Cancer Immunotherapy Targeting T Cell Costimulatory Molecules

According to the American Cancer Society, over 1.5 million new cancer cases will be diagnosed this year, a figure that is expected to rise with the aging population. Chemotherapy and radiation are the current "gold standards" for cancer treatment, but these therapies are marginally effective, toxic, and serve to diminish the quality of life for cancer patients. Immunotherapy represents an attractive alternative to these traditional treatment regimens. Despite overwhelming evidence that the immune system is capable of recognizing and eliminating tumors, both spontaneously and in response to immune-based therapy, such protection is abrogated in the face of compensatory immunosuppressive events characteristic of progressive disease. Thus, a major goal of novel immune-based therapies is the coordinate silencing of regulatory circuits and amplification of protective T cell function.While immune modulating reagents that trigger the T cell costimulatory molecules OX40 and GITR are currently being evaluated in early-phase clinical trials, little pre-clinical information is available regarding the efficacy and mechanism(s) of action for these agents in the setting of advanced, well-established disease. To further characterize the molecular, cellular, and treatment-associated consequences of OX40 and GITR engagement, novel agonistic reagents directed against murine OX40 and GITR (ligand-Fc fusion proteins) were recently constructed and characterized in vitro. We now show that the growth of well-established, day 17 sarcomas is significantly inhibited or ablated by a short course of either treatment, with OX40L-Fc demonstrating superior anti-tumor efficacy over GITRL-Fc at comparable dosing. Both treatments were capable of eliminating regulatory T cells within tumors, inducing profound proliferation of T effector cells in the tumor-draining lymph node, and promoting the recruitment of these expanded effector cells to the tumor microenvironment. However, OX40L-Fc therapy mediated additional, T cell-independent effects, including the activation of tumor-localized dendritic and endothelial cell subsets. These changes rendered the tumor microenvironment more immunogenic and permissive to the infiltration of treatment-induced, protective immune cells. The pleiotropic anti-tumor effects demonstrated in this model by OX40L-Fc, and to a lesser extent GITRL-Fc, strongly supports the further translation of such modalities into human clinical trials, either as single agents or in the context of combinational immunotherapy

Short-time inertial response of viscoelastic fluids measured with Brownian motion and with active probes

We have directly observed short-time stress propagation in viscoelastic fluids using two optically trapped particles and a fast interferometric particle-tracking technique. We have done this both by recording correlations in the thermal motion of the particles and by measuring the response of one particle to the actively oscillated second particle. Both methods detect the vortex-like flow patterns associated with stress propagation in fluids. This inertial vortex flow propagates diffusively for simple liquids, while for viscoelastic solutions the pattern spreads super-diffusively, dependent on the shear modulus of the medium

Shape Changes of Self-Assembled Actin Bilayer Composite Membranes

We report the self-assembly of thin actin shells beneath the membranes of giant vesicles. Ion-carrier mediated influx of Mg2+ induces actin polymerization in the initially spherical vesicles. Buckling of the vesicles and the formation of blisters after thermally induced bilayer expansion is demonstrated. Bilayer flickering is dominated by tension generated by its coupling to the actin cortex. Quantitative flicker analysis suggests the bilayer and the actin cortex are separated by 0.4 \mum to 0.5 \mum due to undulation forces.Comment: pdf-file, has been accepted by PR

Performance of commercial corn hybrids in Illinois, 1961-1963

Chiefly tables.Cover title

Two fingerprinting sets for Humulus lupulus based on KASP and microsatellite markers

Verification of clonal identity of hop (Humulus lupulus L.) cultivars within breeding programs and germplasm collections is vital to conserving genetic resources. Accurate and economic DNA-based tools are needed in dioecious hop to confirm identity and parentage, neither of which can be reliably determined from morphological observations. In this study, we developed two fingerprinting sets for hop: a 9-SSR fingerprinting set containing high-core repeats that can be run in a single PCR reaction and a kompetitive allele specific PCR (KASP) assay of 25 single nucleotide polymorphisms (SNPs). The SSR set contains a sex-linked primer pair, HI-AGA7, that was used to genotype 629 hop accessions from the US Department of Agriculture (USDA) National Clonal Germplasm Repository (NCGR), the USDA Forage Seed and Cereal Research (FSCR), and the University of Nebraska-Lincoln (UNL) collections. The SSR set identified unique genotypes except for 89 sets of synonymous samples. These synonyms included: cultivars with different designations, the same cultivars from different sources, heat-treated clones, and clonal variants. Population structure analysis clustered accessions into wild North American (WNA) and cultivated groups. Diversity was slightly higher in the cultivated samples due to larger sample size. Parentage and sib-ship analyses were used to identify true-to-type cultivars. The HI-AGA7 marker generated two male- and nine female-specific alleles among the cultivated and WNA samples. The SSR and KASP fingerprinting sets were compared in 190 samples consisting of cultivated and WNA accession for their ability to confirm identity and assess diversity and population structure. The SSR fingerprinting set distinguished cultivars, selections and WNA accessions while the KASP assays were unable to distinguish the WNA samples and had lower diversity estimates than the SSR set. Both fingerprinting sets are valuable tools for identity confirmation and parentage analysis in hop for different purposes. The 9-SSR assay is cost efficient when genotyping a small number of wild and cultivated hop samples (\u3c96) while the KASP assay is easy to interpret and cost efficient for genotyping a large number of cultivated samples (multiples of 96)

Achieving Optimal Growth through Product Feedback Inhibition in Metabolism

Recent evidence suggests that the metabolism of some organisms, such as Escherichia coli, is remarkably efficient, producing close to the maximum amount of biomass per unit of nutrient consumed. This observation raises the question of what regulatory mechanisms enable such efficiency. Here, we propose that simple product-feedback inhibition by itself is capable of leading to such optimality. We analyze several representative metabolic modules—starting from a linear pathway and advancing to a bidirectional pathway and metabolic cycle, and finally to integration of two different nutrient inputs. In each case, our mathematical analysis shows that product-feedback inhibition is not only homeostatic but also, with appropriate feedback connections, can minimize futile cycling and optimize fluxes. However, the effectiveness of simple product-feedback inhibition comes at the cost of high levels of some metabolite pools, potentially associated with toxicity and osmotic imbalance. These large metabolite pool sizes can be restricted if feedback inhibition is ultrasensitive. Indeed, the multi-layer regulation of metabolism by control of enzyme expression, enzyme covalent modification, and allostery is expected to result in such ultrasensitive feedbacks. To experimentally test whether the qualitative predictions from our analysis of feedback inhibition apply to metabolic modules beyond linear pathways, we examine the case of nitrogen assimilation in E. coli, which involves both nutrient integration and a metabolic cycle. We find that the feedback regulation scheme suggested by our mathematical analysis closely aligns with the actual regulation of the network and is sufficient to explain much of the dynamical behavior of relevant metabolite pool sizes in nutrient-switching experiments