Role Of Il-27 And Tcr Stimulation In Inhibitory Receptor Expression

The adaptive immune response is necessary for control of pathogen burden in a wide range of infections. However, in the absence of active regulatory mechanisms, this protective response can lead to immune pathology. The cytokine interleukin (IL)-27 is required for control of exaggerated immune responses during toxoplasmosis and other infections and autoimmune settings. Multiple regulatory pathways have been found to be controlled by IL-27. However, there are gaps in our knowledge of the mechanisms by which IL-27 limits T cell responses. The present work focuses on the ability of IL-27 to promote expression of inhibitory receptors on T cells. The studies presented here establish that TCR and cytokines have distinct and complementary roles in promoting inhibitory receptor expression. In vitro, IL-27, type I IFN, and IFN-g induced expression of PD-L1 and Sca-1 on naïve murine CD4+ and CD8+ T cells in the absence of TCR stimulation. TCR stimulation induced expression of multiple inhibitory receptors and IL- 27 combined synergistically with TCR stimulation to further upregulate Ly6C, LAG-3, CTLA-4, TIGIT and TIM-3. This IL-27-mediated inhibitory expression was STAT1- dependent. The response to TCR stimulation was graded and thus a stronger TCR stimulus resulted in greater inhibitory receptor expression. In vivo, during infection with Toxoplasma gondii, IL-27 was required for full expression of Ly6C, PD-L1, LAG-3, CTLA-4, and TIGIT by parasite-specific T cells in the lung, a local site of infection, but not in the spleen. STAT1 was also required for full expression of LAG-3, CTLA-4, and TIGIT at local sites of infection. Taken together, these studies suggest a model in which inhibitory receptor expression on T cells is a graded regulatory pathway that is upregulated by exposure to increasing levels of TCR stimulation and cytokines present at sites of inflammation

Ecological pleiotropy and indirect effects alter the potential for evolutionary rescue

Invading predators can negatively affect naïve prey populations due to a lack of evolved defenses. Many species therefore may be at risk of extinction due to overexploitation by exotic predators. Yet the strong selective effect of predation might drive evolution of imperiled prey toward more resistant forms, potentially allowing the prey to persist. We evaluated the potential for evolutionary rescue in an imperiled prey using Gillespie eco‐evolutionary models (GEMs). We focused on a system parameterized for protists where changes in prey body size may influence intrinsic rate of population growth, space clearance rate (initial slope of the functional response), and the energetic benefit to predators. Our results show that the likelihood of rescue depends on (a) whether multiple parameters connected to the same evolving trait (i.e., ecological pleiotropy) combine to magnify selection, (b) whether the evolving trait causes negative indirect effects on the predator population by altering the energy gain per prey, (c) whether heritable trait variation is sufficient to foster rapid evolution, and (d) whether prey abundances are stable enough to avoid very rapid extinction. We also show that when evolution fosters rescue by increasing the prey equilibrium abundance, invasive predator populations also can be rescued, potentially leading to additional negative effects on other species. Thus, ecological pleiotropy, indirect effects, and system dynamics may be important factors influencing the potential for evolutionary rescue for both imperiled prey and invading predators. These results suggest that bolstering trait variation may be key to fostering evolutionary rescue, but also that the myriad direct and indirect effects of trait change could either make rescue outcomes unpredictable or, if they occur, cause rescue to have side effects such as bolstering the populations of invasive species

Genetic diversity affects the daily transcriptional oscillations of marine microbial populations

Marine microbial communities are genetically diverse but have robust synchronized daily transcriptional patterns at the genus level that are similar across a wide variety of oceanic regions. We developed a microarray-inspired gene-centric approach to resolve transcription of closely-related but distinct strains/ecotypes in high-throughput sequence data. Applying this approach to the existing metatranscriptomics datasets collected from two different oceanic regions, we found unique and variable patterns of transcription by individual taxa within the abundant picocyanobacteria Prochlorococcus and Synechococcus, the alpha Proteobacterium Pelagibacter and the eukaryotic picophytoplankton Ostreococcus. The results demonstrate that marine microbial taxa respond differentially to variability in space and time in the ocean. These intra-genus individual transcriptional patterns underlie whole microbial community responses, and the approach developed here facilitates deeper insights into microbial population dynamics

MEK inhibitors cobimetinib and trametinib, regressed a gemcitabine-resistant pancreatic-cancer patient-derived orthotopic xenograft (PDOX).

