Detection and Characterizationof Cellular Immune Responses Using Peptide–MHC Microarrays

The detection and characterization of antigen-specific T cell populations is critical for understanding the development and physiology of the immune system and its responses in health and disease. We have developed and tested a method that uses arrays of peptide–MHC complexes for the rapid identification, isolation, activation, and characterization of multiple antigen-specific populations of T cells. CD4(+) or CD8(+) lymphocytes can be captured in accordance with their ligand specificity using an array of peptide–MHC complexes printed on a film-coated glass surface. We have characterized the specificity and sensitivity of a peptide–MHC array using labeled lymphocytes from T cell receptor transgenic mice. In addition, we were able to use the array to detect a rare population of antigen-specific T cells following vaccination of a normal mouse. This approach should be useful for epitope discovery, as well as for characterization and analysis of multiple epitope-specific T cell populations during immune responses associated with viral and bacterial infection, cancer, autoimmunity, and vaccination

Marked Differences in Human Melanoma Antigen-Specific T Cell Responsiveness after Vaccination Using a Functional Microarray

BACKGROUND: In contrast to many animal model studies, immunotherapeutic trials in humans suffering from cancer invariably result in a broad range of outcomes, from long-lasting remissions to no discernable effect. METHODS AND FINDINGS: In order to study the T cell responses in patients undergoing a melanoma-associated peptide vaccine trial, we have developed a high-throughput method using arrays of peptide-major histocompatibility complexes (pMHC) together with antibodies against secreted factors. T cells were specifically immobilized and activated by binding to particular pMHCs. The antibodies, spotted together with the pMHC, specifically capture cytokines secreted by the T cells. This technique allows rapid, simultaneous isolation and multiparametric functional characterization of antigen-specific T cells present in clinical samples. Analysis of CD8+ lymphocytes from ten melanoma patients after peptide vaccination revealed a diverse set of patient- and antigen-specific profiles of cytokine secretion, indicating surprising differences in their responsiveness. Four out of four patients who showed moderate or greater secretion of both interferon-γ (IFNγ) and tumor necrosis factor-α (TNFα) in response to a gp100 antigen remained free of melanoma recurrence, whereas only two of six patients who showed discordant secretion of IFNγ and TNFα did so. CONCLUSION: Such multiparametric analysis of T cell antigen specificity and function provides a valuable tool with which to dissect the molecular underpinnings of immune responsiveness and how this information correlates with clinical outcome

Partners No More: Relational Transformation and the Turn to Litigation in Two Conservationist Organizations

The rise in litigation against administrative bodies by environmental and other political interest groups worldwide has been explained predominantly through the liberalization of standing doctrines. Under this explanation, termed here the floodgate model, restrictive standing rules have dammed the flow of suits that groups were otherwise ready and eager to pursue. I examine this hypothesis by analyzing processes of institutional transformation in two conservationist organizations: the Sierra Club in the United States and the Society for the Protection of Nature in Israel (SPNI). Rather than an eagerness to embrace newly available litigation opportunities, as the floodgate model would predict, the groups\u27 history reveals a gradual process of transformation marked by internal, largely intergenerational divisions between those who abhorred conflict with state institutions and those who saw such conflict as not only appropriate but necessary to the mission of the group. Furthermore, in contrast to the pluralist interactions that the floodgate model imagines, both groups\u27 relations with pertinent agencies in earlier eras better accorded with the partnership-based corporatist paradigm. Sociolegal research has long indicated the importance of relational distance to the transformation of interpersonal disputes. I argue that, at the group level as well, the presence or absence of a (national) partnership-centered relationship determines propensities to bring political issues to court. As such, well beyond change in groups\u27 legal capacity and resources, current increases in levels of political litigation suggest more fundamental transformations in the structure and meaning of relations between citizen groups and the state

A novel Alzheimer disease locus located near the gene encoding tau protein

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this recordAPOE ε4, the most significant genetic risk factor for Alzheimer disease (AD), may mask effects of other loci. We re-analyzed genome-wide association study (GWAS) data from the International Genomics of Alzheimer's Project (IGAP) Consortium in APOE ε4+ (10 352 cases and 9207 controls) and APOE ε4- (7184 cases and 26 968 controls) subgroups as well as in the total sample testing for interaction between a single-nucleotide polymorphism (SNP) and APOE ε4 status. Suggestive associations (P<1 × 10-4) in stage 1 were evaluated in an independent sample (stage 2) containing 4203 subjects (APOE ε4+: 1250 cases and 536 controls; APOE ε4-: 718 cases and 1699 controls). Among APOE ε4- subjects, novel genome-wide significant (GWS) association was observed with 17 SNPs (all between KANSL1 and LRRC37A on chromosome 17 near MAPT) in a meta-analysis of the stage 1 and stage 2 data sets (best SNP, rs2732703, P=5·8 × 10-9). Conditional analysis revealed that rs2732703 accounted for association signals in the entire 100-kilobase region that includes MAPT. Except for previously identified AD loci showing stronger association in APOE ε4+ subjects (CR1 and CLU) or APOE ε4- subjects (MS4A6A/MS4A4A/MS4A6E), no other SNPs were significantly associated with AD in a specific APOE genotype subgroup. In addition, the finding in the stage 1 sample that AD risk is significantly influenced by the interaction of APOE with rs1595014 in TMEM106B (P=1·6 × 10-7) is noteworthy, because TMEM106B variants have previously been associated with risk of frontotemporal dementia. Expression quantitative trait locus analysis revealed that rs113986870, one of the GWS SNPs near rs2732703, is significantly associated with four KANSL1 probes that target transcription of the first translated exon and an untranslated exon in hippocampus (P≤1.3 × 10-8), frontal cortex (P≤1.3 × 10-9) and temporal cortex (P≤1.2 × 10-11). Rs113986870 is also strongly associated with a MAPT probe that targets transcription of alternatively spliced exon 3 in frontal cortex (P=9.2 × 10-6) and temporal cortex (P=2.6 × 10-6). Our APOE-stratified GWAS is the first to show GWS association for AD with SNPs in the chromosome 17q21.31 region. Replication of this finding in independent samples is needed to verify that SNPs in this region have significantly stronger effects on AD risk in persons lacking APOE ε4 compared with persons carrying this allele, and if this is found to hold, further examination of this region and studies aimed at deciphering the mechanism(s) are warranted

Nucleation and growth kinetics of RaxBa1-xSO4 solid solution in NaCl aqueous solutions

Co-precipitation of Ra and Ba in barite (i.e., the formation of a RaxBa1-xSO4 solid solution) has long been established as an important process that has the potential to control Ra concentration. This process is commonly described by a distribution model. Ample studies have shown that the key parameter of this model, the partition coefficient, varies in the range of 1–2 as a function of temperature, salinity and precipitation kinetics of the RaxBa1-xSO4 solid solution. This roughly twofold change in the partition coefficient may lead to large differences in the concentration of dissolved Ra. The present study systematically investigated the co-precipitation kinetics of the RaxBa1-xSO4 solid solution from aqueous solutions up to 5.9 mol kg-1 H2O NaCl, circum-neutral pH and at ambient temperature. Laboratory batch experiments designed to follow the nucleation of the RaxBa1-xSO4 solid solution and the co-precipitation kinetics of Ba and Ra from aqueous solutions which were initially supersaturated with respect to barite (degree of supersaturation, ßbarite = 20 ± 2). The following empirical law describes the dependence of the activity-based partition coefficient, K¨D,barite, on the degree of supersaturation, ßbarite: = 20 ± 2). K¨D,barite =(199 ± 0,05) - (0,58 ± 0,06)log(ßbarite). This empirical law is in good agreement with other literature data. The outcomes of the empirical law are compared to the prediction of a model for the nucleation of two-dimensional islands

Evolution of electrical resistivity and NMR during early-stage maturation of an organic-rich chalk

This paper studies the changes in the electrical resistivity of an organic-rich source rock chalk containing a Type-IIs kerogen during early-stage maturation of the kerogen. As bitumen is generated during maturation, the chalk changes from water wet to a complex mixed-wet pore system, with the micritic pore space remaining water wet while the bitumen expelled into the macro pores converts some pores to oil wet. This mixed-wet system results in non-Archie electrical behavior with pronounced curvature of the I-Sw curve. We find that this curvature is well fit by connectivity theory. The connectivity theory parameters are determined both on native state core and from artificial laboratory pyrolysis which generates bitumen. NMR is used to distinguish the brine in micritic, water-wet pore spaces from that in the intergranular, mixed-wet pore spaces, from which the observed connectivity parameters may be independently estimated. Comparison between electrical resistivity and dielectric logs show that the connectivity parameters match the laboratory results over a calcareous section more than 300 m thick

