Activating neutrophils by co-administration of immunogenic recombinant modified vaccinia virus Ankara and granulocyte colony-stimulating factor for the treatment of malignant peripheral nerve sheath tumor.

Malignant peripheral nerve sheath tumor (MPNST) is a rare, aggressive soft-tissue sarcoma with a poor prognosis and is insensitive to immune checkpoint blockade (ICB) therapy. Loss-of-function of the histone modifying polycomb repressive complex 2 (PRC2) components, EED or SUZ12, is one of the main mechanisms of malignant transformation. In a murine model of MPNST, PRC2-loss tumors have an "immune desert" phenotype and intratumoral (IT) delivery immunogenic modified vaccinia virus Ankara (MVA) sensitized the PRC2-loss tumors to ICB. Here we show that IT MQ833, a second-generation recombinant modified vaccinia virus Ankara virus, results in neutrophil recruitment and activation and neutrophil-dependent tumor killing in the MPNST model. MQ833 was engineered by deleting three viral immune evasion genes, E5R, E3L, and WR199, and expressing three transgenes, including the two membrane-bound Flt3L and OX40L, and IL-12 with an extracellular matrix anchoring signal. Furthermore, we explored strategies to enhance anti-tumor effects of MQ833 by co-administration of granulocyte colony-stimulating factor (G-CSF).

Reprogramming tumor-associated macrophages to outcompete endovascular endothelial progenitor cells and suppress tumor neoangiogenesis.

Tumors develop by invoking a supportive environment characterized by aberrant angiogenesis and infiltration of tumor-associated macrophages (TAMs). In a transgenic model of breast cancer, we found that TAMs localized to the tumor parenchyma and were smaller than mammary tissue macrophages. TAMs had low activity of the metabolic regulator mammalian/mechanistic target of rapamycin complex 1 (mTORC1), and depletion of negative regulator of mTORC1 signaling, tuberous sclerosis complex 1 (TSC1), in TAMs inhibited tumor growth in a manner independent of adaptive lymphocytes. Whereas wild-type TAMs exhibited inflammatory and angiogenic gene expression profiles, TSC1-deficient TAMs had a pro-resolving phenotype. TSC1-deficient TAMs relocated to a perivascular niche, depleted protein C receptor (PROCR)-expressing endovascular endothelial progenitor cells, and rectified the hyperpermeable blood vasculature, causing tumor tissue hypoxia and cancer cell death. TSC1-deficient TAMs were metabolically active and effectively eliminated PROCR-expressing endothelial cells in cell competition experiments. Thus, TAMs exhibit a TSC1-dependent mTORC1-low state, and increasing mTORC1 signaling promotes a pro-resolving state that suppresses tumor growth, defining an innate immune tumor suppression pathway that may be exploited for cancer immunotherapy.

Reprogramming tumour-associated macrophages to outcompete cancer cells.

In metazoan organisms, cell competition acts as a quality control mechanism to eliminate unfit cells in favour of their more robust neighbours1,2. This mechanism has the potential to be maladapted, promoting the selection of aggressive cancer cells3-6. Tumours are metabolically active and are populated by stroma cells7,8, but how environmental factors affect cancer cell competition remains largely unknown. Here we show that tumour-associated macrophages (TAMs) can be dietarily or genetically reprogrammed to outcompete MYC-overexpressing cancer cells. In a mouse model of breast cancer, MYC overexpression resulted in an mTORC1-dependent 'winner' cancer cell state. A low-protein diet inhibited mTORC1 signalling in cancer cells and reduced tumour growth, owing unexpectedly to activation of the transcription factors TFEB and TFE3 and mTORC1 in TAMs. Diet-derived cytosolic amino acids are sensed by Rag GTPases through the GTPase-activating proteins GATOR1 and FLCN to control Rag GTPase effectors including TFEB and TFE39-14. Depletion of GATOR1 in TAMs suppressed the activation of TFEB, TFE3 and mTORC1 under the low-protein diet condition, causing accelerated tumour growth; conversely, depletion of FLCN or Rag GTPases in TAMs activated TFEB, TFE3 and mTORC1 under the normal protein diet condition, causing decelerated tumour growth. Furthermore, mTORC1 hyperactivation in TAMs and cancer cells and their competitive fitness were dependent on the endolysosomal engulfment regulator PIKfyve. Thus, noncanonical engulfment-mediated Rag GTPase-independent mTORC1 signalling in TAMs controls competition between TAMs and cancer cells, which defines a novel innate immune tumour suppression pathway that could be targeted for cancer therapy.

