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

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Changes made from previous registration

The small pilot indicated the participants could complete the entire study within 45 minutes. Committee approved plan with no changes to materials (except adding the phrase "during sex" in two vignettes), including the outcome items. Reliability analyses for the outcome measure have not been conducted prior to beginning the full study. Distributions of items appeared to not indicate any response censoring (e..g, ceiling effects)

Monitoring the extent of vertical and lateral movement of human decomposition products through sediment using cholesterol as a biomarker

Due to the lack of human decomposition research facilities available in different geographical regions, the extent of movement of human decomposition products from a cadaver into various sedimentary environments, in different climates, has not been able to be studied in detail. In our study, a human cadaver was placed on the surface of a designated plot at the Australian Facility for Taphonomic Experimental Research (AFTER), the only human decomposition facility in Australia, where the natural process of decomposition was allowed to progress over 14 days in the Australian summer. Sediment columns (approximately 1 m deep) were collected at lateral distances of 0.25 m, 0.5 m, 1.0 m and 2.5 m in each of four directions from the centre of the torso. Plot elevation and weather data were also collected. Each sediment column was subdivided, dried and homogenised. A sample was isolated from each sediment subdivision, extracted with hexane, and the hexane extract cleaned with citrate buffer (pH 3), filtered and spiked with cholesterol-D 7 internal standard. After derivatisation with BSTFA + 1% TMCS, cholesterol was monitored in the samples using targeted gas chromatography tandem mass spectrometry analysis. A positive result for decomposition products was given if the cholesterol abundance in the test sample was higher than that detected in the \u27control\u27 samples of a similar substrate type collected prior to cadaver placement. Within the confines of the experimental design and the measured parameters, lateral leaching was observed over distances of up to 2.5 m from the centre of the torso, which was the maximum distance tested in the study. Vertical leaching was detected to depths of up to 49 cm below the ground surface. Such data can aid the development of policies related to plot sizing and sediment renewal and regeneration at other human decomposition facilities and at cemeteries. The density and distribution of cholesterol surrounding the cadaver in this study can also help forensic investigators interpret cases involving remains that have been moved or scavenged

Cytokines, Chemokines, and Growth Factors in Banked Human Donor Milk for Preterm Infants

Background: There has been a recent increase in availability of banked donor milk for feeding of preterm infants. This milk is pooled from donations to milk banks from carefully screened lactating women. The milk is then pasteurized by the Holder method to remove all microbes. The processed milk is frozen, banked, and sold to neonatal intensive care units (NICUs). The nutrient bioavailability of banked donor milk has been described, but little is known about preservation of immune components such as cytokines, chemokines, and growth factors (CCGF). Objective: The objective was to compare CCGF in banked donor milk with mother’s own milk (MOM). Methods: Aliquots (0.5 mL) were collected daily from MOM pumped by 45 mothers of NICU-admitted infants weighing \u3c 1500 grams at birth. All daily aliquots of each mother’s milk were pooled each week during 6 weeks of an infant’s NICU stay or for as long as the mother provided MOM. The weekly pooled milk was measured for a panel of CCGF through multiplexing using magnetic beads and a MAGPIX instrument. Banked donor milk samples (n = 25) were handled and measured in the same way as MOM. Results: Multiplex analysis revealed that there were levels of CCGF in banked donor milk samples comparable to values obtained from MOM after 6 weeks of lactation. Conclusion: These data suggest that many important CCGF are not destroyed by Holder pasteurization

<b>Postprandial leucine and insulin responses and toxicological effects of a novel whey protein hydrolysate-based supplement in rats</b>

<p><b>Abstract</b></p> <p>The purpose of this study was: aim 1) compare insulin and leucine serum responses after feeding a novel hydrolyzed whey protein (WPH)-based supplement versus a whey protein isolate (WPI) in rats during the post-absorptive state, and aim 2) to perform a thorough toxicological analysis on rats that consume different doses of the novel WPH-based supplement over a 30-day period. In male Wistar rats (~250 g, n = 40), serum insulin and leucine concentrations were quantified up to 120 min after one human equivalent dose of a WPI or the WPH-based supplement. In a second cohort of rats (~250 g, n = 20), we examined serum/blood and liver/kidney histopathological markers after 30 days of feeding low (1human equivalent dose), medium (3 doses) and high (6 doses) amounts of the WPH-based supplement. In aim 1, higher leucine levels existed at 15 min after WPH vs. WPI ingestion (p = 0.04) followed by higher insulin concentrations at 60 min (p = 0.002). In aim 2, liver and kidney histopathology/toxicology markers were not different 30 days after feeding with low, medium, high dose WPH-based supplementation or water only. There were no between-condition differences in body fat or lean mass or circulating clinical chemistry markers following the 30-day feeding intervention in aim 2. In comparison to WPI, acute ingestion of a novel WPH-based supplement resulted in a higher transient leucine response with a sequential increase in insulin. Furthermore, chronic ingestion of the tested whey protein hydrolysate supplement appears safe.</p