Recommended Information
In vitro–differentiated Th1/Th17/Treg cells
Antibody-Dependent Cell-Mediated Cytotoxicity Assay (ADCC)
Antibody-dependent cellular phagocytosis
Complement-dependent cytotoxicity (CDC)
Cytokine Release Syndrome Risk Assessment (CRS)
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.
Cytokines are predominantly polypeptides, proteins, or glycoproteins with strong antigenicity; immunological techniques can be readily employed to generate antisera, polyclonal antibodies, or monoclonal antibodies against various cytokines. Cytokine detection is of great significance both for basic research in immunology and molecular biology and for elucidating the pathogenic mechanisms of certain diseases and guiding clinical treatment. Immunological assays are among the most commonly used methods for cytokine detection.
CFSE-stained flow cytometry experiment
Carboxyfluorescein diacetate succinimidyl ester (CFSE) staining is a method for detecting cell division. CFSE can readily penetrate the cell membrane; once the intracellular esterases remove the acetate moieties, fluorescence is generated, whereas CFSE that has not entered the cell remains non-fluorescent (Quah et al., 2007).