Assessing in vivo presentation of exogenous antigen in the tumor microenvironment in mice.

Here, we focus on tumor-associated macrophages (TAMs) in the PyMT model of breast cancer, detailing a protocol for assessing antigen presentation capabilities of immune populations of interest. We describe a stringent bone marrow chimera system to demonstrate presentation of exogenous antigen that is acquired and processed in the tumor microenvironment. We describe steps for testing antigen presentation activity of TAMs to CD8+ T cells in vivo and ex vivo and the requirement for the transcription factor IRF8 in this function. For complete details on the use and execution of this protocol, please refer to Nixon et al. (2022).1.

Control of tumor-associated macrophage responses by nutrient acquisition and metabolism.

Metazoan tissue specification is associated with integration of macrophage lineage cells in sub-tissular niches to promote tissue development and homeostasis. Oncogenic transformation, most prevalently of epithelial cell lineages, results in maladaptation of resident tissue macrophage differentiation pathways to generate parenchymal and interstitial tumor-associated macrophages that largely foster cancer progression. In addition to growth factors, nutrients that can be consumed, stored, recycled, or converted to signaling molecules have emerged as crucial regulators of macrophage responses in tumor. Here, we review how nutrient acquisition through plasma membrane transporters and engulfment pathways control tumor-associated macrophage differentiation and function. We also discuss how nutrient metabolism regulates tumor-associated macrophages and how these processes may be targeted for cancer therapy.

Tumor-associated macrophages expressing the transcription factor IRF8 promote T cell exhaustion in cancer.

Tumors are populated by antigen-presenting cells (APCs) including macrophage subsets with distinct origins and functions. Here, we examined how cancer impacts mononuclear phagocytic APCs in a murine model of breast cancer. Tumors induced the expansion of monocyte-derived tumor-associated macrophages (TAMs) and the activation of type 1 dendritic cells (DC1s), both of which expressed and required the transcription factor interferon regulatory factor-8 (IRF8). Although DC1s mediated cytotoxic T lymphocyte (CTL) priming in tumor-draining lymph nodes, TAMs promoted CTL exhaustion in the tumor, and IRF8 was required for TAMs' ability to present cancer cell antigens. TAM-specific IRF8 deletion prevented exhaustion of cancer-cell-reactive CTLs and suppressed tumor growth. Tumors from patients with immune-infiltrated renal cell carcinoma had abundant TAMs that expressed IRF8 and were enriched for an IRF8 gene expression signature. Furthermore, the TAM-IRF8 signature co-segregated with CTL exhaustion signatures across multiple cancer types. Thus, CTL exhaustion is promoted by TAMs via IRF8.

Nanoparticle-insertion scheme to decouple electron injection from laser evolution in laser wakefield acceleration.

A localized nanoparticle insertion scheme is developed to decouple electron injection from laser evolution in laser wakefield acceleration. Here we report the experimental realization of a controllable electron injection by the nanoparticle insertion method into a plasma medium, where the injection position is localized within the short range of 100 µm. Nanoparticles were generated by the laser ablation process of a copper blade target using a 3-ns 532-nm laser pulse with fluence above 100 J/cm2. The produced electron bunches with a beam charge above 300 pC and divergence of around 12 mrad show the injection probability over 90% after optimizing the ablation laser energy and the temporal delay between the ablation and the main laser pulses. Since this nanoparticle insertion method can avoid the disturbing effects of electron injection process on laser evolution, the stable high-charge injection method can provide a suitable electron injector for multi-GeV electron sources from low-density plasmas.

Improving the accuracy of hard photon emission by sigmoid sampling of the quantum-electrodynamic table in particle-in-cell Monte Carlo simulations.

