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In vitro–differentiated Th1/Th17/Treg cells

CD4+ helper T cells (Th cells) serve as mediators of cellular immunity and play a critical role in activating other immune cells, such as B cells and cytotoxic T cells, as well as in regulating immune responses.

Antibody-Dependent Cell-Mediated Cytotoxicity Assay (ADCC)

Antibodies, as integral components of the immune system, play a crucial role in defending against disease. Antibody-dependent cell-mediated cytotoxicity (ADCC) is one of the mechanisms by which antibodies exert their effector functions: when IgG antibodies specifically bind via their Fab fragments to antigenic epitopes on the surface of target cells—such as virus-infected cells and tumor cells—the Fc portion of the antibody can engage Fc receptors on effector cells, including natural killer (NK) cells, monocytes–macrophages, and neutrophils, thereby triggering the effector cells’ cytotoxic activity and directly killing the target cells. The ability to elicit ADCC against target cells is an important functional criterion for antibody candidates that are directed against cancer-associated antigens.

Antibody-dependent cellular phagocytosis

Antibody-dependent cellular cytotoxicity (ADCC) is one of the mechanisms by which antibody-based therapies exert their antitumor and other therapeutic effects. Currently, therapeutic strategies aimed at enhancing macrophage responses to therapeutic antibodies have garnered significant attention from researchers, including the identification of novel targets and the development of antibodies with enhanced functionality.

Complement-dependent cytotoxicity (CDC)

Complement is a group of heat-labile, enzymatically active proteins found in human and vertebrate serum and tissue fluids, comprising more than 30 soluble and membrane-bound proteins. Complement-dependent cytotoxicity (CDC) refers to the lytic effect on target cells resulting from the formation of a membrane attack complex after complement is activated by specific antibodies that bind to corresponding antigens on the cell membrane via the classical pathway of complement activation. Initially, antibodies bind to complement component C1q, which then triggers the sequential activation of C2 through C9 to form the membrane attack complex, ultimately leading to lysis of the target cell.

Cytokine Release Syndrome Risk Assessment (CRS)

Cytokine release syndrome (CRS) refers to a hyperactive immune response that occurs following infection with pathogenic microorganisms, leading to the rapid activation of numerous immune cells and the massive release of multiple cytokines—including TNF-α, IL-1, IL-6, IL-12, IFN-α, IFN-β, and IFN-γ—within a short period. This results in a severe systemic inflammatory response syndrome. The excessive production of these cytokines can damage tissues and organs, thereby giving rise to a wide range of clinical manifestations. Currently, the standard approach is to closely monitor and target the specific cytokines that trigger the cytokine storm.

Flow Cytometry-Based Cell Characterization Experiments (FACS)

The targets of antibody drugs are primarily disease-associated antigens or specific receptor molecules on the cell surface. Competitive binding between ligands and antibodies is assessed by using flow cytometry to determine the population of antigen-positive cells. By employing antigen-presenting cells in these assays, the spatial conformation of surface antigens more closely resembles their in vivo configuration, thereby yielding results that better reflect physiological conditions.

In vitro–differentiated Th1/Th17/Treg cells


CD4+ helper T cells (Th cells) serve as mediators of cellular immunity and play a critical role in activating other immune cells, such as B cells and cytotoxic T cells, as well as in regulating immune responses.

CD4+ helper T lymphocytes (Th cells) serve as mediators of cellular immunity and play a pivotal role in activating other immune cells, such as B cells and cytotoxic T cells, as well as in regulating immune responses. Undifferentiated CD4+ Th cells are referred to as naïve CD4+ T cells or Th0 precursor cells; upon antigen recognition, these naïve CD4+ T cells undergo clonal expansion and, under the influence of various cytokines, transcription factors, co-stimulatory signals, and adhesion molecules, differentiate into distinct effector cell subsets—namely, the CD4+ Th cell subsets. These subsets include Th1, Th2, Th17, and regulatory T cells (Tregs). Each subset is activated by a specific set of cytokines and transcription factors and exerts unique effector functions by secreting distinct cytokines. Moreover, these cytokines mutually regulate one another, maintaining a delicate and complex immune balance that is essential for optimal immune homeostasis and for the effective execution of cellular immune responses.
Th17 cells represent the third major subset of effector CD4+ T helper cells, following the discovery of Th1 and Th2 cells, and are named for their high-level secretion of IL-17. Th17 cells primarily produce cytokines such as IL-17A, IL-17F, IL-21, and IL-22, and express the transcription factors retinoic acid–related orphan receptor γt (RORγt) and RORα. They play a critical role in host defense against extracellular pathogens, particularly at mucosal and epithelial barriers; however, dysregulation of Th17 cells is associated with the pathogenesis of numerous autoimmune diseases, including multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, psoriasis, and other conditions.
Th17 cells have now become a research hotspot in the field of immunology, with an increasing number of studies exploring these cells and their cytokine products as therapeutic targets for various autoimmune diseases. Consequently, the efficient in vitro generation of large numbers of homogeneous and stable Th17 cells has emerged as a prerequisite for advancing Th17 cell research. However, variations in in vitro differentiation and expansion protocols often result in significant heterogeneity among the derived Th17 cells, characterized by insufficient cell numbers, poor homogeneity, and/or instability, making them prone to transdifferentiation or dedifferentiation.
Therefore, there is an urgent need to develop a rapid, efficient, and robust method for the in vitro differentiation and expansion of Th17 cells, yielding sufficient numbers with high homogeneity and stability. Such a method would be well suited for investigating the molecular mechanisms underlying immune regulation and immune homeostasis, as well as for evaluating the biological activities of therapeutic agents targeting diseases or conditions associated with aberrant Th17 cell activation.

 

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