How to make a better T cell: in vivo CRISPR screens have some answers

Understanding what regulates CD8+ T cell responses is key to effectively harnessing these cells in human disease. In this issue of Cell, Huang et al. and Chen et al. use in vivo CRISPR screens to discover novel regulators of CD8+ T cell immunity to engineer a more efficacious response against cancer and infections

Contrast-enhanced ultrasound identifies early extrahepatic collateral contributing to residual hepatocellular tumor viability after transarterial chemoembolization.

The mainstay of treatment for unresectable hepatocellular carcinoma is locoregional therapy including percutaneous ablation and transarterial chemo- and radioembolization. While monitoring for tumor response after transarterial chemoembolization is crucial, current imaging strategies are suboptimal. The standard of care is contrast-enhanced magnetic resonance imaging or computed tomography imaging performed at least 4 to 6 weeks after therapy. We present a case in which contrast-enhanced ultrasound identified a specific extra-hepatic collateral from the gastroduodenal artery supplying residual viable tumor and assisting with directed transarterial management

Evaluation of Hepatocellular Carcinoma Transarterial Chemoembolization using Quantitative Analysis of 2D and 3D Real-time Contrast Enhanced Ultrasound.

Quantitative 2D and 3D contrast-enhanced ultrasound (CEUS) was assessed to evaluate early transarterial chemoembolization (TACE) treatment response. Seventeen patients scheduled for TACE for the treatment of hepatocellular carcinoma participated in the study. 2D and 3D CEUS were performed for each patient at three time points: Prior to TACE, 1-2 weeks post TACE, and 1 month post TACE. Peak-intensities of the tumor and surrounding liver tissue were calculated from 2D and 3D data before and after TACE and used to evaluate tumor treatment response. Residual tumor percentages were calculated from 2D and 3D CEUS acquired 1-2 weeks and 1 month post TACE and compared with results from MRI 1 month post TACE. Nine subjects had complete response while 8 had incomplete response. Peak-intensities of the tumor from 3D CEUS prior to TACE were similar between the complete and incomplete treatment groups (p = 0.70), while 1-2 weeks (p \u3c 0.01) and 1 month post treatment (p \u3c 0.01) were significantly lower in the complete treatment group than in the incomplete treatment group. For 2D CEUS, only the peak-intensity values of the tumor from 1 month post TACE were significantly different (p \u3c 0.01). The correlation coefficients between 2D and 3D residual tumor estimates 1-2 weeks post TACE and the estimates from MRI were 0.73 and 0.94, respectively, while those from 2D and 3D CEUS 1 month post TACE were 0.66 and 0.91, respectively. Quantitative analysis on 2D and 3D CEUS shows potential to differentiate patients with complete versus incomplete response to TACE as early as 1-2 weeks post treatment

12PLECTIN GALACTOSIDE-BINDING SOLUBLE 3 BINDING PROTEIN (LGALS3BP) IS A CANCER-ASSOCIATED LIGAND FOR INHIBITORY SIGLECS

Tumor cells subvert the control of the immune system by downregulation of their antigenicity and production of an immunosuppressive microenvironment including the upregulation and engagement of inhibitory receptors on immune cells. Therapeutic strategies have demonstrated that the immune system can be reactivated and control established cancers by blocking inhibitory receptors on immune cells such CTLA-4 and PD1. While such activation of the immune system is successful in some patients, many patients still show cancer progression after some time. Thus, the definition of new targetable immunomodulatory pathways is needed to improve the outcome in those patients. Recent evidence suggests that sialic acid dependent ligands on tumor cells can engage inhibitory sialic acid binding immunoglobulin-like lectins (Siglecs) on NK cells and cells of the myelomonocytic lineage and thereby facilitate evasion of immune-mediated killing. Moreover, the presence of a natural variant of Siglec-9 with reduced binding capacity to sialic acid dependent ligands in patients with non-small cell lung cancer improved the two year survival in a retrospective multivariate analysis. Here we identify a novel cancer-associated ligand for immuno-inhibitory Siglecs by affinity chromatography and subsequent proteomic analysis. LectinGalactoside-Binding Soluble 3 Binding Protein (LGALS3BP) bound to various inhibitory Siglecs including Siglec-5, Siglec-9 and Siglec-10 with high affinity. LGALS3BP was previously found to be upregulated in various carcinomas such as breast, colorectal, prostate and lung cancer and linked to advanced stage and poor prognosis. The exact function during cancer progression, however, was not yet defined. Our findings provide a novel insight into how LGALS3BP could promote immune evasion by inhibiting immune cell activation through engagement of Siglecs and defines LGALS3BP-Siglec interactions as potential novel target to interfere with cancer progression and reactivate the immune system against carcinomas. Disclosure: All authors have declared no conflicts of interes

Parallel APSM for Fast and Adaptive Digital SIC in Full-Duplex Transceivers with Nonlinearity

This paper presents a kernel-based adaptive filter that is applied for the digital domain self-interference cancellation (SIC) in a transceiver operating in full-duplex (FD) mode. In FD, the benefit of simultaneous transmission and receiving of signals comes at the price of strong self-interference (SI). In this work, we are primarily interested in suppressing the SI using an adaptive filter namely adaptive projected subgradient method (APSM) in a reproducing kernel Hilbert space (RKHS) of functions. Using the projection concept as a powerful tool, APSM is used to model and consequently remove the SI. A low-complexity and fast-tracking algorithm is provided taking advantage of parallel projections as well as the kernel trick in RKHS. The performance of the proposed method is evaluated on real measurement data. The method illustrates the good performance of the proposed adaptive filter, compared to the known popular benchmarks. They demonstrate that the kernel-based algorithm achieves a favorable level of digital SIC while enabling parallel computation-based implementation within a rich and nonlinear function space, thanks to the employed adaptive filtering method

Genomic insights into the rapid emergence and evolution of MDR in Staphylococcus pseudintermedius.