Research on laser-plasma interaction in the quantum-electrodynamic (QED) regime has been greatly advanced by particle-in-cell and Monte Carlo simulations (PIC-MC). While these simulations are widely used, we find that a noticeable numerical error arises due to inappropriate implementation of the quantum process accounting for hard photon emission and pair production in the PIC-MC codes. The error stems from the low resolution of the QED table used to sample photon energy, which is generated in the logarithmic scale and cannot resolve high energy photons. We propose a sampling method via sigmoid function that handles both the low energy and high energy end of the photon emission spectrum. It guarantees the accuracy of PIC-MC algorithms for hard photon radiation and other related processes in the strong-field QED regime.

Mitocytosis, a migrasome-mediated mitochondrial quality-control process.

Damaged mitochondria need to be cleared to maintain the quality of the mitochondrial pool. Here, we report mitocytosis, a migrasome-mediated mitochondrial quality-control process. We found that, upon exposure to mild mitochondrial stresses, damaged mitochondria are transported into migrasomes and subsequently disposed of from migrating cells. Mechanistically, mitocytosis requires positioning of damaged mitochondria at the cell periphery, which occurs because damaged mitochondria avoid binding to inward motor proteins. Functionally, mitocytosis plays an important role in maintaining mitochondrial quality. Enhanced mitocytosis protects cells from mitochondrial stressor-induced loss of mitochondrial membrane potential (MMP) and mitochondrial respiration; conversely, blocking mitocytosis causes loss of MMP and mitochondrial respiration under normal conditions. Physiologically, we demonstrate that mitocytosis is required for maintaining MMP and viability in neutrophils in vivo. We propose that mitocytosis is an important mitochondrial quality-control process in migrating cells, which couples mitochondrial homeostasis with cell migration.

Metabolic regulation of innate immunity.

Immune responses are often accompanied by radical changes of cellular metabolism of immune cells. On the other hand, an ever increasing number of metabolic pathways and products have been found to possess immune regulatory functions. The field of immunometabolism that investigates the interplay between metabolism and immunity has developed rapidly during the past decade. In this chapter, we attempt to summarize the recent progresses by scientists in China on metabolic regulation of innate immunity from the following three perspectives: metabolic regulation of myeloid cell functions, metabolic adaptations of tissue resident myeloid cells, and metabolism and immunity at the mucosal surfaces.

The colonic macrophage transcription factor RBP-J orchestrates intestinal immunity against bacterial pathogens.

Macrophages play pleiotropic roles in maintaining the balance between immune tolerance and inflammatory responses in the gut. Here, we identified transcription factor RBP-J as a crucial regulator of colonic macrophage-mediated immune responses against the enteric pathogen Citrobacter rodentium. In the immune response phase, RBP-J promoted pathogen clearance by enhancing intestinal macrophage-elicited Th17 cell immune responses, which was achieved by maintenance of C/EBPß-dependent IL-6 production by overcoming miRNA-17∼92-mediated suppressive effects. RBP-J deficiency-associated phenotypes could be genetically corrected by further deleting miRNA-17∼92 in macrophages. In the late phase, noneradicated pathogens in RBP-J KO mice recruited abundant IL-1ß-expressing CD64+Ly6C+ colonic macrophages and thereby promoted persistence of ILC3-derived IL-22 to compensate for the impaired innate and adaptive immune responses, leading to ultimate clearance of pathogens. These results demonstrated that colonic macrophage-intrinsic RBP-J dynamically orchestrates intestinal immunity against pathogen infections by interfacing with key immune cells of T and innate lymphoid cell lineages.

Polarized electron acceleration in beam-driven plasma wakefield based on density down-ramp injection.

We investigate the precession of electron spins during beam-driven plasma-wakefield acceleration based on density down-ramp injection by means of full three-dimensional (3D) particle-in-cell (PIC) simulations. A relativistic electron beam generated via, e.g., laser wakefield acceleration, serves as the driving source. It traverses the prepolarized gas target and accelerates polarized electrons via the excited wakefield. We derive the criteria for the driving beam parameters and the limitation on the injected beam flux to preserve a high degree of polarization for the accelerated electrons, which are confirmed by our 3D PIC simulations and single-particle modeling. The electron-beam driver is free of the prepulse issue associated with a laser driver, thus eliminating possible depolarization of the prepolarized gas due to ionization by the prepulse. These results provide guidance for future experiments towards generating a source of polarized electrons based on wakefield acceleration.

