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A monoclonal antibody specific for

2016 
Dendritic cells (DC) are a small subpopulation of lymphoid cells with distinctive cytologic features, surface prop- erties, and functions. This report describes the,DC-specific an- tibody (Ab) secreted by clone 33D1. Rat spleen cells immune to mouse DC were fused to the P3U myeloma. Hybrid culture su- pernatants were screened simultaneously against DC, a macro- phage (M4>) 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' M4> (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: MO from spleen, peritoneal cavity, and blood; three MO cell lines; lymphocytes; granulocytes; plate- lets; and erythroid cells. DC continued to express the 33D1 antigen after 4 days in culture, whereas'M1 and lymphocytes did not ac- quire 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.
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