Gypsum Precipitation under Saline Conditions: Thermodynamics, Kinetics, Morphology, and Size Distribution

Gypsum (CaSO4·2H2O) is the most common sulfate mineral on Earth and is also found on Mars. It is an evaporitic mineral that predominantly precipitates from brines. In addition to its precipitation in natural environments, gypsum also forms an undesired scale in many industrial processes that utilize or produce brines. Thus, better insights into gypsum formation can contribute to the understanding of natural processes, as well as improving industrial practices. Subsequently, the thermodynamics, nucleation and crystal growth mechanisms and kinetics, and how these factors shape the morphology of gypsum have been widely studied. Over the last decade, the precipitation of gypsum under saline and hypersaline conditions has been the focus of several studies. However, to date, most of the thermodynamic data are derived from experiments with artificial solutions that have limited background electrolytes and have Ca2+/SO42− ratios that are similar to the 1:1 ratio in the mineral. Moreover, direct observations of the nucleation and growth processes of gypsum are still derived from experimental settings that can be described as having low ionic strength. Thus, the mechanisms of gypsum precipitation under conditions from which the mineral precipitates in many natural environments and industrial processes are still less well known. The present review focuses on the precipitation of gypsum from a range of aspects. Special attention is given to brines. The effects of ionic strength, brine composition, and temperature on the thermodynamic settings are broadly discussed. The mechanisms and rates of gypsum nucleation and growth, and the effect the thermodynamic properties of the brine have on these processes is demonstrated by recent microscopic and macroscopic observations. The morphology and size distribution of gypsum crystals precipitation is examined in the light of the precipitation processes that shape these properties. Finally, the present review highlights discrepancies between microscopic and macroscopic observations, and studies carried out under low and high ionic strengths. The special challenges posed by experiments with brines are also discussed. Thus, while this review covers contemporary literature, it also outlines further research that is required in order to improve our understanding of gypsum precipitation in natural environments and industrial settings

Gypsum Precipitation under Saline Conditions: Thermodynamics, Kinetics, Morphology, and Size Distribution

Gypsum (CaSO4·2H2O) is the most common sulfate mineral on Earth and is also found on Mars. It is an evaporitic mineral that predominantly precipitates from brines. In addition to its precipitation in natural environments, gypsum also forms an undesired scale in many industrial processes that utilize or produce brines. Thus, better insights into gypsum formation can contribute to the understanding of natural processes, as well as improving industrial practices. Subsequently, the thermodynamics, nucleation and crystal growth mechanisms and kinetics, and how these factors shape the morphology of gypsum have been widely studied. Over the last decade, the precipitation of gypsum under saline and hypersaline conditions has been the focus of several studies. However, to date, most of the thermodynamic data are derived from experiments with artificial solutions that have limited background electrolytes and have Ca2+/SO42− ratios that are similar to the 1:1 ratio in the mineral. Moreover, direct observations of the nucleation and growth processes of gypsum are still derived from experimental settings that can be described as having low ionic strength. Thus, the mechanisms of gypsum precipitation under conditions from which the mineral precipitates in many natural environments and industrial processes are still less well known. The present review focuses on the precipitation of gypsum from a range of aspects. Special attention is given to brines. The effects of ionic strength, brine composition, and temperature on the thermodynamic settings are broadly discussed. The mechanisms and rates of gypsum nucleation and growth, and the effect the thermodynamic properties of the brine have on these processes is demonstrated by recent microscopic and macroscopic observations. The morphology and size distribution of gypsum crystals precipitation is examined in the light of the precipitation processes that shape these properties. Finally, the present review highlights discrepancies between microscopic and macroscopic observations, and studies carried out under low and high ionic strengths. The special challenges posed by experiments with brines are also discussed. Thus, while this review covers contemporary literature, it also outlines further research that is required in order to improve our understanding of gypsum precipitation in natural environments and industrial settings