Figure 3
Figure 3. Generation of hu-BLT mice expressing b12-IgA through HSPC gene transfer. (A) A schematic diagram of the generation of hu-BLT mice transduced with human b12-IgA gene (hu-BLT-b12a mice). (B) Peripheral and mucosal reconstitution of a human immune system in hu-BLT-b12a mice. Flow cytometry shows human cell engraftment in various tissues isolated at 14 to 16 weeks after transplantation; PBMC indicates peripheral blood mononuclear cells; SPL, spleen; BM, bone marrow; Gut IEL, intestinal intraepithelial lymphocytes; Gut LPL, intestinal lamina propria lymphocytes; and genital, genital tract lymphocytes. (C) Percent engraftment of human B and T lymphocytes in periphery (Ctrl-transduced mice n = 7, b12-IgA-transduced mice n = 21) and the secretion of human IgA in the plasma (Ctrl-transduced mice, n = 5, b12-IgA-transduced mice, n = 12) of hu-BLT mice either transduced with b12-IgA gene or the control gene (mean ± SEM). (D) Immunohistochemical staining for human CD3 in spleen and small intestine tissues and human IgA-Producing cells in spleen and female reproductive tract (FRT) of hu-BLT-b12a mice. Samples were examined on an Olympus BX-51 microscope (20× objective lens) and photographed using a Spot Digital Camera. (E) Bioluminescence images of hu-BLT mice transduced with IgL chain promoter-driven luciferase transgene. The representative images of live animals displayed the distribution of human B-lymphocytes derived from transplanted human HSPCs that express transgenes; (Ei) ventral view of reference, (Eii) dorsal view of reference, and (Eiii) lateral view of reference. Strong bioluminescent signals from mucosal associated lymphoid tissues near gut, lung and genital tract area are seen in ventral view (Ei). Secondary lymphoid tissue (spleen) signals are seen in dorsal (Eii) and lateral view (Eiii). A luciferin-injected mouse was sacrificed after 5 minutes of incubation and dissected immediately to excise organs and tissues. The representative images of GI tract (Eiv), spleen (Ev), and genital tract (Evi) show tissue specificity of transgene expression. (F) Luminescent signals from hu-BLT mice described above at 18 weeks after transplantation. Signals recorded in photons/second were acquired from each indicated areas in the whole animal image.

Generation of hu-BLT mice expressing b12-IgA through HSPC gene transfer. (A) A schematic diagram of the generation of hu-BLT mice transduced with human b12-IgA gene (hu-BLT-b12a mice). (B) Peripheral and mucosal reconstitution of a human immune system in hu-BLT-b12a mice. Flow cytometry shows human cell engraftment in various tissues isolated at 14 to 16 weeks after transplantation; PBMC indicates peripheral blood mononuclear cells; SPL, spleen; BM, bone marrow; Gut IEL, intestinal intraepithelial lymphocytes; Gut LPL, intestinal lamina propria lymphocytes; and genital, genital tract lymphocytes. (C) Percent engraftment of human B and T lymphocytes in periphery (Ctrl-transduced mice n = 7, b12-IgA-transduced mice n = 21) and the secretion of human IgA in the plasma (Ctrl-transduced mice, n = 5, b12-IgA-transduced mice, n = 12) of hu-BLT mice either transduced with b12-IgA gene or the control gene (mean ± SEM). (D) Immunohistochemical staining for human CD3 in spleen and small intestine tissues and human IgA-Producing cells in spleen and female reproductive tract (FRT) of hu-BLT-b12a mice. Samples were examined on an Olympus BX-51 microscope (20× objective lens) and photographed using a Spot Digital Camera. (E) Bioluminescence images of hu-BLT mice transduced with IgL chain promoter-driven luciferase transgene. The representative images of live animals displayed the distribution of human B-lymphocytes derived from transplanted human HSPCs that express transgenes; (Ei) ventral view of reference, (Eii) dorsal view of reference, and (Eiii) lateral view of reference. Strong bioluminescent signals from mucosal associated lymphoid tissues near gut, lung and genital tract area are seen in ventral view (Ei). Secondary lymphoid tissue (spleen) signals are seen in dorsal (Eii) and lateral view (Eiii). A luciferin-injected mouse was sacrificed after 5 minutes of incubation and dissected immediately to excise organs and tissues. The representative images of GI tract (Eiv), spleen (Ev), and genital tract (Evi) show tissue specificity of transgene expression. (F) Luminescent signals from hu-BLT mice described above at 18 weeks after transplantation. Signals recorded in photons/second were acquired from each indicated areas in the whole animal image.

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