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Fig.6.B showing Flow cytometry/Cell sorting from the publication: CLDN18.2 and 4-1BB bispecific antibody givastomig exerts antitumor activity through CLDN18.2-expressing tumor-directed T-cell activation.

Fig.3.H showing Western Blotting from the publication: A20 and ABIN-1 cooperate in balancing CBM complex-triggered NF-κB signaling in activated T cells.

Fig.3.C showing Western Blotting from the publication: A20 and ABIN-1 cooperate in balancing CBM complex-triggered NF-κB signaling in activated T cells.

Fig.4.K showing Western Blotting from the publication: A20 and ABIN-1 cooperate in balancing CBM complex-triggered NF-κB signaling in activated T cells.

Fig.6.E showing Western Blotting from the publication: A20 and ABIN-1 cooperate in balancing CBM complex-triggered NF-κB signaling in activated T cells.

Fig.5.A showing Flow cytometry/Cell sorting in a Mus musculus (House mouse) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.5.B showing Flow cytometry/Cell sorting in a Mus musculus (House mouse) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.5.E showing Western Blotting in a Homo sapiens (Human) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.5.F showing Western Blotting in a Homo sapiens (Human) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.8.B showing Flow cytometry/Cell sorting in a Mus musculus (House mouse) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.8.H showing Flow cytometry/Cell sorting in a Mus musculus (House mouse) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.8.G showing Flow cytometry/Cell sorting in a Mus musculus (House mouse) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.8.A showing Flow cytometry/Cell sorting in a Mus musculus (House mouse) sample from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.7.A showing Flow cytometry/Cell sorting from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.7.C showing Flow cytometry/Cell sorting from the publication: SNX27 links DGKζ to the control of transcriptional and metabolic programs in T lymphocytes.

Fig.1.C showing Western Blotting from the publication: CKIP-1 is an intrinsic negative regulator of T-cell activation through an interaction with CARMA1.

16 images found

RIG-I Stimulation Enhances Effector Function and Proliferation of Primary Human CD8 T Cells

Preprint on BioRxiv : the Preprint Server for Biology on 29 January 2025 by Mohamed, A. A., Wallerath, C., et al.

Cytotoxic CD8 T lymphocytes (CTL) are crucial in antiviral immune responses. However, their recruitment to infection sites renders them at risk of viral infection that could modulate their effector activity. CTL express RIG-I that detects cytosolic viral RNA and subsequently induce antiviral gene expression. Here, we investigated how influenza A virus (IAV) infection influence TCR-dependent effector responses. IAV infection of CTL stimulated RIG-I, and activated downstream pathways including TBK1 and NF-ϰB, resulting in type-I interferon secretion. Transfection of CTL with a pure RIG-I ligand, tri-phosphorylated double stranded RNA(3p-dsRNA), not only stimulated these pathways but also enhanced CTL proliferation in vitro and protected them from IAV infection. Analogous with a positive effect on CD8 effector function, both IAV infection and RIG-I ligand transfection enhanced CTL degranulation and cytokine secretion. Conversely, activation of CTL via CD3/CD28-crosslinking increased their susceptibility to IAV infection. Altogether, RIG-I stimulation either by IAV infection or 3p-dsRNA transfection promoted cell intrinsic antiviral pathways and enhanced CD8 effector functions. These findings suggest that RIG-I agonists could enhance and prolong CTL effector function in immunotherapy.

SLC7A5 regulates tryptophan uptake and PD‑L1 expression levels via the kynurenine pathway in ovarian cancer.

In Oncology Letters on 1 January 2025 by Jiang, R., Jin, B., et al.

