Antigenized antibodies expressing Vβ8.2 TCR peptides immunize against rat experimental allergic encephalomyelitis

BACKGROUND: Immunity against the T cell receptor (TCR) is considered to play a central role in the regulation of experimental allergic encephalomyelitis (EAE), a model system of autoimmune disease characterized by a restricted usage of TCR genes. Methods of specific vaccination against the TCR of pathogenetic T cells have included attenuated T cells and synthetic peptides from the sequence of the TCR. These approaches have led to the concept that anti-idiotypic immunity against antigenic sites of the TCR, which are a key regulatory element in this disease. METHODS: The present study in the Lewis rat used a conventional idiotypic immunization based on antigenized antibodies expressing selected peptide sequences of the Vβ8.2 TCR ((93)ASSDSSNTE(101 )and (39)DMGHGLRLIHYSYDVNSTEKG(59)). RESULTS: The study demonstrates that vaccination with antigenized antibodies markedly attenuates, and in some instances, prevents clinical EAE induced with the encephalitogenic peptide (68)GSLPQKSQRSQDENPVVHF(88 )in complete Freunds' adjuvant (CFA). Antigenized antibodies induced an anti-idiotypic response against the Vβ8.2 TCR, which was detected by ELISA and flowcytometry. No evidence was obtained of a T cell response against the corresponding Vβ8.2 TCR peptides. CONCLUSIONS: The results indicate that antigenized antibodies expressing conformationally-constrained TCR peptides are a simple means to induce humoral anti-idiotypic immunity against the TCR and to vaccinate against EAE. The study also suggests the possibility to target idiotypic determinants of TCR borne on pathogenetic T cells to vaccinate against disease

Tumor Stress Inside Out: Cell-Extrinsic Effects of the Unfolded Protein Response in Tumor Cells Modulate the Immunological Landscape of the Tumor Microenvironment

The unfolded protein response (UPR) is a eukaryotic cellular adaptive mechanism that functions to cope with stress of the endoplasmic reticulum (ER). Accumulating evidence demonstrates that the tumor microenvironment contains stressors that elicit a UPR, which has been demonstrated to be a cell-intrinsic mechanism crucial for tumorigenesis. In addition, the UPR is a source of proinflammatory signaling whose downstream mediators may hamper antitumor immunity. We discuss how the UPR may impair Ag presentation, which could result in defective T cell priming, also leading to tumor escape and growth. Further, we discuss the recent finding that ER stress and attendant proinflammation can be transmitted from ER-stressed tumor cells to myeloid cells. The ideas presented suggest that, in addition to being a cell-intrinsic mechanism of tumor survival, the tumor UPR can serve as a cell-extrinsic regulator of tumorigenesis by remodeling the immune response in the tumor microenvironment

Elevated neoantigen levels in tumors with somatic mutations in the HLA-A, HLA-B, HLA-C and B2M genes.

BackgroundThe major histocompatibility complex class I (MHC-I) molecule is a protein complex that displays intracellular peptides to T cells, allowing the immune system to recognize and destroy infected or cancerous cells. MHC-I is composed of a highly polymorphic HLA-encoded alpha chain that binds the peptide and a Beta-2-microglobulin (B2M) protein that acts as a stabilizing scaffold. HLA mutations have been implicated as a mechanism of immune evasion during tumorigenesis, and B2M is considered a tumor suppressor gene. However, the implications of somatic HLA and B2M mutations have not been fully explored in the context of antigen presentation via the MHC-I molecule during tumor development. To understand the effect that B2M and HLA MHC-I molecule mutations have on mutagenesis, we analyzed the accumulation of mutations in patients from The Cancer Genome Atlas according to their MHC-I molecule mutation status.ResultsSomatic B2M and HLA mutations in microsatellite stable tumors were associated with higher overall mutation burden and a larger fraction of HLA-binding neoantigens when compared to B2M and HLA wild type tumors. B2M and HLA mutations were highly enriched in patients with microsatellite instability. B2M mutations tended to occur relatively early during patients' respective tumor development, whereas HLA mutations were either early or late events. In addition, B2M and HLA mutated patients had higher levels of immune infiltration by natural killer and CD8+ T cells and higher levels of cytotoxicity.ConclusionsOur findings add to a growing body of evidence that somatic B2M and HLA mutations are a mechanism of immune evasion by demonstrating that such mutations are associated with a higher load of neoantigens that should be presented via MHC-I

Extracellular vesicles produced in B cells deliver tumor suppressor miR-335 to breast cancer cells disrupting oncogenic programming in vitro and in vivo.

