A CHip off the old block.

Lineage-specific effects of upstream promoters affect ST2 expression and effector function in TH1 cells.

Promoting (and targeting) alternative expression.

Lineage-specific effects of upstream promoters affect ST2 expression and effector function in TH1cells.

Everything everywhere all at once.

In successful melanoma immunotherapy, clonal TCRs can recognize multiple tumor-specific antigens simultaneously through cross-reactivity.

Macs-imizing sepsis protection.

CD169+ macrophage-intrinsic IL-10 production mitigates mortality from sepsis.

Taking the STING out.

ABCC1 is an ATP-dependent cGAMP exporter responsible for negatively regulating STING signaling.

Distinct mechanisms of innate and adaptive immune regulation underlie poor oncologic outcomes associated with KRAS-TP53 co-alteration in pancreatic cancer.

Co-occurrent KRAS and TP53 mutations define a majority of patients with pancreatic ductal adenocarcinoma (PDAC) and define its pro-metastatic proclivity. Here, we demonstrate that KRAS-TP53 co-alteration is associated with worse survival compared with either KRAS-alone or TP53-alone altered PDAC in 245 patients with metastatic disease treated at a tertiary referral cancer center, and validate this observation in two independent molecularly annotated datasets. Compared with non-TP53 mutated KRAS-altered tumors, KRAS-TP53 co-alteration engenders disproportionately innate immune-enriched and CD8+ T-cell-excluded immune signatures. Leveraging in silico, in vitro, and in vivo models of human and murine PDAC, we discover a novel intersection between KRAS-TP53 co-altered transcriptomes, TP63-defined squamous trans-differentiation, and myeloid-cell migration into the tumor microenvironment. Comparison of single-cell transcriptomes between KRAS-TP53 co-altered and KRAS-altered/TP53WT tumors revealed cancer cell-autonomous transcriptional programs that orchestrate innate immune trafficking and function. Moreover, we uncover granulocyte-derived inflammasome activation and TNF signaling as putative paracrine mediators of innate immunoregulatory transcriptional programs in KRAS-TP53 co-altered PDAC. Immune subtyping of KRAS-TP53 co-altered PDAC reveals conflation of intratumor heterogeneity with progenitor-like stemness properties. Coalescing these distinct molecular characteristics into a KRAS-TP53 co-altered "immunoregulatory program" predicts chemoresistance in metastatic PDAC patients enrolled in the COMPASS trial, as well as worse overall survival.

Immune gains through brain drains.

In a mouse model of pneumococcal meningitis, skull channels provide extravascular signaling to the skull marrow capable of initiating local marrow hematopoiesis.

CyTOF® for the Masses.

Mass cytometry has revolutionized immunophenotyping, particularly in exploratory settings where simultaneous breadth and depth of characterization of immune populations is needed with limited samples such as in preclinical and clinical tumor immunotherapy. Mass cytometry is also a powerful tool for single-cell immunological assays, especially for complex and simultaneous characterization of diverse intratumoral immune subsets or immunotherapeutic cell populations. Through the elimination of spectral overlap seen in optical flow cytometry by replacement of fluorescent labels with metal isotopes, mass cytometry allows, on average, robust analysis of 60 individual parameters simultaneously. This is, however, associated with significantly increased complexity in the design, execution, and interpretation of mass cytometry experiments. To address the key pitfalls associated with the fragmentation, complexity, and analysis of data in mass cytometry for immunologists who are novices to these techniques, we have developed a comprehensive resource guide. Included in this review are experiment and panel design, antibody conjugations, sample staining, sample acquisition, and data pre-processing and analysis. Where feasible multiple resources for the same process are compared, allowing researchers experienced in flow cytometry but with minimal mass cytometry expertise to develop a data-driven and streamlined project workflow. It is our hope that this manuscript will prove a useful resource for both beginning and advanced users of mass cytometry.

Executive CoAching unleashes Tc22 anti-tumor capacity.

CoA-driven mitochondrial metabolism enhances the anti-tumor properties of IL-22­producing CD8+ T cells.

Turning the Tide for Academic Women in STEM: A Postpandemic Vision for Supporting Female Scientists.

