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To generate nondeleting mice (3-83 ND mice), we backcrossed 3-83 transgenic mice (13), which express IgM and IgD forms of the 3-83 antibody, a minimum of 10 times onto a B10,D2nSn/J background.
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b double Tg mice presented a peripheral deletion phonotype similar to those described using 3-83 mice crossed to mice expressing Kb in the liver under the control of the sheep metallothionein promoter (13).
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3. The following antibody reagents were used: mouse anti-IgM conjugated with fluorescein isothiocyanate (FITC) (Zymed), mouse anti-B220 conjugated with phycoerythrin (PE) (Caltag), mouse anti-CD8 conjugated with FITC, and mouse anti-CD4 conjugated with PE (Pharmingen). Flow cytometric analysis was done with a FacScan (Becton Dickinson) and the data were analyzed by Cell Quest software.
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39
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note
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Spleens were flash-frozen in O.C.T. embedding medium (Miles), and 6-μm sections were cut and fixed in acetone. Serial sections were blocked for 10 min with PBS and 10% fetal calf serum, incubated with 2.4G2 culture supernatant for 30 min, then stained for 30 min with PE-labeled anti-B220 and FITC-labeled IgM, idiotype, or isotype control monoclonal antibodies.
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b mice.
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