STING-Dependent Cytosolic DNA Sensing Promotes Radiation-Induced Type I Interferon-Dependent Antitumor Immunity in Immunogenic Tumors.

Ionizing radiation-mediated tumor regression depends on type I interferon (IFN) and the adaptive immune response, but several pathways control I IFN induction. Here, we demonstrate that adaptor protein STING, but not MyD88, is required for type I IFN-dependent antitumor effects of radiation. In dendritic cells (DCs), STING was required for IFN-? induction in response to irradiated-tumor cells. The cytosolic DNA sensor cyclic GMP-AMP (cGAMP) synthase (cGAS) mediated sensing of irradiated-tumor cells in DCs. Moreover, STING was essential for radiation-induced adaptive immune responses, which relied on type I IFN signaling on DCs. Exogenous IFN-? treatment rescued the cross-priming by cGAS or STING-deficient DCs. Accordingly, activation of STING by a second messenger cGAMP administration enhanced antitumor immunity induced by radiation. Thus radiation-mediated antitumor immunity in immunogenic tumors requires a functional cytosolic DNA-sensing pathway and suggests that cGAMP treatment might provide a new strategy to improve radiotherapy.

4-Hydroxyacetophenone modulates the actomyosin cytoskeleton to reduce metastasis.

Metastases are the cause of the vast majority of cancer deaths. In the metastatic process, cells migrate to the vasculature, intravasate, extravasate, and establish metastatic colonies. This pattern of spread requires the cancer cells to change shape and to navigate tissue barriers. Approaches that block this mechanical program represent new therapeutic avenues. We show that 4-hydroxyacetophenone (4-HAP) inhibits colon cancer cell adhesion, invasion, and migration in vitro and reduces the metastatic burden in an in vivo model of colon cancer metastasis to the liver. Treatment with 4-HAP activates nonmuscle myosin-2C (NM2C) (MYH14) to alter actin organization, inhibiting the mechanical program of metastasis. We identify NM2C as a specific therapeutic target. Pharmacological control of myosin isoforms is a promising approach to address metastatic disease, one that may be readily combined with other therapeutic strategies.

Synthesis and Thermal Studies of Two Phosphonium Tetrahydroxidohexaoxidopentaborate(1-) Salts: Single-Crystal XRD Characterization of [iPrPPh3][B5O6(OH)4]·3.5H2O and [MePPh3][B5O6(OH)4]·B(OH)3·0.5H2O.

Two substituted phosphonium tetrahydoxidohexaoxidopentaborate(1-) salts, [iPrPPh3][B5O6(OH)4]·3.5H2O (1) and [MePPh3][B5O6(OH)4]·B(OH)3·0.5H2O (2), were prepared by templated self-assembly processes with good yields by crystallization from basic methanolic aqueous solutions primed with B(OH)3 and the appropriate phosphonium cation. Salts 1 and 2 were characterized by spectroscopic (NMR and IR) and thermal (TGA/DSC) analysis. Salts 1 and 2 were thermally decomposed in air at 800 °C to glassy solids via the anhydrous phosphonium polyborates that are formed at lower temperatures (<300 °C). BET analysis of the anhydrous and pyrolysed materials indicated they were non-porous with surface areas of 0.2-2.75 m2/g. Rhe recrystallization of 1 and 2 from aqueous solution afforded crystals suitable for single-crystal XRD analyses. The structure of 1 comprises alternating cationic/anionic layers with the H2O/pentaborate(1-) planes held together by H-bonds. The cationic planes have offset face-to-face (off) and vertex-to-face (vf) aromatic ring interactions with the iPr groups oriented towards the pentaborate(1-)/H2O layers. The anionic lattice in 2 is expanded by the inclusion of B(OH)3 molecules to accommodate the large cations; this results in the formation of a stacked pentaborate(1-)/B(OH)3 structure with channels occupied by the cations. The cations within the channels have vf, ef (edge-to-face), and off phenyl embraces. Both H-bonding and phenyl embrace interactions are important in stabilizing these two solid-state structures.

Structural (XRD) Characterization and an Analysis of H-Bonding Motifs in Some Tetrahydroxidohexaoxidopentaborate(1-) Salts of N-Substituted Guanidinium Cations.

The synthesis and characterization of six new substituted guanidium tetrahydroxidohexaoxidopentaborate(1-) salts are reported: [C(NH2)2(NHMe)][B5O6(OH)4]·H2O (1), [C(NH2)2(NH{NH2})][B5O6(OH)4] (2), [C(NH2)2(NMe2)][B5O6(OH)4] (3), [C(NH2)(NMe2)2][B5O6(OH)4] (4), [C(NHMe)(NMe2)2][B5O6(OH)4]·B(OH)3 (5), and [TBDH][B5O6(OH)4] (6) (TBD = 1,5,7-triazabicyclo [4.4.0]dec-5-ene). Compounds 1-6 were prepared as crystalline salts from basic aqueous solution via self-assembly processes from B(OH)3 and the appropriate substituted cation. Compounds 1-6 were characterized by spectroscopic (NMR and IR) and by single-crystal XRD studies. A thermal (TGA) analysis on compounds 1-3 and 6 demonstrated that they thermally decomposed via a multistage process to B2O3 at >650 °C. The low temperature stage (<250 °C) was endothermic and corresponded to a loss of H2O. Reactant stoichiometry, solid-state packing, and H-bonding interactions are all important in assembling these structures. An analysis of H-bonding motifs in known unsubstituted guanidinium salts [C(NH2)3]2[B4O5(OH)4]·2H2O, [C(NH2)3][B5O6(OH)4]·H2O, and [C(NH2)3]3[B9O12(OH)6] and in compounds 1-6 revealed that two important H-bonding R22(8) motifs competed to stabilize the observed structures. The guanidinium cation formed charge-assisted pincer cation-anion H-bonded rings as a major motif in [C(NH2)3]2[B4O5(OH)4]·2H2O and [C(NH2)3]3[B9O12(OH)6], whereas the anion-anion ring motif was dominant in [C(NH2)3][B5O6(OH)4]·H2O and in compounds 1-6. This behaviour was consistent with the stoichiometry of the salt and packing effects also strongly influencing their solid-state structures.

Recent Advances in Crystalline Oxidopolyborate Complexes of d-Block or p-Block Metals: Structural Aspects, Syntheses, and Physical Properties.

Crystalline materials containing hybrid inorganic-organic metal borates (complexes with oxidoborate ligands) display a variety of novel framework building blocks. The structural aspects of these hybrid metallaoxidoborates containing Cd(II), Co(II), Cu(II), Ga(III), In(III), Mn(II), Ni(II) or Zn(II) metal centers are discussed in this review. The review describes synthetic approaches to these hybrid materials, their physical properties, their spectroscopic properties and their potential applications.

Pentaborate(1-) Salts and a Tetraborate(2-) Salt Derived from C2- or C3-Linked Bis(alkylammonium) Dications: Synthesis, Characterization, and Structural (XRD) Studies.

The synthesis of a number of pentaborate(1-) salts from cations arising from N-substituted α,α-, α,ß-, and α,γ-diaminoalkanes has been attempted in aqueous solution from B(OH)3 and the appropriate diammine in a 10:1 ratio. Despite relatively mild work-up conditions the pentaborate(1-) salts prepared were not always as anticipated and the following compounds were isolated in good yield: [Me2NH(CH2)2NHMe2][B5O6(OH)4]2 (1), [Et2NH(CH2)2NHEt2][B5O6(OH)4]2 (2), [Et2NH2][B5O6(OH)4] (3), [Me2NH2][B5O6(OH)4] (4), [Me2NH(CH2)3NHMe2][B5O6(OH)4]2 (5), [Et2NH(CH2)3NHEt2][B5O6(OH)4]2 (6), [Me3NCH2CH=CH2][B5O6(OH)4] (7), and [Me3N(CH2)3NMe3] [B5O6(OH)4]2.0.5H2O (8). The tetraborate(2-) salt, [Me3N(CH2)2NMe3][B4O5(OH)4].2B(OH)3.2H2O (9) was obtained in moderate yield (41%) from a 3:1 reaction of B(OH)3 with [Me3N(CH2)2NMe3](OH)2. All compounds were characterized by spectroscopy (1H, 11B, 13C NMR and IR) and thermal gravimetric analysis (TGA). BET analysis on materials derived thermally from selected samples (1, 2, 6, 7) all had porosities of < 1 m2/g, demonstrating that they were non-porous. Single-crystal XRD structures were obtained for 2, 3, 7, 8 and 9 and all contain extensive H-bonded polyborate lattices.

Patient Relations Measurement and Reporting to Improve Quality and Safety: Lessons from a Pilot Project.