A pancreatic ductal adenocarcinoma (PDAC), obtained from a patient, was grown orthotopically in the pancreatic tail of nude mice to establish a patient-derived orthotopic (PDOX) model. Seven weeks after implantation, PDOX nude mice were divided into the following groups: untreated control (n = 7); gemcitabine (100 mg/kg, i.p., once a week for 2 weeks, n = 7); cobimetinib (5 mg/kg, p.o., 14 consecutive days, n = 7); trametinib (0.3 mg/kg, p.o., 14 consecutive days, n = 7); trabectedin (0.15 mg/kg, i.v., once a week for 2 weeks, n = 7); temozolomide (25 mg/kg, p.o., 14 consecutive days, n = 7); carfilzomib (2 mg/kg, i.v., twice a week for 2 weeks, n = 7); bortezomib (1 mg/kg, i.v., twice a week for 2 weeks, n = 7); MK-1775 (20 mg/kg, p.o., 14 consecutive days, n = 7); BEZ-235 (45 mg/kg, p.o., 14 consecutive days, n = 7); vorinostat (50 mg/kg, i.p., 14 consecutive days, n = 7). Only the MEK inhibitors, cobimetinib and trametinib, regressed tumor growth, and they were more significantly effective than other therapies (p < 0.0001, respectively), thereby demonstrating the precision of the PDOX models of PDAC and its potential for individualizing pancreatic-cancer therapy

Fluorescent humanized anti-CEA antibody specifically labels metastatic pancreatic cancer in a patient-derived orthotopic xenograft (PDOX) mouse model.

Pancreatic cancer is a highly lethal disease in part due to incomplete tumor resection. Targeting by tumor-specific antibodies conjugated with a fluorescent label can result in selective labeling of cancer in vivo for surgical navigation. In the present study, we describe a patient-derived orthotopic xenograft model of pancreatic cancer that recapitulated the disease on a gross and microscopic level, along with physiologic clinical manifestations. We additionally show that the use of an anti-CEA antibody conjugated to the near-infrared (NIR) fluorescent dye, IRDye800CW, can selectively highlight the pancreatic cancer and its metastases in this model with a tumor-to-background ratio of 3.5 (SEM 0.9). The present results demonstrate the clinical potential of this labeling technique for fluorescence-guided surgery of pancreatic cancer

Tumor-targeting Salmonella typhimurium A1-R in combination with doxorubicin eradicate soft tissue sarcoma in a patient-derived orthotopic xenograft (PDOX) model.

A patient with high grade undifferentiated pleomorphic soft-tissue sarcoma from a striated muscle was grown orthotopically in the right biceps femoris muscle of mice to establish a patient-derived orthotopic xenograft (PDOX) model. Twenty PDOX mice were divided into 4 groups: G1, control without treatment; G2, Salmonella typhimurium (S. typhimurium)A1-R administered by intratumoral (i.t.) injection once a week for 4 weeks; G3, doxorubicin (DOX) administered by intraperitoneal (i.p.) injection once a week for 4 weeks; G4, S. typhimurium A1-R (i.t.) administered once a week for 2 weeks followed by i.p. doxorubicin once a week for 2 weeks. On day 25 from the initiation of treatment, tumor volume in G2, G3, and G4 was significantly lower than G1. Mice found without gross tumor included one mouse (20%) in G2; one mouse (20%) in G3; and 3 mice (60%) in G4. Body weight loss did not significantly differ between the 3 treated groups or from the untreated control. Histological examination revealed eradication of tumor only in G4 where mice were treated with S. typhimurium A1-R followed by DOX. Our present study indicates future clinical potential of combining S. typhimurium A1-R with chemotherapy such as DOX for soft tissue sarcoma patients

Recombinant methioninase effectively targets a Ewing's sarcoma in a patient-derived orthotopic xenograft (PDOX) nude-mouse model.

