FIGURE 4 from Overcoming Xenoantigen Immunity to Enable Cellular Tracking and Gene Regulation with Immune-competent “NoGlow” Mice

NoGlow mice reveal the impact of sex and tissue distribution on immune tolerance. A, Male WT (n = 3), CAG Luc-GFP (n = 3), and NoGlow+ (n = 6) or NoGlow− (n = 10) littermates were vaccinated with an Ad encoding WT GFP-Luc and serum was collected after two weeks for anti-Luc or anti-GFP responses. Unvaccinated naïve WT (n = 2) animals were used as baseline controls. B, Luc staining showing protein expression in mammary epithelium of female MMTV Cre NoGlow+ or NoGlow− littermates. C, Luc staining showing protein expression in prostate epithelium of male Pbsn Cre NoGlow+ or NoGlow− littermates. D, Luc staining demonstrating expression in acini and islets in pancreases of male Pdx1 Cre NoGlow+ or NoGlow− littermates. E–G, MMTV Cre (E; n = 4 NoGlow- F, n = 5 NoGlow+ F, n = 2 NoGlow- M, n = 7 NoGlow+ M, n = 3 naïve), Pbsn Cre (F; n = 6 NoGlow- F, n = 6 NoGlow+ F, n = 4 NoGlow- M, n = 8 NoGlow+ M, n = 1 naïve), or Pdx1 Cre (G; n = 2 NoGlow- F, n = 4 NoGlow+ F, n = 4 NoGlow- M, n = 5 NoGlow+ M, n = 2 naïve) littermates were vaccinated with an Ad encoding WT eGFP and Luc. Serum antibody responses to Luc and GFP were determined by ELISA two weeks postvaccination. P values for A, E, F, G are displayed at 1:50 and determined by two-way ANOVA with Tukey correction. H, Normalized ratio of serum anti-Luc or -GFP antibody responses in independent NoGlow+ versus mean of NoGlow− littermates across two independent experiments demonstrating highest tolerance in CMV Cre and Pdx1 Cre animals. P values were determined by one-way ANOVA with Tukey correction. All P values represent mean ± SEM.