BACKGROUND: Effective patient relations can improve the patient experience and the safe delivery of care. Recent Ontario policy and legislative changes have increased accountabilities for healthcare organizations and expanded Health Quality Ontario's mandate to measure and report on patient relations. The purpose of this pilot project was to support improved care by validating standardized measures, data submission processes and prototype reporting of patient relations indicators for the hospital, home and community care and long-term care sectors across Ontario. METHODS: Health Quality Ontario identified potential indicators and best practices by performing a comprehensive environmental scan and consulting with experts, including patients and caregivers. It shortlisted indicators based on alignment to best practices and Ontario legislative requirements. A provincial advisory group then used a modified Delphi process to prioritize and recommend five patient relations indicators for province-wide measurement and comparative public reporting. Through the pilot project, these indicators were validated using facility-level data for fiscal year (FY) 2015-2016 from 29 hospitals, home and community care organizations and long-term care homes across Ontario. RESULTS: In June 2016, Health Quality Ontario recruited 34 organizations for the pilot project. Twenty-nine sites successfully submitted summary-level data on patient relations indicators. More than 90% of the required data were retrieved from existing papers or electronic systems. All sites mapped facility-level "complaint" and "action taken" categories to the provincial standardized categories. Across the three health sectors, "care and treatment" was the top complaint category in FY 2015-2016. CONCLUSIONS: This pilot project reinforced the value of measuring patient relations and reporting feedback to support facility- and system-level improvement. The pilot sites and provincial advisory group recommended phased implementation. This would permit healthcare organizations to standardize data collection and align with provincial indicators and categories. The next step would be voluntary data submission to Health Quality Ontario in advance of any reporting. To facilitate voluntary implementation, Health Quality Ontario included one indicator, "complaints acknowledged," in the annual Quality Improvement Plans beginning in FY 2018-2019. This will allow organizations to monitor and report on the percentage of complaints acknowledged within 2, 5 and 10 days. Implementation will evolve based on input from patients, health sector organizations, Local Health Integration Networks and the Patient Ombudsman.

Comparative Analysis of the Antimicrobial Activities of Plant Defensin-Like and Ultrashort Peptides against Food-Spoiling Bacteria.

UNLABELLED: Antimicrobial peptides offer potential as novel therapeutics to combat food spoilage and poisoning caused by pathogenic and nonpathogenic bacteria. Our previous studies identified the peptide human beta-defensin 3 (HBD3) as a potent antimicrobial agent against a wide range of beer-spoiling bacteria. Thus, HBD3 is an excellent candidate for development as an additive to prevent food and beverage spoilage. To expand the repertoire of peptides with antimicrobial activity against bacteria associated with food spoilage and/or food poisoning, we carried out an in silico discovery pipeline to identify peptides with structure and activity similar to those of HBD3, focusing on peptides of plant origin. Using a standardized assay, we compared the antimicrobial activities of nine defensin-like plant peptides to the activity of HBD3. Only two of the peptides, fabatin-2 and Cp-thionin-2, displayed antimicrobial activity; however, the peptides differed from HBD3 in being sensitive to salt and were thermostable. We also compared the activities of several ultrashort peptides to that of HBD3. One of the peptides, the synthetic tetrapeptide O3TR, displayed biphasic antimicrobial activity but had a narrower host range than HBD3. Finally, to determine if the peptides might act in concert to improve antimicrobial activity, we compared the activities of the peptides in pairwise combinations. The plant defensin-like peptides fabatin-2 and Cp-thionin-2 displayed a synergistic effect with HBD3, while O3TR was antagonistic. Thus, some plant defensin-like peptides are effective antimicrobials and may act in concert with HBD3 to control bacteria associated with food spoilage and food poisoning. IMPORTANCE: Food spoilage and food poisoning caused by bacteria can have major health and economic implications for human society. With the rise in resistance to conventional antibiotics, there is a need to identify new antimicrobials to combat these outbreaks in our food supply. Here we screened plant peptide databases to identify peptides that share structural similarity with the human defensin peptide HBD3, which has known antimicrobial activity against food-spoiling bacteria. We show that two of the plant peptides display antimicrobial activity against bacteria associated with food spoilage. When combined with HBD3, the peptides are highly effective. We also analyzed the activity of an easily made ultrashort synthetic peptide, O3TR. We show that this small peptide also displays antimicrobial activity against food-spoiling bacteria but is not as effective as HBD3 or the plant peptides. The plant peptides identified are good candidates for development as natural additives to prevent food spoilage.

A new decaoxidooctaborate(2-) anion, [B8O10(OH)6](2-): synthesis and characterization of [Co(en)3][B5O6(OH)4][B8O10(OH)6)]·5H2O (en = 1,2-diaminoethane).

