.

The comparative efficacy and quality characterization of juice by six extraction methods utilizing either frozen or fresh \u27Texas Star\u27 mayhaw (Crataegus opaca Hook. and Arn.) fruit obtained from the 1998 and 1999 crop season were conducted according to temperature (cold-press or hot-press extraction) and fruit pulp pretreatment (whole or pulverized fruit with or without pectolytic enzyme). Extraction methods were steam extraction using whole fruit without added pulp juice (SE); steam extraction using whole fruit with added pulp juice (SEP); cold-press extraction using whole fruit (CPE); cold-press extraction using finely macerated fruit pulp (CPEG); hot-press extraction using finely macerated fruit pulp (HPEG), and hotpress extraction in combination with a pectolytic enzyme pretreatment (ENZ). Maceration of fruit, plus the application of heat to the fruit pulp prior to expression, had a significant influence in juice extraction efficacy for both fresh and frozen fruit. Application of a commercial pectolytic enzyme (0.20% w/w) to mayhaw pulp mash (1 hr at 32°C) increased extraction efficacy by of 21%. ENZ-fresh extraction had the highest percent total soluble solids (TSS) at 8.1% and was 7% more efficient in recovery as compared to ENZ-frozen extraction (7.6%). The fructose to glucose (F/G) ratio of juice recovered from all extraction methods utilizing either fresh or frozen fruit was greater than 2.0. In total sugars (fresh or frozen) there were no significant differences found between hot or cold-press extracted juices or the use of whole or macerated fruit. The CPEG extraction method using fresh fruit had a CIE b* color-value (2.55) that was significantly higher than all other methods of juice extraction. There were no significant differences in juice pH, which ranged from 2.9 to 3.1, between either fresh or frozen extracted juices. Juice from one cultivar of muscadine grape (Vitis rotundifolia Michx.) \u27Carlos\u27 (bronze-skinned) was mixed with varying levels of juice from \u27Texas Star\u27 mayhaw. Panelists\u27 mean scores collectively were favorable of either 60:40 or 40:60 mayhaw:muscadine juice blends and Taste contributed the most to overall acceptance

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival