Langerhans cell, lymph node, lymphatic system, monoclonal antibody
Dendritic cells (DC) are a small subpopulation of lymphoid cells with distinctive cytologic features, surface properties, and functions. This report describes the DC-specific antibody (Ab) secreted by clone 33D1. Rat spleen cells immune to mouse DC were fused to the P3U myeloma. Hybrid culture supernatants were screened simultaneously against DC, a macrophage (MΦ) cell line, and mitogen-stimulated lymphoblasts. 33D1 Ab specifically killed 80-90% of DC from spleen and lymph node, but no other leukocytes, including Ia+ and Ia- MΦ (Ia, I-region-associated antigen). Quantitative binding studies with 3H-labeled 33D1 Ab showed that DC had an average of 14,000 binding sites per cell. Binding to DC was inhibited with Fab fragment of 33D1 Ab but not with a panel of other monoclonal antibodies, including anti-Ia Ab. Adherence and flotation procedures that enriched for DC enriched for 3H-labeled 33D1 Ab binding in parallel. 33D1 antigen was not detectable on: MΦ from spleen, peritoneal cavity, and blood; three MΦ cell lines; lymphocytes; granulocytes; platelets; and erythroid cells. DC continued to express the 33D1 antigen after 4 days in culture, whereas MΦ and lymphocytes did not acquire it. Quantitative and autoradiographic studies confirmed that spleen and lymph node suspensions contain less than 1% DC. We conclude that 33D1 Ab detects a stable and specific DC antigen and can be used to monitor DC content in complex lymphoid mixtures.
Nussenzweig, M. C., R. M. Steinman, M. D. Witmer, and B. Gutchinov. 1982. "A Monoclonal Antibody Specific for Mouse Dendritic Cells." Proceedings of the National Academy of Sciences of the United States of America 79 (1): 161-165