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T Cell Activation and Clonal Selection01:22

T Cell Activation and Clonal Selection

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T cells are integral to our adaptive immune system, recognizing and effectively responding to foreign antigens. T cell activation and clonal selection are pivotal in orchestrating this immune response. This article elucidates these mechanisms, detailing the roles of cluster of differentiation (CD) markers, major histocompatibility complex (MHC) molecules, costimulatory signals, and the process of clonal selection.
Naive T cells that have not yet encountered an antigen express two primary CD...
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Protein Complex Assembly02:41

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Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...
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Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

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Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
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B Cell Activation and Differentiation01:24

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The adaptive immune response, a sophisticated defense mechanism, relies on the activation and differentiation of B lymphocytes, or B cells. These processes enable our bodies to mount a tailored response against specific pathogens such as bacteria, free virus particles, toxins, and parasites.
When naive B cells encounter a specific antigen that can bind to the B cell receptor (BCR) on their surface, they undergo sensitization to respond to the antigen's presence. Sensitization begins with...
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Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

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Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
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Ligand Nano-cluster Arrays in a Supported Lipid Bilayer
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Multivalent Tri-Functional T-cell Engagers by Chemically Induced Protein Self-Assembly.

Declan Dahlberg, Brandi McKnight, Abhishek Kulkarni

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    Chemically self-assembled nanorings (CSANs) offer a modular platform for developing T-cell engagers (TCEs). This novel approach enhances TCEs for targeting solid tumors by enabling multispecificity to overcome antigen escape and tumor heterogeneity.

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    Area of Science:

    • Biotechnology
    • Immunology
    • Nanotechnology

    Background:

    • T-cell engagers (TCEs) show promise in cancer immunotherapy but face challenges in solid tumors, including heterogeneity, antigen escape, and limited T-cell infiltration.
    • Developing effective TCEs for solid tumors requires strategies to overcome these barriers and enhance T-cell activity.

    Purpose of the Study:

    • To develop a modular platform using chemically self-assembled nanorings (CSANs) for creating potent multispecific TCEs.
    • To engineer a bifunctional fusion protein and demonstrate its self-assembly into CSANs for targeting EGFR-positive tumor cells and T-cells.
    • To explore the co-assembly of these CSANs with other targeting monomers to create trispecific TCEs for enhanced tumor cell targeting and cytotoxicity.

    Main Methods:

    • Engineered a bifunctional fusion protein (E1-DHFR2-αCD3) with an EGFR-binding domain and an anti-CD3 scFv on a DHFR2 scaffold.
    • Utilized bis-methotrexate to induce self-assembly of monomers into multivalent cis-CSANs.
    • Performed co-culture assays to evaluate binding, internalization, and dose-dependent cytotoxicity of monomers and CSANs against EGFR+ tumor cells and T-cells.
    • Demonstrated co-assembly with additional DHFR2 monomers for creating trispecific CSANs targeting multiple antigens (e.g., GFP, EpCAM).

    Main Results:

    • Both E1-DHFR2-αCD3 monomers and CSANs demonstrated binding to EGFR+ tumor cells and T-cells, followed by internalization.
    • CSANs induced dose-dependent, EGFR- and T-cell-dependent cytotoxicity in co-culture assays, showing reproducibility across T-cell donors.
    • Co-assembly enabled the formation of trispecific CSANs capable of binding multiple antigens and mediating enhanced cytotoxicity.
    • The CSAN platform demonstrated potential for developing multispecific TCEs to address solid tumor challenges.

    Conclusions:

    • The modular CSAN platform provides a versatile strategy for engineering multispecific T-cell engagers.
    • This approach effectively addresses key challenges in solid tumor immunotherapy, such as antigen escape and heterogeneity.
    • Further optimization of antigen combinations within the CSAN platform holds promise for enhancing efficacy across diverse cancer subtypes, including breast cancer.