Related Experiment Video
Updated: Aug 9, 2026

"Phagosome Closure Assay" to Visualize Phagosome Formation in Three Dimensions Using Total Internal Reflection Fluorescent Microscopy (TIRFM)
Published on: August 26, 2016
Constitutive endocytosis and degradation of CD22 by human B cells
Abstract:
The CD22 B lymphocyte-surface Ag is an important component of the B cell-surface IgM (sIgM)/B cell receptor complex and has been shown to regulate B cell activation. In addition, this molecule has been shown to be an effective target for immunotherapy of B cell malignancies using immunotoxins and radioimmunoconjugates. In this report we describe the internalization and metabolic degradation of this molecule under constitutive conditions and after stimulation of B cells with phorbol dibutyrate or mAbs binding to sIgM, CD19, and CD22. Flow cytometry, "neuraminidase protection," and "neuraminidase shift" assays demonstrated that CD22 is internalized constitutively by unstimulated B cell lines and subsequently degraded in an acidic intracellular compartment (presumably lysosomes) without detectable recycling of the molecule back to the cell surface. Ligation of CD22 with anti-CD22 mAbs markedly increased CD22 internalization but did not affect the rate of intracellular degradation of CD22, suggesting that anti-CD22 mAbs perturb the intracellular trafficking of CD22. In contrast, CD22 internalization and degradation was unaffected by stimulation of B cell lines with phorbol dibutyrate or ligation of other components of the B cell receptor complex (e.g. CD19, sIgM) with mAbs. These patterns of internalization and degradation under constitutive and stimulated conditions contrast with those reported for other lymphoid differentiation Ags (e.g., the TCR, CD3, CD4, and the transferrin receptor), and may help explain the utility of this molecule as a target for immunoconjugate therapy.
More Related Videos
12:48Purification of the Membrane Compartment for Endoplasmic Reticulum-associated Degradation of Exogenous Antigens in Cross-presentation
Published on: August 21, 2017
08:58Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques
Published on: July 5, 2018
Related Concept Videos
Receptor-mediated Endocytosis
Endocytosis
Endocytosis always begins with the plasma membrane enclosing an incoming molecule to form a transport vesicle which, in some cases, can be coated with a protein called ‘clathrin.' Endocytosed material is either sorted through...
Receptor-mediated Endocytosis
Clathrin-Mediated Endocytosis of LDL
One well-characterized example of receptor-mediated endocytosis is the...
Recycling Endosomes and Transcytosis
The recycling endosome is not a single organelle but an extensively tubulated network of recycling pathways. It functions in storing molecules or transporting them across...
Receptor Downregulation in MVBs
The EGFR can initiate signaling pathways that lead to cell proliferation, migration, and differentiation. Overexpression of EGFR stimulates cells to proliferate. Excessive EGFR activation may...
Receptor-Mediated Endocytosis
Clathrin-Mediated Endocytosis of LDL
One well-characterized example of receptor-mediated endocytosis is the...