Ovarian cancer is the third most common gynecological malignancy worldwide and the fifth leading cause of cancer-related death among women. This may be attributed to difficulties in diagnosing early-stage ovarian cancer, as it is typically asymptomatic until metastases, and due to the ineffective management of patients with late-stage ovarian cancer. The aim of the present study was to investigate potential therapeutic targets for the treatment of ovarian cancer. Bioinformatics techniques were used to analyze the expression levels of tryptophan (Trp) metabolism-related genes in tissue samples from patients with ovarian cancer. Additionally, western blots, clonogenic assays, immunohistochemical staining, chromatin immunoprecipitation-quantitative PCR, cell co-culture assays, a xenograft model and high-performance liquid chromatography-tandem mass spectrometry were performed to evaluate the antitumor effects of genes identified from the bioinformatics analysis. Increased expression levels of the amino acid transporter, solute carrier family 7 member 5 (SLC7A5), in tissue samples from patients with ovarian cancer was demonstrated. Inhibition of SLC7A5 reduced ovarian cancer cell proliferation through G2/M cell cycle arrest and blocked intracellular aryl hydrocarbon receptor nucleus entry, which downregulated PD-L1 expression levels. Dysregulation of Trp metabolism in ovarian cancer tissue samples, as well as the upregulation of kynurenine expression levels in the plasma of patients with ovarian cancer, were demonstrated to be unfavorable prognostic factors for the progression-free survival of patients with ovarian cancer. The present study demonstrated that the dysregulation of Trp metabolism could potentially be used as a diagnostic biomarker for ovarian cancer, as well as the potential of targeting SLC7A5 for immunotherapeutic management of patients with ovarian cancer in the future.
Copyright: © 2024 Jiang et al.

CD97 maintains tumorigenicity of glioblastoma stem cells via mTORC2 signaling and is targeted by CAR Th9 cells.

In Cell Reports Medicine on 17 December 2024 by Zhou, S., Lin, W., et al.

Glioblastoma (GBM) stem cells (GSCs) contribute to poor prognosis in patients with GBM. Identifying molecular markers is crucial for developing targeted therapies. Here, we identify cluster of differentiation 97 (CD97) as an optimal GSC surface antigen for potential targeting by chimeric antigen receptor (CAR) T cell therapy through in vitro antibody screening. CD97 is consistently expressed in all validated patient-derived GSCs and positively correlated with known intracellular GSC markers. Silencing CD97 reduces GSC tumorigenicity-related activities, including self-renewal, proliferation, and tumor progression. Transcriptome analysis reveals that CD97 activates mTORC2, leading to AKT S473 phosphorylation and enhanced expression of the downstream genes ARHGAP1, BZW1, and BZW2. Inhibiting mTORC2 with JR-AB2-011 suppresses GSC tumorigenicity and downstream gene expression. We develop CD97-CAR T helper (Th) 9 cells, which exhibit potent cytotoxic effects in vitro and extend survival in mice. These findings suggest that CD97 is a promising GSC-enriched antigen and that targeting it with CAR Th9 cells offers a potential therapeutic strategy for GBM.
Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.

AMPK activation induces RALDH+ tolerogenic dendritic cells by rewiring glucose and lipid metabolism.

In The Journal of Cell Biology on 7 October 2024 by Brombacher, E. C., Patente, T. A., et al.

Dendritic cell (DC) activation and function are underpinned by profound changes in cellular metabolism. Several studies indicate that the ability of DCs to promote tolerance is dependent on catabolic metabolism. Yet the contribution of AMP-activated kinase (AMPK), a central energy sensor promoting catabolism, to DC tolerogenicity remains unknown. Here, we show that AMPK activation renders human monocyte-derived DCs tolerogenic as evidenced by an enhanced ability to drive differentiation of regulatory T cells, a process dependent on increased RALDH activity. This is accompanied by several metabolic changes, including increased breakdown of glycerophospholipids, enhanced mitochondrial fission-dependent fatty acid oxidation, and upregulated glucose catabolism. This metabolic rewiring is functionally important as we found interference with these metabolic processes to reduce to various degrees AMPK-induced RALDH activity as well as the tolerogenic capacity of moDCs. Altogether, our findings reveal a key role for AMPK signaling in shaping DC tolerogenicity and suggest AMPK as a target to direct DC-driven tolerogenic responses in therapeutic settings.
© 2024 Brombacher et al.

Alternative 3'UTR expression induced by T cell activation is regulated in a temporal and signal dependent manner.

In Scientific Reports on 14 May 2024 by Blake, D., Gazzara, M. R., et al.

The length of 3' untranslated regions (3'UTR) is highly regulated during many transitions in cell state, including T cell activation, through the process of alternative polyadenylation (APA). However, the regulatory mechanisms and functional consequences of APA remain largely unexplored. Here we present a detailed analysis of the temporal and condition-specific regulation of APA following activation of primary human CD4+ T cells. We find that global APA changes are regulated temporally and CD28 costimulatory signals enhance a subset of these changes. Most APA changes upon T cell activation involve 3'UTR shortening, although a set of genes enriched for function in the mTOR pathway exhibit 3'UTR lengthening. While upregulation of the core polyadenylation machinery likely induces 3'UTR shortening following prolonged T cell stimulation; a significant program of APA changes occur prior to cellular proliferation or upregulation of the APA machinery. Motif analysis suggests that at least a subset of these early changes in APA are driven by upregulation of RBM3, an RNA-binding protein which competes with the APA machinery for binding. Together this work expands our understanding of the impact and mechanisms of APA in response to T cell activation and suggests new mechanisms by which APA may be regulated.
© 2024. The Author(s).