The synthesis and X-ray diffraction structure of [Co(en)3][B5O6(OH)4][B8O10(OH)6]·5H2O (1) are reported. Compound 1 arises through a selective-templating process from a Dynamic Combinatorial Library of polyborate anions. Compound 1 contains two different polyborate species, with [B8O10(OH)6](2-) being particularly novel. It is comprised of fused tetraborate and pentaborate anions with a 4-coordinate B atom and a 3-coordinate O atom in common.

Radiation-induced equilibrium is a balance between tumor cell proliferation and T cell-mediated killing.

Local failures following radiation therapy are multifactorial, and the contributions of the tumor and the host are complex. Current models of tumor equilibrium suggest that a balance exists between cell birth and cell death due to insufficient angiogenesis, immune effects, or intrinsic cellular factors. We investigated whether host immune responses contribute to radiation-induced tumor equilibrium in animal models. We report an essential role for immune cells and their cytokines in suppressing tumor cell regrowth in two experimental animal model systems. Depletion of T cells or neutralization of IFN-γ reversed radiation-induced equilibrium, leading to tumor regrowth. We also demonstrate that PD-L1 blockade augments T cell responses, leading to rejection of tumors in radiation-induced equilibrium. We identify an active interplay between tumor cells and immune cells that occurs in radiation-induced tumor equilibrium and suggest a potential role for disruption of the PD-L1/PD-1 axis in increasing local tumor control.

A meta-analysis of prospective studies on the role of physical activity and the prevention of Alzheimer's disease in older adults.

BACKGROUND: The incidence of Alzheimer's disease is increasing as the global population ages. Given the limited success of pharmaceuticals in preventing this disease, a greater emphasis on non-pharmaceutical approaches is needed. The aim of this study was to quantify the association between Alzheimer's disease and physical activity in older adults over the age of 65 years. METHODS: A meta-analytic approach was used to determine if physical activity reduced the risk of Alzheimer's disease in individuals 65 years or older. Some evidence indicates that physical activity may improve cognitive function in older adults, while other evidence is inconclusive. The purpose of this study was to examine if prevention of Alzheimer's disease is possible if started at a later age. The precise brain changes that occur with the onset of Alzheimer's disease are not fully known, and therefore may still be influenced by preventative measures even in advancing age. Determining if physical activity can inhibit the onset of the disease at any age may motivate individuals to adopt an "it's never too late" mentality on preventing the onset of this debilitating disease. Longitudinal studies of participants who were 65 years or older at baseline were included. A total of 20,326 participants from nine studies were included in this analysis. RESULTS: The fixed effects risk ratio is estimated as 0.61 (95% CI 0.52-0.73) corresponding to a statistically significant overall reduction in risk of Alzheimer's disease in physically active older adults compared to their non-active counterparts. CONCLUSION: Physical activity was associated with a reduced risk of Alzheimer's disease in adults over the age of 65 years. Given the limited treatment options, greater emphasis should be paid to primary prevention through physical activity amongst individuals at high-risk of Alzheimer's disease, such as those with strong genetic and family history.

Patient and procedure-specific risk factors for deep infection after primary shoulder arthroplasty.

BACKGROUND: Deep infection after shoulder arthroplasty is a diagnostic and therapeutic challenge. The current literature on this topic is from single institutions or Medicare samples, lacking generalizability to the larger shoulder arthroplasty population. QUESTIONS/PURPOSES: We sought to identify (1) patient-specific risk factors for deep infection, and (2) the pathogen profile after primary shoulder arthroplasty in a large integrated healthcare system. METHODS: A retrospective cohort study was conducted. Of 4528 patients identified, 320 had died and 302 were lost to followup. The remaining 3906 patients had a mean followup of 2.7 years (1 day-7 years). The study endpoint was the diagnosis of deep infection, which was defined as revision surgery for infection supported clinically by more than one of the following criteria: purulent drainage from the deep incision, fever, localized pain or tenderness, a positive deep culture, and/or a diagnosis of deep infection made by the operating surgeon based on intraoperative findings. Risk factors evaluated included age, sex, race, BMI, diabetes status, American Society for Anesthesiologists (ASA) score, traumatic versus elective procedure, and type of surgical implant. For patients with deep infections, we reviewed the surgical notes and microbiology records for the pathogen profile. Multivariable Cox regression models were used to evaluate the association of risk factors and deep infection. Adjusted hazard ratios and 95% CI are presented. RESULTS: With every 1-year increase in age, a 5% (95% CI, 2%-8%) lower risk of infection was observed. Male patients had a risk of infection of 2.59 times (95% CI, 1.27-5.31) greater than female patients. Patients undergoing primary reverse total shoulder arthroplasty had a 6.11 times (95% CI, 2.65-14.07) greater risk of infection compared with patients having primary unconstrained total shoulder arthroplasty. Patients having traumatic arthroplasties were 2.98 times (95% CI, 1.15-7.74) more likely to have an infection develop than patients having elective arthroplasties. BMI, race, ASA score, and diabetes status were not associated with infection risk (all p > 0.05). Propionibacterium acnes was the most commonly cultured organism, accounting for 31% of isolates. CONCLUSIONS: Younger, male patients are at greater risk for deep infection after primary shoulder arthroplasty. Reverse total shoulder arthroplasty and traumatic shoulder arthroplasties also carry a greater risk for infection. Propionibacterium acnes was the most prevalent pathogen causing infection in our primary shoulder arthroplasty population. LEVEL OF EVIDENCE: Level II, prognostic study. See the Instructions for Authors for a complete description of levels of evidence.