Methionine dependence is due to the overuse of methionine for aberrant transmethylation reactions in cancer. Methionine dependence may be the only general metabolic defect in cancer. In order to exploit methionine dependence for therapy, our laboratory previously cloned L-methionine α-deamino-γ-mercaptomethane lyase [EC 4.4.1.11]). The cloned methioninase, termed recombinant methioninase, or rMETase, has been tested in mouse models of human cancer cell lines. Ewing's sarcoma is recalcitrant disease even though development of multimodal therapy has improved patients'outcome. Here we report efficacy of rMETase against Ewing's sarcoma in a patient-derived orthotopic xenograft (PDOX) model. The Ewing's sarcoma was implanted in the right chest wall of nude mice to establish a PDOX model. Eight Ewing's sarcoma PDOX mice were randomized into untreated control group (n = 4) and rMETase treatment group (n = 4). rMETase (100 units) was injected intraperitoneally (i.p.) every 24 hours for 14 consecutive days. All mice were sacrificed on day-15, 24 hours after the last rMETase administration. rMETase effectively reduced tumor growth compared to untreated control. The methionine level both of plasma and supernatants derived from sonicated tumors was lower in the rMETase group. Body weight did not significantly differ at any time points between the 2 groups. The present study is the first demonstrating rMETase efficacy in a PDOX model, suggesting potential clinical development, especially in recalcitrant cancers such as Ewing's sarcoma

Behavioral Health in Rural America: Understanding Citizen Perceptions and Willingness to Respond to Community Needs

Amid nationwide efforts to address behavioral health needs, rural communities often face unique challenges and a lack of resources. This study presents a bottom-up approach used by one rural community in the Midwest to respond to their needs regarding mental health and substance use. A survey instrument was developed from interviews with community stakeholders and disseminated in both online and paper formats. The survey sought to understand citizen perspectives regarding quality of life, barriers to treatment, and willingness to engage in efforts to address the community’s needs. Data from 1,303 respondents (71.5% women, 54.7% income \u3c$42,000) were analyzed using descriptive statistics and chi-square analyses. Results indicate that cost of treatment, shame, and lack of privacy were a barrier for most citizens’ treatment-seeking behavior. In addition, many citizens were willing to engage in strategies to address the community’s needs, including increased county spending, forming a neighborhood watch, and donating money. Differences associated with gender and income emerged across perceptions and willingness to support efforts. Implications for community efforts are discussed

25-Hydroxyvitamin D Depletion Does Not Exacerbate MPTP-Induced Dopamine Neuron Damage in Mice

Recent clinical evidence supports a link between 25-hydroxyvitamin D insufficiency (serum 25-hydroxyvitamin D [25(OH)D] levels <30 ng/mL) and Parkinson’s disease. To investigate the effect of 25(OH)D depletion on neuronal susceptibility to toxic insult, we induced a state of 25(OH)D deficiency in mice and then challenged them with the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). We found there was no significant difference between control and 25(OH)D-deficient animals in striatal dopamine levels or dopamine transporter and tyrosine hydroxylase expression after lesioning with MPTP. Additionally, we found no difference in tyrosine hydroxylase expression in the substantia nigra pars compacta. Our data suggest that reducing 25(OH)D serum levels in mice has no effect on the vulnerability of nigral dopaminergic neurons in vivo in this model system of parkinsonism

Centre selection for clinical trials and the generalisability of results: a mixed methods study.

BACKGROUND: The rationale for centre selection in randomised controlled trials (RCTs) is often unclear but may have important implications for the generalisability of trial results. The aims of this study were to evaluate the factors which currently influence centre selection in RCTs and consider how generalisability considerations inform current and optimal practice. METHODS AND FINDINGS: Mixed methods approach consisting of a systematic review and meta-summary of centre selection criteria reported in RCT protocols funded by the UK National Institute of Health Research (NIHR) initiated between January 2005-January 2012; and an online survey on the topic of current and optimal centre selection, distributed to professionals in the 48 UK Clinical Trials Units and 10 NIHR Research Design Services. The survey design was informed by the systematic review and by two focus groups conducted with trialists at the Birmingham Centre for Clinical Trials. 129 trial protocols were included in the systematic review, with a total target sample size in excess of 317,000 participants. The meta-summary identified 53 unique centre selection criteria. 78 protocols (60%) provided at least one criterion for centre selection, but only 31 (24%) protocols explicitly acknowledged generalisability. This is consistent with the survey findings (n = 70), where less than a third of participants reported generalisability as a key driver of centre selection in current practice. This contrasts with trialists' views on optimal practice, where generalisability in terms of clinical practice, population characteristics and economic results were prime considerations for 60% (n = 42), 57% (n = 40) and 46% (n = 32) of respondents, respectively. CONCLUSIONS: Centres are rarely enrolled in RCTs with an explicit view to external validity, although trialists acknowledge that incorporating generalisability in centre selection should ideally be more prominent. There is a need to operationalize 'generalisability' and incorporate it at the design stage of RCTs so that results are readily transferable to 'real world' practice