Sweet Memories of 8 Empowered by Butyrate.

Formation of memory T cells is coupled with changes of metabolic status, yet how environmental metabolites affect the transition remains largely unknown. In this issue of Immunity, Bachem et al. (2019) report that microbiota-derived butyrate enhances the memory potential of CD8+ T cells via rewiring cellular metabolism.

Slc6a8-Mediated Creatine Uptake and Accumulation Reprogram Macrophage Polarization via Regulating Cytokine Responses.

Macrophage polarization is accompanied by drastic changes in L-arginine metabolism. Two L-arginine catalytic enzymes, iNOS and arginase 1, are well-characterized hallmark molecules of classically and alternatively activated macrophages, respectively. The third metabolic fate of L-arginine is the generation of creatine that acts as a key source of cellular energy reserve, yet little is known about the role of creatine in the immune system. Here, genetic, genomic, metabolic, and immunological analyses revealed that creatine reprogrammed macrophage polarization by suppressing M(interferon-γ [IFN-γ]) yet promoting M(interleukin-4 [IL-4]) effector functions. Mechanistically, creatine inhibited the induction of immune effector molecules, including iNOS, by suppressing IFN-γ-JAK-STAT1 transcription-factor signaling while supporting IL-4-STAT6-activated arginase 1 expression by promoting chromatin remodeling. Depletion of intracellular creatine by ablation of the creatine transporter Slc6a8 altered macrophage-mediated immune responses in vivo. These results uncover a previously uncharacterized role for creatine in macrophage polarization by modulating cellular responses to cytokines such as IFN-γ and IL-4.

Terawatt-scale optical half-cycle attosecond pulses.

Extreme-ultravoilet (XUV) attosecond pulses with durations of a few tens of attosecond have been successfully applied for exploring ultrafast electron dynamics at the atomic scale. But their weak intensities limit the further application in demonstrating nonlinear responses of inner-shell electrons. Optical attosecond pulses will provide sufficient photon flux to initiate strong-field processes. Here we proposed a novel method to generate an ultra-intense isolated optical attosecond pulse through relativistic multi-cycle laser pulse interacting with a designed gas-foil target. The underdense gas target sharpens the multi-cycle laser pulse, producing a dense layer of relativistic electrons with a thickness of a few hundred nanometers. When the dense electron layer passes through an oblique foil, it emits single ultra-intense half-cycle attosecond pulse in the visible and ultraviolet spectral range. The emitted pulse has a peak intensity exceeding 1018 W/cm2 and full-width-half-maximum duration of 200 as. The peak power of this attosecond light source reaches 2 terawatt. The proposed method relaxes the single-cycle requirement on the driving pulse for isolated attosecond pulse generation and significantly boosts the peak power, thus it may open up the route to new experiments tracking the nonlinear response of inner-shell electrons as well as nonlinear attosecond phenomena investigation.

Detergent-resistant oligomeric Leptosphaeria rhodopsin is a promising bio-nanomaterial and an alternative to bacteriorhodopsin.

Bacteriorhodopsin has attracted remarkable attention as a photoactive bio-nanomaterial in the last decades. However, its instability in the presence of detergents has restricted the extent to which bacteriorhodopsin may be applied. In this study, we investigated the oligomerization of a eukaryotic light-driven H+-pump, Leptosphaeria rhodopsin, using circular dichroism spectroscopy and other biophysical and biochemical methods. Our findings revealed that Leptosphaeria rhodopsin assembled into oligomers in the cell membrane and also in 0.05% DDM detergent micelles. Moreover, unlike bacteriorhodopsin in purple membrane, Leptosphaeria rhodopsin retained its oligomeric structure in 1% Triton X-100 and demonstrated strong resistance to other common detergents. A maximal photocurrent density of ∼85 nA/cm2 was consistently generated, which was substantially larger than that of solubilized bacteriorhodopsin (∼10 nA/cm2). Therefore, oligomeric Leptosphaeria rhodopsin may be a promising bio-nanomaterial, and an alternative to bacteriorhodopsin, especially with the use of detergents.