Co(II)-doped hybrid Zn(II) tetraborate complexes, [ZnxCo(1-x)(1,3-dap)B4O7] (1,3-dap = 1,3-diaminopropane): BET analysis and N2/H2O/D2O adsorption studies.

A series of mono/bimetallic isostructural hybrid tetraborates of the general formula [ZnxCo(1-x)(1,3-dap)B4O7] has been prepared using a solvothermal method. Their adsorption/desorption curves for H2O and D2O demonstrate that these materials have a stronger affinity for H2O than for D2O and enrich the D2O content of D2O/H2O mixtures.

MUC1-induced alterations in a lipid metabolic gene network predict response of human breast cancers to tamoxifen treatment.

The mucin 1 (MUC1) oncoprotein is aberrantly overexpressed in human breast cancers. Although MUC1 modulates the activity of estrogen receptor alpha (ER), there is no information regarding the effects of MUC1 on global gene expression patterns and the potential role of MUC1-induced genes in predicting outcome for breast cancer patients. We have developed an experimental model of MUC1-induced transformation that has identified the activation of genes involved in cholesterol and fatty acid metabolism. A 38-gene set of experimentally derived MUC1-induced genes associated with lipid metabolism was applied to the analysis of ER(+) breast cancer patients treated with tamoxifen. The results obtained from 2 independent databases demonstrate that patients overexpressing MUC1 and the lipid metabolic pathways are at significantly higher risk for death and recurrence/distant metastasis. By contrast, these genes were not predictive in untreated patients. Furthermore, a positive correlation was found between expression of the 38-gene set and the ER signaling pathway. These findings indicate that (i) MUC1 regulates cholesterol and fatty acid metabolism, and (ii) activation of these pathways in ER(+) breast cancers predicts failure to tamoxifen treatment.

Rapid disinfection of radiology treatment rooms using an autonomous ultraviolet germicidal irradiation robot.

BACKGROUND: Ultraviolet germicidal irradiation (UVGI) technologies have emerged as a promising adjunct to manual cleaning, however, their potential to shorten cleaning times remains unexplored. METHODS: A <10-minute disinfection procedure was developed using a robotic UVGI platform. The efficacy and time to perform the UVGI procedure in a CT scan treatment room was compared with current protocols involving manual disinfection using biocides. For each intervention, environmental samples were taken at 12 locations in the room before and after disinfection on seven distinct occasions. RESULTS: The mean UVC dose at each sample location was found to be 13.01 ± 4.36 mJ/cm2, which exceeded published UVC thresholds for achieving log reductions of many common pathogens. Significant reductions in microbial burden were measured after both UVGI (P≤.001) and manual cleaning (P≤.05) conditions, with the UVGI procedure revealing the largest effect size (r = 0.603). DISCUSSION: These results support the hypothesis that automated deployments of UVGI technology can lead to germicidal performance that is comparable with, and potentially better than, current manual cleaning practices. CONCLUSIONS: Our findings provide early evidence that the incorporation of automated UVGI procedures into cleaning workflow could reduce turnaround times in radiology, and potentially other hospital settings.

Reciprocally rewiring and repositioning the Integration Host Factor (IHF) subunit genes in Salmonella enterica serovar Typhimurium: impacts on physiology and virulence.