The successful implementation of miRNA (miR) therapies in humans will ultimately rely on the use of vehicles with improved cellular delivery capability. Here we tested a new system that leverages extracellular vesicles (EVs) laden with a tumor suppressor miRNA (miR-335) produced in B cells by plasmid DNA induction (iEVs). We demonstrate that iEVs-335 efficiently and durably restored the endogenous miR-335 pool in human triple negative breast cancer cells, downregulated the expression of the miR-335 target gene SOX4 transcription factor, and markedly inhibited tumor growth in vivo. Remarkably, iEVs-335 mediated transcriptional effects that persisted in tumors after 60 days post orthotopic implantation. Genome-wide RNASeq analysis of cancer cells treated in vitro with iEVs-335 showed the regulation of a discrete number of genes only, without broad transcriptome perturbations. This new technology may be ideally suited for therapies aimed to restore tumor suppressor miRNAs in cancer cells, disrupting the oncogenic program established after escape from miRNA control

Petrology and geochemistry of the back-arc lithospheric mantle beneath eastern Payunia (La Pampa, Argentina): evidence from Agua Poca peridotite xenoliths

This paper presents the results of new petrochemical studies carried out on mantle xenoliths hosted in Pleistocene basaltic rocks from the Agua Poca volcano in central-western Argentina. Mantle xenoliths studied are shown to be mainly anhydrous spinel lherzolites with minor amounts of harzburgite and banded pyroxenite, showing highly variable equilibrium temperatures ranging from 820°C to 1030°C at 1.0 to 2.0 GPa. This constitutes evidence that the mantle xenoliths are representative of a large portion of the lithospheric mantle column and that the geothermal gradient is not very elevated as reported in some other Patagonian provinces. Geochemical characteristics of clinopyroxene in the mantle xenoliths allow classification into two groups; Groups 1 and 2. Group 1 contains most of the lherzolites and has light-REE depletion, with slightly positive anomalies of Eu in some samples and extreme Nb and Ta depletion. Group 2 consists of two harzburgitic samples, has flat REE patterns with lower Sm to Lu concentrations, with enriched Sr and negative HFSE anomalies. Based on mineral and residua compositions estimated assuming equilibrium with clinopyroxenes, Group 1 can be considered to be refractory residua after up to 7%, non-modal, near-fractional melting of a spinel-facies Primitive Mantle. Group 2 can be considered to be after ca. 13% of partial melting. It is inferred that partial melting events in the lithospheric mantle beneath the Agua Poca occurred in different ages since the Proterozoic, but compared with Group 1, the metasomatic overprint is dominant in Group 2 mantle xenoliths. The calculated melt compositions from Group 2 are interpreted to be transient liquid compositions developed during melt-peridotite interaction, and are different from the host alkaline basalts. The HFSE-depleted composition estimated for the rising melt suggests the presence of a slab-derived component, although the possibility cannot be disregarded (on the basis of present data) that such a geochemical feature is due to segregation of HFSE-bearing minerals during the interaction with the peridotite. Thus, we attribute the metasomatic agent to a basaltic melt and to a minor amount of slab-derived fluids.Fil: Bertotto, Gustavo Walter. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; ArgentinaFil: Mazzucchelli, Maurizio. Università di Modena e Reggio Emilia. Dipartimento di Scienze Chimiche e Geologiche; ItaliaFil: Zanetti, Alberto. Università di Pavia. Istituto di Geoscienze e Georisorse; Italia. Consiglio Nazionale delle Ricerche; ItaliaFil: Vannucci, Riccardo. Università di Pavia. Dipartamento di Scienze della Terra; Itali

Petrology and geochemistry of the back-arc lithospheric mantle beneath eastern Payunia (La Pampa, Argentina): evidence from Agua Poca peridotite xenoliths

This paper presents the results of new petrochemical studies carried out on mantle xenoliths hosted in Pleistocene basaltic rocks from the Agua Poca volcano in central-western Argentina. Mantle xenoliths studied are shown to be mainly anhydrous spinel lherzolites with minor amounts of harzburgite and banded pyroxenite, showing highly variable equilibrium temperatures ranging from 820°C to 1030°C at 1.0 to 2.0 GPa. This constitutes evidence that the mantle xenoliths are representative of a large portion of the lithospheric mantle column and that the geothermal gradient is not very elevated as reported in some other Patagonian provinces. Geochemical characteristics of clinopyroxene in the mantle xenoliths allow classification into two groups; Groups 1 and 2. Group 1 contains most of the lherzolites and has light-REE depletion, with slightly positive anomalies of Eu in some samples and extreme Nb and Ta depletion. Group 2 consists of two harzburgitic samples, has flat REE patterns with lower Sm to Lu concentrations, with enriched Sr and negative HFSE anomalies. Based on mineral and residua compositions estimated assuming equilibrium with clinopyroxenes, Group 1 can be considered to be refractory residua after up to 7%, non-modal, near-fractional melting of a spinel-facies Primitive Mantle. Group 2 can be considered to be after ca. 13% of partial melting. It is inferred that partial melting events in the lithospheric mantle beneath the Agua Poca occurred in different ages since the Proterozoic, but compared with Group 1, the metasomatic overprint is dominant in Group 2 mantle xenoliths. The calculated melt compositions from Group 2 are interpreted to be transient liquid compositions developed during melt-peridotite interaction, and are different from the host alkaline basalts. The HFSE-depleted composition estimated for the rising melt suggests the presence of a slab-derived component, although the possibility cannot be disregarded (on the basis of present data) that such a geochemical feature is due to segregation of HFSE-bearing minerals during the interaction with the peridotite. Thus, we attribute the metasomatic agent to a basaltic melt and to a minor amount of slab-derived fluids.Fil: Bertotto, Gustavo Walter. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; ArgentinaFil: Mazzucchelli, Maurizio. Università di Modena e Reggio Emilia. Dipartimento di Scienze Chimiche e Geologiche; ItaliaFil: Zanetti, Alberto. Università di Pavia. Istituto di Geoscienze e Georisorse; Italia. Consiglio Nazionale delle Ricerche; ItaliaFil: Vannucci, Riccardo. Università di Pavia. Dipartamento di Scienze della Terra; Itali

Geochemistry of sapphirine-apatite-calcite-bearing gabbroic dykes from the Finero Phlogopite Peridotite (Ivrea-Verbano Zone): evidence for multistage interaction with the ambient peridotite

The Finero Phlogopite-Peridotite (FPP) is a mantle unit outcropping in the northernmost tip of the Ivrea-Verbano Zone (IVZ, Southern Alps). It shows a virtually complete recrystallization due to pervasive to channelled melt migration. The pervasive metasomatism formed a main lithologic association constituted by phlogopite harzburgites associated to phlogopite pyroxenites (mainly olivine-websterites, websterites and orthopyroxenites). These lithologies are also rich in amphibole and do not show significant chemical gradients among them (Zanetti et al., 1999). The channelled migration stages formed dunite bodies, which sometimes contain stratiform chromitites and, more rarely, pyroxenite layers similar to those associated to phlogopite harzburgite. The FPP also shows a discrete number of other, subordinate rock-types, which are characterised by the presence of apatite usually associated to carbonates (i.e. calcite or dolomite) and exhibit marked modal and chemical gradients with respect to the host phlogopite harzburgite. Examples of these lithologies are apatite-dolomitebearing wehrlites and harzburgites (e.g. Zanetti et al. 1999; Morishita et al., 2008), apatite-calcite zircon-syenites and hornblendites. Ar-Ar amphibole analysis and U-Pb zircon and apatite data return Triassic ages for these rocks, which have been considered to document the time of melt/fluid injection. Notwithstanding the apparent mineralogical and chemical differences with the main lithologic sequences, apatite-carbonates-bearing rocks have been frequently interpreted as cogenetic to phlogopite harzburgites. To debate the petrogenesis of these rocks, a detailed field, petrological and geochemical investigation has been carried out on a swarm of apatite-calcite-bearing gabbroic veins that randomly cut the main lithologic association. Preliminary investigation evidenced as these veins show complex metasomatic haloes and a symmetric internal layering, characterised by crystallisation of magmatic sapphirine (Giovanardi et al., 2013). The mineral assemblage of the veins is dominated by titanian pargasite towards the host peridotite and by plagioclase at the vein centre. The veins also present phlogopite and spinel. Field and petrographic evidence, major and trace element data and the O isotopic composition of such gabbroic veins indicate that they formed at shallow mantle conditions by multistage fractional crystallisation of a migrating melt unrelated to those forming phlogopite harzburgites. Besides, local strong enrichments in LILE, LREE and 18O in vein minerals confirm that such melt was deeply modified by interaction with the host phlogopite peridotite. The genetic relationships with other intrusive events recorded by the FPP and the associated crustal sequence will be addressed with the aim of placing new constraints on the petrologic and geodynamic evolution of the IVZ