The "leaky pipeline" of women in science, technology, engineering, and mathematics (STEM), which is especially acute for academic mothers, continues to be problematic as women face continuous cycles of barriers and obstacles to advancing further in their fields. The severity and prevalence of the COVID-19 pandemic both highlighted and exacerbated the unique challenges faced by female graduate students, postdocs, research staff, and principal investigators because of lockdowns, quarantines, school closures, lack of external childcare, and heightened family responsibilities, on top of professional responsibilities. This perspective provides recommendations of specific policies and practices that combat stigmas faced by women in STEM and can help them retain their careers. We discuss actions that can be taken to support women within academic institutions, journals, government/federal centers, university-level departments, and individual research groups. These recommendations are based on prior initiatives that have been successful in having a positive impact on gender equity─a central tenet of our postpandemic vision for the STEM workforce.

Ex Vivo Expansion of Th2-Polarizing Immunotherapeutic iNKT Cells from Human Peripheral Blood.

iNKT cells, classified as innate lymphocytes with invariant TCRs, have been highlighted as a putative, "off-the-shelf" cellular immunotherapeutic strategy for the treatment of malignant and nonmalignant diseases. However, their paucity in human blood limits their immunotherapeutic applications. Herein we describe a rigorously optimized 21-day ex vivo expansion method to achieve log-fold increases in immunotherapeutic human iNKT cells.

A T-rojan horse strategy for cancer immunotherapy.

CDK4/6 inhibitor-treated breast cancer cells recruit T cells via metabolic stress-induced chemokines.

B careful: Humoral responses and COVID-19 severity.

Neutralizing antibody responses to SARS-CoV-2, though often of limited longevity, have generally been assumed to be protective against COVID-19 disease.

Finding Camel-ot: A Holy Grail against pandemic SARS-CoV-2?

Engineered camelid antibody multimers can potently block SARS-CoV-2 viral entry.

Expansion and CD2/CD3/CD28 stimulation enhance Th2 cytokine secretion of human invariant NKT cells with retained anti-tumor cytotoxicity.

BACKGROUND AIMS: Key obstacles in human iNKT cell translational research and immunotherapy include the lack of robust protocols for dependable expansion of human iNKT cells and the paucity of data on phenotypes in post-expanded cells. METHODS: We delineate expansion methods using interleukin (IL)-2, IL-7 and allogeneic feeder cells and anti-CD2/CD3/CD28 stimulation by which to dependably augment Th2 polarization and direct cytotoxicity of human peripheral blood CD3+Vα24+Vß11+ iNKT cells. RESULTS: Gene and protein expression profiling demonstrated augmented Th2 cytokine secretion (IL-4, IL-5, IL-13) in expanded iNKT cells stimulated with anti-CD2/CD3/CD28 antibodies. Cytotoxic effector molecules including granzyme B were increased in expanded iNKT cells after CD2/CD3/CD28 stimulation. Direct cytotoxicity assays using unstimulated expanded iNKT cell effectors revealed α-galactosyl ceramide (α-GalCer)-dependent killing of the T-ALL cell line Jurkat. Moreover, CD2/CD3/CD28 stimulation of expanded iNKT cells augmented their (α-GalCer-independent) killing of Jurkat cells. Co-culture of expanded iNKT cells with stimulated responder cells confirmed contact-dependent inhibition of activated CD4+ and CD8+ responder T cells. DISCUSSION: These data establish a robust protocol to expand and novel pathways to enhance Th2 cytokine secretion and direct cytotoxicity in human iNKT cells, findings with direct implications for autoimmunity, vaccine augmentation and anti-infective immunity, cancer immunotherapy and transplantation.

Isolation of Lamina Propria Mononuclear Cells from Murine Colon Using Collagenase E.

The intestine is the home to the largest number of immune cells in the body. The small and large intestinal immune systems police exposure to exogenous antigens and modulate responses to potent microbially derived immune stimuli. For this reason, the intestine is a major target site of immune dysregulation and inflammation in many diseases including but, not limited to inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, graft-versus-host disease (GVHD) after bone marrow transplantation (BMT), and many allergic and infectious conditions. Murine models of gastrointestinal inflammation and colitis are heavily used to study GI complications and to pre-clinically optimize strategies for prevention and treatment. Data gleaned from these models via isolation and phenotypic analysis of immune cells from the intestine is critical to further immune understanding that can be applied to ameliorate gastrointestinal and systemic inflammatory disorders. This report describes a highly effective protocol for the isolation of mononuclear cells (MNC) from the colon using a mixed silica-based density gradient interface. This method reproducibly isolates a significant number of viable leukocytes while minimizing contaminating debris, allowing subsequent immune phenotyping by flow cytometry or other methods.