The Integration Host Factor (IHF) is a heterodimeric nucleoid-associated protein that plays roles in bacterial nucleoid architecture and genome-wide gene regulation. The ihfA and ihfB genes encode the subunits and are located 350 kbp apart, in the Right replichore of the Salmonella chromosome. IHF is composed of one IhfA and one IhfB subunit. Despite this 1 : 1 stoichiometry, MS revealed that IhfB is produced in 2-fold excess over IhfA. We re-engineered Salmonella to exchange reciprocally the protein-coding regions of ihfA and ihfB, such that each relocated protein-encoding region was driven by the expression signals of the other's gene. MS showed that in this 'rewired' strain, IhfA is produced in excess over IhfB, correlating with enhanced stability of the hybrid ihfB-ihfA mRNA that was expressed from the ihfB promoter. Nevertheless, the rewired strain grew at a similar rate to the wild-type and was similar in competitive fitness. However, compared to the wild-type, it was less motile, had growth-phase-specific reductions in SPI-1 and SPI-2 gene expression, and was engulfed at a higher rate by RAW macrophage. Our data show that while exchanging the physical locations of its ihf genes and the rewiring of their regulatory circuitry are well tolerated in Salmonella, genes involved in the production of type 3 secretion systems exhibit dysregulation accompanied by altered phenotypes.

Therapeutic effects of ablative radiation on local tumor require CD8+ T cells: changing strategies for cancer treatment.

Patients with locally advanced cancer or distant metastasis frequently receive prolonged treatment with chemotherapy and/or fractionated radiotherapy (RT). Despite the initial clinical response, treatment resistance frequently develops and cure in these patients is uncommon. Developments in RT technology allow for the use of high-dose (or ablative) RT to target local tumors, with limited damage to the surrounding normal tissue. We report that reduction of tumor burden after ablative RT depends largely on T-cell responses. Ablative RT dramatically increases T-cell priming in draining lymphoid tissues, leading to reduction/eradication of the primary tumor or distant metastasis in a CD8(+) T cell-dependent fashion. We further demonstrate that ablative RT-initiated immune responses and tumor reduction are abrogated by conventional fractionated RT or adjuvant chemotherapy but greatly amplified by local immunotherapy. Our study challenges the rationale for current RT/chemotherapy strategies and highlights the importance of immune activation in preventing tumor relapse. Our findings emphasize the need for new strategies that not only reduce tumor burden but also enhance the role of antitumor immunity.

Synthesis and structural characterization of an unprecedented nonmetal cation polyborate salt containing two different "isolated" polyborate anions: [H2en]2[B4O5(OH)4][B7O9(OH)5]·3H2O (en = H2NCH2CH2NH2).

The nonmetal cation polyborate salt of stoichiometry [H(2)en](2)[B(11)O(18)(OH)]·7H(2)O is obtained from the reaction of 1,2-diaminoethane and boric acid (1:5 ratio) in H(2)O/MeOH. An X-ray crystallographic study of the product reveals that the polyborate moiety is composed of two isolated hydrated polyborate anions: [B(4)O(5)(OH)(4)](2-) and [B(7)O(9)(OH)(5)](2-). The structure is templated by the cations with the anions forming a supramolecular H-bonded network, augmented by additional H-bonds involving the waters of crystallization and the cations.

Ad.Egr-TNF and local ionizing radiation suppress metastases by interferon-beta-dependent activation of antigen-specific CD8+ T cells.

Ad.Egr-TNF is a radioinducible adenovector currently in phase 3 trials for inoperable pancreatic cancer. The combination of Ad.Egr-TNF and ionizing radiation (IR) contributes to local tumor control through the production of tumor necrosis factor-alpha (TNFalpha) in the tumor microenvironment. Moreover, clinical and preclinical studies with Ad.Egr-TNF/IR have suggested that this local approach suppresses the growth of distant metastatic disease; however, the mechanisms responsible for this effect remain unclear. These studies have been performed in wild-type (WT) and TNFR1,2(-/-) mice to assess the role of TNFalpha-induced signaling in the suppression of draining lymph node (DLN) metastases. The results demonstrate that production of TNFalpha in the tumor microenvironment induces expression of interferon (IFNbeta). In turn, IFNbeta stimulates the production of chemokines that recruit CD8(+) T cells to the tumor. The results further demonstrate that activation of tumor antigen-specific CD8(+) CTLs contributes to local antitumor activity and suppression of DLN metastases. These findings support a model in which treatment of tumors with Ad.Egr-TNF and IR is mediated by local and distant immune-mediated antitumor effects that suppress the development of metastases.