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A20 and ABIN-1 cooperate in balancing CBM complex-triggered NF-κB signaling in activated T cells.

In Cell Mol Life Sci on 31 January 2022 by Yin, H., Karayel, Ö., et al.

Fig.4.K

Interdependency of ABIN-1 and A20 in counteracting CBM signaling in Jurkat T cells. A, B BCL10 IP and detection of BCL10-bound CARD11, ABIN-1 and A20 in parental, ABIN-1 KO and A20 KO Jurkat T cells after P/I stimulation analyzed by Western blot. C, D ABIN-1 and A20 expression after viral transduction in parental and KO Jurkat T cells analyzed by Western blot. E, F NF-κB-EGFP reporter activation in transduced ABIN-1 KO or A20 KO Jurkat T cells following CD3/CD28 stimulation (1–6 h) analyzed by flow cytometry. Quantification was done by calculating median fluorescence intensity (MFI). Data represent means ± SEM (n = 4) (E) or (n = 3) (F) and quantification was done by two-way ANOVA with Dunnett’s multiple comparisons test, compared with Mock group. *p < 0.05, **p < 0.01, ****p < 0.0001. G, H Active MALT1 was detected in transduced ABIN-1 KO or A20 KO Jurkat T cells after CD3/CD28 stimulation by MALT1-ABP PD assay and Western blot. I, K Cleavage of CYLD and HOIL-1 following 2 h stimulation with CD3/CD28 in ABIN-1 KO (I) or A20 KO (K) Jurkat T cells after viral rescue/expression cells was determined by Western blot. J, L The ratio of cleaved to full-length CYLD and HOIL-1 was measured by densitometric analyses to quantify the effect of ABIN-1 or A20 reconstitution/expression on MALT1 substrate cleavage shown in I and K. Data represent mean ± SEM (n = 4) (J) or (n = 5) (L) and quantification was done by two-way ANOVA with Dunnett’s multiple comparisons test, compared with Mock group. ns non-significant, *p < 0.05, ***p < 0.001, ****p < 0.0001 less

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CKIP-1 is an intrinsic negative regulator of T-cell activation through an interaction with CARMA1.

In PLoS One on 28 January 2014 by Sakamoto, T., Kobayashi, M., et al.

Fig.1.C

Identification of CKIP-1 as a negative regulator in NF-κB activation.(A) 400 pmol of human CKIP-1-specific siRNA or non-targeting siRNA together with 5 µg of κB-Luc, 0.1 µg of Renilla-Luc were electroporated into Jurkat T cells. Luciferase activity was assayed after 48 hr. The reduction of endogenous CKIP-1 protein levels was analyzed by Western blotting. (B) Jurkat T cells were electroporated with human CKIP-1-specific siRNA or non-targeting siRNA using AMAXA Nucleofector System (Lonza). Thirty hours later, nuclear protein extracts were harvested and NF-κB activity was measured by TransAM NF-κB p65 chemi kit (Active Motif). The reduction of endogenous CKIP-1 protein levels was analyzed by Western blotting. (C) Jurkat T cells were transfected with 5 µg of CKIP-1 or empty vector (mock) together with 5 µg of κB-Luc and 0.1 µg of Renilla-Luc. Nineteen hours later, cells were stimulated for 5 hr upon CD3 (2 µg/ml), CD3/CD28 (2 µg/ml each), TNFα (20 ng/ml), PMA (10 ng/ml) or PMA (10 ng/ml) + CD28 (2 µg/ml). The expressed protein levels were analyzed by Western blotting. (D) Jurkat T cells were transfected with 5 µg of CKIP-1 or empty vector (mock). Twenty-four hours later, cells were stimulated for 30 min upon PMA (10 ng/ml). Then cells were harvested and NF-κB activity was measured by TransAM NF-κB p65 chemi kit. The expressed protein levels were analyzed by Western blotting. Values represent the average of three independent experiments and error bars represent the SD from the average. less

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