OBJECTIVES: MDR methicillin-resistant Staphylococcus pseudintermedius (MRSP) strains have emerged rapidly as major canine pathogens and present serious treatment issues and concerns to public health due to their, albeit low, zoonotic potential. A further understanding of the genetics of resistance arising from a broadly susceptible background of S. pseudintermedius is needed. METHODS: We sequenced the genomes of 12 S. pseudintermedius isolates of varied STs and resistance phenotypes. RESULTS: Nine distinct clonal lineages had acquired either staphylococcal cassette chromosome (SCC) mec elements and/or Tn5405-like elements carrying up to five resistance genes [aphA3, sat, aadE, erm(B), dfrG] to generate MRSP, MDR methicillin-susceptible S. pseudintermedius and MDR MRSP populations. The most successful and clinically problematic MDR MRSP clones, ST68 SCCmecV(T) and ST71 SCCmecII-III, have further accumulated mutations in gyrA and grlA conferring resistance to fluoroquinolones. The carriage of additional mobile genetic elements (MGEs) was highly variable, suggesting that horizontal gene transfer is frequent in S. pseudintermedius populations. CONCLUSIONS: Importantly, the data suggest that MDR MRSP evolved rapidly by the acquisition of a very limited number of MGEs and mutations, and that the use of many classes of antimicrobials may co-select for the spread and emergence of MDR and XDR strains. Antimicrobial stewardship will need to be comprehensive, encompassing human medicine and veterinary disciplines to successfully preserve antimicrobial efficacy

IL-33 expression in response to SARS-CoV-2 correlates with seropositivity in COVID-19 convalescent individuals

Our understanding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still developing. We perform an observational study to investigate seroprevalence and immune responses in subjects professionally exposed to SARS-CoV-2 and their family members (155 individuals; ages 5-79 years). Seropositivity for SARS-CoV-2 Spike glycoprotein aligns with PCR results that confirm the previous infection. Anti-Spike IgG/IgM titers remain high 60 days post-infection and do not strongly associate with symptoms, except for fever. We analyze PBMCs from a subset of seropositive and seronegative adults. TLR7 agonist-activation reveals an increased population of IL-6+TNF-IL-1β+ monocytes, while SARS-CoV-2 peptide stimulation elicits IL-33, IL-6, IFNa2, and IL-23 expression in seropositive individuals. IL-33 correlates with CD4+ T cell activation in PBMCs from convalescent subjects and is likely due to T cell-mediated effects on IL-33-producing cells. IL-33 is associated with pulmonary infection and chronic diseases like asthma and COPD, but its role in COVID-19 is unknown. Analysis of published scRNAseq data of bronchoalveolar lavage fluid (BALF) from patients with mild to severe COVID-19 reveals a population of IL-33-producing cells that increases with the disease. Together these findings show that IL-33 production is linked to SARS-CoV-2 infection and warrant further investigation of IL-33 in COVID-19 pathogenesis and immunity

Sulfur sequestration promotes multicellularity during nutrient limitation

The behaviour of Dictyostelium discoideum depends on nutrients. When sufficient food is present these amoebae exist in a unicellular state, but upon starvation they aggregate into a multicellular organism. This biology makes D. discoideum an ideal model for investigating how fundamental metabolism commands cell differentiation and function. Here we show that reactive oxygen species-generated as a consequence of nutrient limitation-lead to the sequestration of cysteine in the antioxidant glutathione. This sequestration limits the use of the sulfur atom of cysteine in processes that contribute to mitochondrial metabolism and cellular proliferation, such as protein translation and the activity of enzymes that contain an iron-sulfur cluster. The regulated sequestration of sulfur maintains D. discoideum in a nonproliferating state that paves the way for multicellular development. This mechanism of signalling through reactive oxygen species highlights oxygen and sulfur as simple signalling molecules that dictate cell fate in an early eukaryote, with implications for responses to nutrient fluctuations in multicellular eukaryotes

Dynamic Cardiolipin Synthesis Is Required for CD8⁺ T Cell Immunity

Mitochondria constantly adapt to the metabolic needs of a cell. This mitochondrial plasticity is critical to T cells, which modulate metabolism depending on antigen-driven signals and environment. We show here that de novo synthesis of the mitochondrial membrane-specific lipid cardiolipin maintains CD8+ T cell function. T cells deficient for the cardiolipin-synthesizing enzyme PTPMT1 had reduced cardiolipin and responded poorly to antigen because basal cardiolipin levels were required for activation. However, neither de novo cardiolipin synthesis, nor its Tafazzin-dependent remodeling, was needed for T cell activation. In contrast, PTPMT1-dependent cardiolipin synthesis was vital when mitochondrial fitness was required, most notably during memory T cell differentiation or nutrient stress. We also found CD8+ T cell defects in a small cohort of patients with Barth syndrome, where TAFAZZIN is mutated, and in a Tafazzin-deficient mouse model. Thus, the dynamic regulation of a single mitochondrial lipid is crucial for CD8+ T cell immunity