Coexistence of light-driven Na+ and H+ transport in a microbial rhodopsin from Nonlabens dokdonensis.

Ion pumping microbial rhodopsins are photochemically active membrane proteins, converting light energy into ion-motive-force for ATP synthesis. Nonlabens dokdonensis rhodopsin 2 (NdR2), was recently identified as a light-driven Na+ pump. However, few functional studies on NdR2 have been conducted to elucidate its mechanism of ion transport. By reconstituting NdR2 into liposomes, we proved that NdR2 functions as a light-driven Na+/H+ pump. As Na+ concentration increased, the dominant H+ pump activity switched to the Na+ pump activity at neutral pH. The inversion of pH change by the addition of CCCP at low Na+ further suggested that the transport of Na+ and H+ should coexist in NdR2. By increasing H+ concentration, the affinity for Na+ lowered, which was indicated by an increase in KM from ~31mM at pH ~7.5, to ~74mM at pH ~6.5. These results demonstrated that Na+ transport competed with H+ transport in NdR2, which was confirmed by the dominant H+ pump activity at pH ~5.7. Kinetic experiments using pyranine uncovered a transient H+ uptake, followed by an H+ release at the millisecond time scale in both Na+ and K+ solutions. Therefore, these NdR2 results may provide functional and kinetic insights into the ion transport mechanism in light-driven Na+ pumps.

Sorption of tetracycline, oxytetracycline and tylosin to eight surface sediments of Taihu Lake.

The objective of the present study is to investigate the mechanism of tetracyclines and macrolieds absorption on Taihu Lake sediments. In the study, batch technique was used to study the adsorptive behavior of three pharmaceutical antibiotics (tetracycline, oxytetracycline and tylosin) from several sediments of Taihu Lake, Zhushan Bay, Western Lakeshore, Lake Center, Southern Lakeshore, East Tai Lake, Eastern Lakeshore, Gonghu Bay and Meiliang Bay. The eight sediments showed extraordinarily high absorption affinity for all the tested antibiotics. However, especially the sediments of East Tai Lake was exceptional. The observed sorbent to solution distribution coefficient (K(d), 1 kg(-1)) was 10(2)-10(4) . The sediment of East Tai Lake showed highest organic carbon content and cation exchange capacity. A remarkably strong sorption of antibiotics to the sediment of East Tai Lake can be attributed to the cation exchange and complexation reactions between the functional groups of antibiotics and the respective charged and polar sites of the sorbents. The sorption affinity of tetracycline and oxytetracycline from the eight sediments was higher than tylosin. Tetracycline and oxytetracycline had multiple polar and ionizable functional groups. In the study within the tested pH, the zwitterion speciation is predominated; therefore, the sorption interaction (cation exchange and surface complexation) between tetracycline and sediments was expected stronger than tylosin.

Positron acceleration in a hollow plasma channel up to TeV regime.

Nowadays, human's understanding of the fundamental physics is somehow limited by the energy that our high energy accelerators can afford. Up to 4 TeV protons are realized in the Large Hadron Collider (LHC). Leptons, such as electrons and positrons, however gained energies of about 100 GeV or less. Multi-TeV lepton accelerators are still lacking due to the relatively low acceleration gradient of conventional methods, which may induce unbearable cost. On the other hand, plasmas have shown extraordinary potential in accelerating electrons and ions, providing orders of magnitude higher acceleration fields of 10-100 GV/m. In such context, we propose a plasma-based high-energy lepton accelerator, in which a weakly focusing plasma structure is formed near the beam axis. The structure preserves the emittance of the accelerated beam and produces low radiation losses. Moreover, the structure allows for a considerable decrease of the witness energy spread at the driver depletion stage.

Liquid crystalline and shear-induced properties of an aqueous solution of graphene oxide sheets.

We demonstrated here the lyotropic liquid crystalline behavior of an aqueous solution of graphene oxide (GO) sheets. Scanning electron microscope experiments revealed GO sheets self-assembled into fiber-like or sheet-like structures at different concentrations under flow conditions. As a result, the solution viscosity decreased dramatically with increasing shear stress.