B-Cd11c-EGFP-DTR-Luc mice have the mouse Itgax (Cd11c) promoter to determine the expression of DTR (Diphtheria toxin receptor), EGFP (enhanced green fluorescent protein) and luciferase.
DCs will be traced by EGFP or Luciferase reporting system in B-Cd11c-EGFP-DTR-Luc mice.
DCs will be depleted in B-Cd11c-EGFP-DTR-Luc mice injected with DT (diphtheria toxin).
The mice can be used to study the origin of mononuclear phagocytes and the specific role of DCs in immune response.
Targeting strategy
Gene targeting strategy for B-Cd11c EGFP-DTR-Luc mice. CD11c co-express EGFP, the DTR for depletion of DCs, and the luciferase for in vivo bioluminescent imaging (BLI) of DCs. EGFP-DTR-Luc expression is evident in CD11c+ cells.
Frequency of leukocyte subpopulations in spleen
Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd11c-EGFP-DTR-Luc mice (Mut/+) (n=3, 6-8-week-old) injected with PBS or DT (50 ng/g body weight) for two consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.0001.
Protein expression analysis
Frequencies of EGFP+ cells and DCs in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd11c-EGFP-DTR-Luc mice (Mut/+) (n=3, 6-8-week-old) injected with PBS or DT (50 ng/g body weight) for two consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of EGFP+ cells and DCs. The frequencies of EGFP+ cells and DCs were decreased in heterozygous mice injected with DT.