Islet Harvest in Carbon Monoxide-Saturated Medium for Chronic Pancreatitis Patients Undergoing Islet Autotransplantation

Stresses encountered during human islet isolation lead to unavoidable beta-cell death after transplantation. This reduces the chance of insulin independence in chronic pancreatitis patients undergoing total pancreatectomy and islet autotransplantation. We tested whether harvesting islets in carbon monoxide-saturated solutions is safe and can enhance islet survival and insulin independence after total pancreatectomy and islet autotransplantation. Chronic pancreatitis patients who consented to the study were randomized into carbon monoxide (islets harvested in a carbon monoxide-saturated medium) or control (islets harvested in a normal medium) groups. Islet yield, viability, oxygen consumption rate, beta-cell death (measured by unmethylated insulin DNA), and serum cytokine levels were measured during the peri-transplantation period. Adverse events, metabolic phenotypes, and islet function were measured prior and at 6 months post-transplantation. No adverse events directly related to the infusion of carbon monoxide islets were observed. Carbon monoxide islets showed significantly higher viability before transplantation. Subjects receiving carbon monoxide islets had less beta-cell death, decreased CCL23, and increased CXCL12 levels at 1 or 3 days post transplantation compared with controls. Three in 10 (30%) of the carbon monoxide subjects and none of the control subjects were insulin independent. This pilot trial showed for the first time that harvesting human islets in carbon monoxide-saturated solutions is safe for total pancreatectomy and islet autotransplantation patients.Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Potato Ingestion as an Effective Race Fuel Alternative to Improve Cycling Performance in Trained Cyclists

Carbohydrate (CHO) ingestion is an established strategy to improve endurance performance. Race fuels should not only sustain performance, but also be readily digested and absorbed and replenish electrolytes. Potatoes are a cost-effective option that fulfills these criteria; however, their impact on endurance performance remains unexamined. PURPOSE: Compare the effects of potato purée (POT) ingestion during endurance cycling on subsequent performance versus commercial CHO gel (GEL) or a control (water, CTL). METHODS: Twelve trained cyclists (31±9y; 71±8kg; VO2max: 61±9mL/kg/min) consumed a standardized breakfast then performed a 2h cycling challenge (60-85%VO2max) followed by a time trial (6kJ/kg body mass) while consuming POT, GEL, or CTL in a randomized-crossover design. POT, GEL and CTL were administered with U-[13C6]glucose for an indirect estimate of gastric emptying rate. Repeated blood samples were collected. RESULTS: Time trial performance significantly improved (P=0.03) with POT (33.0±4.5min) and GEL (33.0±4.2min) versus CTL condition (39.5±7.9min); while POT and GEL conditions (P=1.00) had no difference. Post-challenge, blood glucose concentrations were lower (P0.05). CONCLUSION: Potatoes served as a viable alternative to commercial gels by sustaining performance and blood glucose concentrations during endurance cycling events in trained cyclists

Vaccination with poly(IC:LC) and peptide-pulsed autologous dendritic cells in patients with pancreatic cancer

Abstract Background Dendritic cells (DCs) enhance the quality of anti-tumor immune response in patients with cancer. Thus, we posit that DC-based immunotherapy, in conjunction with toll-like receptor (TLR)-3 agonist poly-ICLC, is a promising approach for harnessing immunity against metastatic or locally advanced unresectable pancreatic cancer (PC). Methods We generated autologous DCs from the peripheral blood of HLA-A2+ patients with PC. DCs were pulsed with three distinct A2-restricted peptides: 1) human telomerase reverse transcriptase (hTERT, TERT572Y), 2) carcinoembryonic antigen (CEA; Cap1-6D), and 3) survivin (SRV.A2). Patients received four intradermal injections of 1 × 107 peptide-pulsed DC vaccines every 2 weeks (Day 0, 14, 28, and 42). Concurrently, patients received intramuscular administration of Poly-ICLC at 30 μg/Kg on vaccination days (i.e., day 0, 14, 28, and 42), as well as on days 3, 17, 21, 31, 37, and 45. Our key objective was to assess safety and feasibility. The effect of DC vaccination on immune response was measured at each DC injection time point by enumerating the phenotype and function of patient T cells. Results Twelve patients underwent apheresis: nine patients with metastatic disease, and three patients with locally advanced unresectable disease. Vaccines were successfully manufactured from all individuals. We found that this treatment was well-tolerated, with the most common symptoms being fatigue and/or self-limiting flu-like symptoms. Among the eight patients who underwent imaging on day 56, four patients experienced stable disease while four patients had disease progression. The median overall survival was 7.7 months. One patient survived for 28 months post leukapheresis. MHC class I –tetramer analysis before and after vaccination revealed effective generation of antigen-specific T cells in three patients with stable disease. Conclusion Vaccination with peptide-pulsed DCs in combination with poly-ICLC is safe and induces a measurable tumor specific T cell population in patients with advanced PC. Trial registration NCT01410968 ; Name of registry: clinicaltrials.gov; Date of registration: 08/04/2011)

Additional file 1: Figure S1. of Vaccination with poly(IC:LC) and peptide-pulsed autologous dendritic cells in patients with pancreatic cancer

Cytokine secretion by DCs. Frozen DCs obtained from patients were thawed and cultured with GM-CSF/IL-4 overnight before adding poly (IC:LC) for maturation overnight. The supernatant was collected after 16 hrs. and evaluated for cytokines IL12 (upper panel), and IL10 (lower panel) as per the manufacturer’s protocol. Figure S2. PFS and OS details. The raw data showing details of progression free survival (PFS) and the median overall survival (OS) is presented in tabular form. Figure S3. Gating scheme for flow cytometry analysis. PBMC was obtained from patients post vaccination and cryopreserved cells were stained using the multiple fluorochrome-conjugated antibodies. Cells were gated based on singlets (FSC-A vs FSC-H), size (SSC vs FSC-H), a live-dead stain (L/D), and subsequently markers to determine specific cell phenotypes. A) CD3+ T cells were phenotyped for CD4 and CD8. B) CD19 B cells were identified. C) NK cells were identified based on their CD56 and CD16 expression. The data was acquired using BD FACS Aria and analyzed using FlowJo software. (PDF 351 kb

Comparative analysis of antibodies to SARS-CoV-2 between asymptomatic and convalescent patients

Summary: The SARS-CoV-2 viral pandemic has induced a global health crisis, which requires more in-depth investigation into immunological responses to develop effective treatments and vaccines. To understand protective immunity against COVID-19, we screened over 60,000 asymptomatic individuals in the Southeastern United States for IgG antibody positivity against the viral Spike protein, and approximately 3% were positive. Of these 3%, individuals with the highest anti-S or anti-RBD IgG level showed a strong correlation with inhibition of ACE2 binding and cross-reactivity against non-SARS-CoV-2 coronavirus S-proteins. We also analyzed samples from 94 SARS-CoV-2 patients and compared them with those of asymptomatic individuals. SARS-CoV-2 symptomatic patients had decreased antibody responses, ACE2 binding inhibition, and antibody cross-reactivity. Our study shows that healthy individuals can mount robust immune responses against SARS-CoV-2 without symptoms. Furthermore, IgG antibody responses against S and RBD may correlate with high inhibition of ACE2 binding in individuals tested for SARS-CoV-2 infection or post vaccination