Melt-Rock Interaction at Mantle Conditions: Evidences from Finero Gabbroic Dykes

The Finero Phlogopite-Peridotite (FPP) is a worldwide famous mantle massif recrystallized through several events of melt migrations. These events have enriched the FPP in hydrous phases and crustal components and have been commonly interpreted as related to a subduction/post orogenic geodynamic setting. The last of these metasomatic events has produced composite sapphirine-bearing gabbroic dykes, interpreted as the result of the interaction of channelized migrating melts with the host rock in a two-steps intrusion process. In the first step, the melt reacted with the FPP rocks and evolved by fractional crystallization of amphibole cumulates. In the second step, the evolved melt reacted with the first cumulates producing magmatic sapphirine and segregating plagioclase-rich bands containing abundant apatites at the nucleus of the dike. New data suggest, however, a more complex evolution. New O and in situ Sr isotopes on minerals suggest that the gabbroic dykes have evolved from melt(s) that progressively were contaminated by the interaction with the FPP rocks during its fractionation. The δ18O increases from 5.81‰ in orthopyroxenes at the dykes border to ~6.90‰ in cumulitic amphiboles and 8.60‰ in plagioclases. The 87Sr/86Sr values for plagioclase and coexisting apatite show isotopic disequilibrium between the two phases (plagioclases at 0.70474 ± 0.00033, n=23, and apatites at 0.70369 ± 0.00025, n=6). These isotopic variations could be explained with an AFC-like process between mantle-derived melt(s) and a crustal-enriched host (the FPP). In situ Sr isotope analyses were performed at the CIGS laboratory of the Università di Modena e Reggio Emilia using a Thermo Fisher Scientific Neptune™ coupled to a 213 nm Nd:YAG laser ablation system (New Wave Research™). During the analytical sessions a new in house plagioclase reference material for Rb-Sr systematic, named BC84, has been successfully tested and used

Prostate cancer cells undergoing ER stress in vitro and in vivo activate transcription of pro-inflammatory cytokines

Navin R Mahadevan, Antonio Fernandez, Jeffrey J Rodvold, Gonzalo Almanza, Maurizio ZanettiThe Laboratory of Immunology, Department of Medicine and Moores Cancer Center, University of California, San Diego, USABackground: Several micro-environmental and cell-intrinsic stimuli cause tumor cells to undergo endoplasmic reticulum (ER) stress in vivo. The occurrence of an ER stress response has been associated with tumor progression and angiogenesis. Recently, we found that pharmacological induction of ER stress in B lymphoma cells upregulates the transcription of several pro-inflammatory cytokines.Results: Here, we show that transgenic adenocarcinoma of the mouse prostate (TRAMP) C1 murine prostate cancer cells induced to undergo ER stress in vitro activate the transcription of interleukin 6 (IL-6), interleukin 23p19 (IL-23p19), and tumor necrosis factor a (TNF-a). Furthermore we show that TRAMP C1 tumors growing in vivo spontaneously experience ER stress and that transcription of IL-6, IL-23p19, and TNF-a correlates with the in vivo ER stress response.Conclusions: These results suggest that an ER stress response in prostate cancer cells activates a program of pro-inflammatory cytokine transcription. A possible implication of this finding is that cancer cells may use the ER stress response to modify their microenvironment.Keywords: unfolded protein response, tumorigenesis, inflammatio

Mantle-Derived Corundum-Bearing Felsic Dykes May Survive Only within the Lower (Refractory/Inert) Crust: Evidence from Zircon Geochemistry and Geochronology (Ivrea–Verbano Zone, Southern Alps, Italy)

Corundum-rich (up to 55 vol.%) felsic dykes formed with albite, +/- K-feldspar, +/- hercynite and +/- biotite-siderophyllite cut the lower crustal rocks exposed in the Ivrea–Verbano Zone (NW Italy). Zircon is an abundant accessory mineral and its investigation through laser ablation-inductively coupled plasma (multi-collector)-mass spectrometer (LA-ICP-(MC)MS) has allowed results to directly constrain the timing of emplacement, as well as petrology and geochemistry of parental melts. Zircons are characterized by very large concentration in rare earth elements (REE), Th, U, Nb and Ta, and negative Eu anomaly. U–Pb analysis points to Norian emplacement ages (223 +/- 7 Ma and 224 +/- 6 Ma), whereas large positive EHf(t) values (+13 on average) indicate a derivation from depleted to mildly enriched mantle source. The mantle signature and the corundum oversaturation were preserved thanks to limited crustal contamination of the host, high-temperature refractory granulites and mafic intrusives. According to the geochemical data and to the evidence of the development of violent explosions into the conduits, it is proposed that dykes segregated from peraluminous melts produced by exsolution processes affecting volatile-rich differentiates during alkaline magmatism. This work provides robust evidence about the transition of the geochemical affinity of Southern Alps magmatism from orogenic-like to anorogenic during Norian time, linked to a regional uprising of the asthenosphere and change of tectonic regime