Dynamics of macrophage trogocytosis of rituximab-coated B cells
| Autor: | Patricia Mero, Theodore Pham, James W. Booth |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Trogocytosis
media_common.quotation_subject Immunology/Innate Immunity lcsh:Medicine Cell Biology/Cell Signaling Flow cytometry Cell Line 03 medical and health sciences Antibodies Monoclonal Murine-Derived Mice 0302 clinical medicine Antigen Cell Biology/Membranes and Sorting Cell Biology/Cytoskeleton medicine Macrophage Animals Cell Biology/Leukocyte Signaling and Gene Expression Internalization Oncology/Hematological Malignancies lcsh:Science Cell Shape 030304 developmental biology media_common 0303 health sciences B-Lymphocytes Multidisciplinary medicine.diagnostic_test biology Macrophages lcsh:R Opsonin Proteins Antigens CD20 Flow Cytometry Molecular biology 3. Good health Cell biology Kinetics Cell culture Immunology/Leukocyte Activation biology.protein Rituximab lcsh:Q Antibody 030215 immunology medicine.drug Research Article |
| Zdroj: | PLoS ONE, Vol 6, Iss 1, p e14498 (2011) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Macrophages can remove antigen from the surface of antibody-coated cells by a process termed trogocytosis. Using live cell microscopy and flow cytometry, we investigated the dynamics of trogocytosis by RAW264.7 macrophages of Ramos B cells opsonized with the anti-CD20 monoclonal antibody rituximab. Spontaneous and reversible formation of uropods was observed on Ramos cells, and these showed a strong enrichment in rituximab binding. RAW-Ramos conjugate interfaces were highly enriched in rituximab, and transfer of rituximab to the RAW cells in submicron-sized puncta occurred shortly after cell contact. Membrane from the target cells was concomitantly transferred along with rituximab to a variable extent. We established a flow cytometry-based approach to follow the kinetics of transfer and internalization of rituximab. Disruption of actin polymerization nearly eliminated transfer, while blocking phosphatidylinositol 3-kinase activity only resulted in a delay in its acquisition. Inhibition of Src family kinase activity both slowed acquisition and reduced the extent of trogocytosis. The effects of inhibiting these kinases are likely due to their role in efficient formation of cell-cell conjugates. Selective pre-treatment of Ramos cells with phenylarsine oxide blocked uropod formation, reduced enrichment of rituximab at cell-cell interfaces, and reduced the efficiency of trogocytic transfer of rituximab. Our findings highlight that dynamic changes in target cell shape and surface distribution of antigen may significantly influence the progression and extent of trogocytosis. Understanding the mechanistic determinants of macrophage trogocytosis will be important for optimal design of antibody therapies. |
| Databáze: | OpenAIRE |
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