Cancer Biology


    Protocols in Current Issue
    CRISPR/Cas9-mediated Gene Knockout Followed by Negative Selection Leads to a Complete TCR Depletion in orthoCAR19 T Cells

    Genome-editing technologies, especially CRISPR (clustered regularly interspaced short palindrome repeats)/Cas9 (CRISPR-associated protein 9), endows researchers the ability to make efficient, simple, and precise genomic DNA changes in many eukaryotic cell types. CRISPR/Cas9-mediated efficient gene knockout holds huge potential to improve the

    Measurement of Cell Intrinsic TGF-β Activation Mediated by the Integrin αvβ8
    Authors:  Robert Ian Seed and Stephen Lloyd Nishimura, date: 04/20/2022, view: 1571, Q&A: 0

    Transforming growth factor beta (TGF-β) is a multi-functional cytokine that plays a significant role in multiple diseases, including fibrosis and tumor progression. Whilst the biologic effects of TGF-β are well characterized, it is unclear how TGF-β signaling is regulated to impart specific responses within certain cell types. One

    An Alternative Technique for Monitoring the Live Interaction of Monocytes and Tumor Cells with Nanoparticles in the Mouse Lung
    Authors:  Fernanda Ramos-Gomes, Nathalia Ferreira, Frauke Alves and M. Andrea Markus, date: 01/20/2022, view: 1480, Q&A: 0

    Nanomaterials are increasingly used for the diagnosis and treatment of cancer, including lung cancer. For the clinical translation of nano-based theranostics, it is vital to detect and monitor their accumulation in the tumor, as well as their interaction with tumor, immune cells, and the tumor microenvironment (TME). While high resolution

    Multi-color Flow Cytometry for Comprehensive Analysis of the Tumor Immune Infiltrate in a Murine Model of Breast Cancer
    Authors:  Ana S. Almeida, Miriam R. Fein and Mikala Egeblad, date: 06/05/2021, view: 3532, Q&A: 0

    Flow cytometry is a popular laser-based technology that allows the phenotypic and functional characterization of individual cells in a high-throughput manner. Here, we describe a detailed procedure for preparing a single-cell suspension from mammary tumors of the mouse mammary tumor virus-polyoma middle T (MMTV-PyMT) and analyzing these cells by

    In vitro STING Activation with the cGAMP-STINGΔTM Signaling Complex
    Authors:  Yanpu He, Celestine Hong, Darrell J. Irvine, Jiahe Li and Paula T. Hammond, date: 02/05/2021, view: 3718, Q&A: 0
    [Abstract] Activating the STING (stimulator of interferon genes) signaling pathway via administration of STING agonist cyclic GMP-AMP (cGAMP) has shown great promise in cancer immunotherapy. While state-of-the-art approaches have predominantly focused on the encapsulation of cGAMP into liposomes or polymersomes for cellular delivery, we discovered that the ...
    A 3D Skin Melanoma Spheroid-Based Model to Assess Tumor-Immune Cell Interactions
    Authors:  Marek Wagner and Shigeo Koyasu, date: 12/05/2020, view: 2488, Q&A: 0
    [Abstract] Three-dimensional (3D) tumor spheroids have the potential to bridge the gap between two-dimensional (2D) monolayer tumor cell cultures and solid tumors with which they share a significant degree of similarity. However, the progression of solid tumors is often influenced by the dynamic and reciprocal interactions between tumor and immune cells. ...
    Generation and Testing of Fluorescent Adaptable Simple Theranostic (FAST) Proteins
    [Abstract] This protocol provides a step-by-step method to create recombinant fluorescent fusion proteins that can be secreted from mammalian cell lines. This builds on many other recombinant protein and fluorescent protein techniques, but is among the first to harness fluorescent fusion proteins secreted directly into cell culture supernatant. This opens ...
    Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)
    Authors:  Magnez Romain, Bryan Thiroux, Morgane Tardy, Bruno Quesnel and Xavier Thuru, date: 04/05/2020, view: 4928, Q&A: 0
    [Abstract] The binding interactions of PD-1 and PD-L1 have been studied by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but these investigations resulted in controversy regarding the values of the dissociation constant (Kd) (Freeman et al., 2000). MST is a powerful new method for the ...
    Evaluation of Mucosal and Systemic Vaccine Responses by Cyclic di-GMP (CDG)-adjuvanted Protein Subunit Vaccines
    Authors:  Samira Mansouri and Lei Jin, date: 04/20/2019, view: 3928, Q&A: 0
    [Abstract] Intranasal administration of vaccine adjuvants directly deliver therapeutic agents to the lungs to induce potent lung mucosal immune responses. Cyclic di-GMP (CDG) is a promising mucosal vaccine adjuvant candidate capable of inducing protective immunity. This protocol describes an in vivo approach to induce and detect mucosal (lung) and ...
    In situ, Cell-free Protein Expression on Microarrays and Their Use for the Detection of Immune Responses
    [Abstract] Until recently, whole-proteome microarrays for comprehensive studies of protein interactions were mostly produced by individual cloning and cellular expression of very many open reading frames, followed by protein isolation and purification as well as array production. To overcome this cumbersome process, we have developed a method to generate ...

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