Fig. 5.
Fig. 5. Induction of CD27 and analysis of somatic hypermutations in CD27+ B cells derived from naive B cells. / (A) Highly purified CB B cells were stimulated with medium alone, 1 μg/mL anti-CD40 + CD32T, or 0.01% SAC + 50 U/mL IL-2. After 3 days of culture, the cells were stained with anti-CD20–FITC and anti-CD27− biotin followed by streptavidin-PE. (B) Highly purified CB B cells were stimulated with CD40/CD32T. After 3 days, the cells were incubated with anti-CD20–FITC and anti-CD27− biotin followed by streptavidin-PE, and CD27+ and CD27− B cells were sorted by FACStar Plus. The top sequence is the germline gene of VH5 with amino acid translation. Complementarity determining regions are boxed. These data are the representative of 2 different experiments. The same results were obtained when sort-pure adult CD27− B cells were stimulated with CD40/CD32T or SAC + IL-2.

Induction of CD27 and analysis of somatic hypermutations in CD27+ B cells derived from naive B cells.

(A) Highly purified CB B cells were stimulated with medium alone, 1 μg/mL anti-CD40 + CD32T, or 0.01% SAC + 50 U/mL IL-2. After 3 days of culture, the cells were stained with anti-CD20–FITC and anti-CD27− biotin followed by streptavidin-PE. (B) Highly purified CB B cells were stimulated with CD40/CD32T. After 3 days, the cells were incubated with anti-CD20–FITC and anti-CD27− biotin followed by streptavidin-PE, and CD27+ and CD27 B cells were sorted by FACStar Plus. The top sequence is the germline gene of VH5 with amino acid translation. Complementarity determining regions are boxed. These data are the representative of 2 different experiments. The same results were obtained when sort-pure adult CD27 B cells were stimulated with CD40/CD32T or SAC + IL-2.

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