Image_3_Loss-of-Function in SMAD4 Might Not Be Critical for Human Natural Killer Cell Responsiveness to TGF-β.TIF

We characterized the NK cell phenotype and function in three family members with Hereditary Hemorrhagic Telangiectasia (HHT) due to heterozygous SMAD4 mutations. Loss-of-function mutation in this gene did not induce developmental effects to alter CD56bright or CD56dim NK cell subset proportions in peripheral blood; and did not result in major differences in either their IL-15-induced proliferation, or their cytokine secretion response to TGF-β1. These data suggest that SMAD4 plays a redundant role in downstream TGF-β signaling in NK cells.</p

Image_1_Loss-of-Function in SMAD4 Might Not Be Critical for Human Natural Killer Cell Responsiveness to TGF-β.TIFF

We characterized the NK cell phenotype and function in three family members with Hereditary Hemorrhagic Telangiectasia (HHT) due to heterozygous SMAD4 mutations. Loss-of-function mutation in this gene did not induce developmental effects to alter CD56bright or CD56dim NK cell subset proportions in peripheral blood; and did not result in major differences in either their IL-15-induced proliferation, or their cytokine secretion response to TGF-β1. These data suggest that SMAD4 plays a redundant role in downstream TGF-β signaling in NK cells.</p

Figure S1 from Neoadjuvant Interferons: Critical for Effective PD-1–Based Immunotherapy in TNBC

Varying doses of IFN treatment and PD-L1/IFNAR expression</p

Figure S2 from Neoadjuvant Interferons: Critical for Effective PD-1–Based Immunotherapy in TNBC

The effect of poly(I:C) on immune cell activation</p

Figure S3 from Neoadjuvant Interferons: Critical for Effective PD-1–Based Immunotherapy in TNBC

The effect of combination treatment on T cell activation</p

Data_Sheet_1_NK Cell Priming From Endogenous Homeostatic Signals Is Modulated by CIS.PDF

Natural killer (NK) cell activation is controlled by a balance of activating and inhibitory signals and cytokines such as IL-15. We previously identified cytokine-inducible SH2-containing protein (CIS) as a negative regulator of IL-15 signaling in NK cells under inflammatory conditions. While the functional effect of Cish-deficiency in NK cells was obvious by their increased anti-tumor immunity and hyper-proliferative response to IL-15, it remained unclear how CIS regulates NK cell biology in steady-state. Here, we investigated the role of CIS in the homeostatic maintenance of NK cells and found CIS-ablation promoted terminal differentiation of NK cells and increased turnover, suggesting that under steady-state conditions, CIS plays a role in maintaining IL-15 driven regulation of NK cells in vivo. However, hyper-responsiveness to IL-15 did not manifest in NK cell accumulation, even when the essential NK cell apoptosis mediator, Bcl2l11 (BIM) was deleted in addition to Cish. Instead, loss of CIS conferred a lower activation threshold, evidenced by augmented functionality on a per cell basis both in vitro and in vivo without prior priming. We conclude that Cish regulates IL-15 signaling in NK cells in vivo, and through the rewiring of several activation pathways leads to a reduction in activation threshold, decreasing the requirement for priming and improving NK cell anti-tumor function. Furthermore, this study highlights the tight regulation of NK cell homeostasis by several pathways which prevent NK cell accumulation when IL-15 signaling and intrinsic apoptosis are dysregulated.</p

Image_1_GM-CSF Quantity Has a Selective Effect on Granulocytic vs. Monocytic Myeloid Development and Function.PDF

GM-CSF promotes myeloid differentiation of cultured bone marrow cells into cells of the granulocytic and monocytic lineage; the latter can further differentiate into monocytes/macrophages and dendritic cells. How GM-CSF selects for these different myeloid fates is unresolved. GM-CSF levels can change either iatrogenically (e.g., augmenting leukopoiesis after radiotherapy) or naturally (e.g., during infection or inflammation) resulting in different immunological outcomes. Therefore, we asked whether the dose of GM-CSF may regulate the development of three types of myeloid cells. Here, we showed that GM-CSF acted as a molecular rheostat where the quantity determined which cell type was favored; moreover, the cellular process by which this was achieved was different for each cell type. Thus, low quantities of GM-CSF promoted the granulocytic lineage, mainly through survival. High quantities promoted the monocytic lineage, mainly through proliferation, whereas moderate quantities promoted moDCs, mainly through differentiation. Finally, we demonstrated that monocytes/macrophages generated with different doses of GM-CSF differed in function. We contend that this selective effect of GM-CSF dose on myeloid differentiation and function should be taken into consideration during pathophysiological states that may alter GM-CSF levels and during GM-CSF agonistic or antagonistic therapy.</p

Supplementary figure legends from Neoadjuvant Interferons: Critical for Effective PD-1–Based Immunotherapy in TNBC

Figure legends for supplementary figures 1, 2 & 3</p

Supplementary Figures S1-S6 from Targeting Adenosine in BRAF-Mutant Melanoma Reduces Tumor Growth and Metastasis

This file contains Supplementary Figure S1A-B, Figure S2A, Figure S3-F, Figure S4A-B, Figure S5A-E and Figure S6A. Figure S1. CD73 expression in melanoma does not associate with survival. Figure S2. Hematopoietic A2AR-deficiency reduces inducible BRAF-mutated tumor growth. Figure S3. A2AR inhibition enhances CD8+ T and NK cell infiltration into the tumor microenvironment. Figure S4. SOX10 expression is present in skin and BRAF-mutated, PTENdeficient tumors. Figure S5. A2AR and BRAF inhibition in combination provides improved tumor control. Figure S6. BRAF and MEK downregulate tumor-derived CD73 expression.</p

Supplementary Figure 2 from Loss of Host Type-I IFN Signaling Accelerates Metastasis and Impairs NK-cell Antitumor Function in Multiple Models of Breast Cancer

IL-2 stimulated NK cells derived from Ifnar1-/- or WT mice have comparable cytotoxicity against syngeneic breast tumor lines.</p