德晋贵宾厅

Gene targeting strategy for B-hLIGHT mice. The exons 1-4 of mouse Light gene that encode the extracellular protein were replaced by human LIGHT exons 2-5 in B-hLIGHT mice.

Strain specific LIGHT expression analysis in homozygous B-hLIGHT mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLIGHT mice (H/H) stimulated with anti-mCD3ε, anti-mCD28 and mouse IL-2 in vitro for 24h, and analyzed by flow cytometry with species-specific anti-LIGHT antibody. Human LIGHT was exclusively detectable on CD8+T cells in homozygous B-hLIGHT but not in wild-type mice.

Strain specific analysis of LIGHT gene expression in wild-type C57BL/6 mice and B-hLIGHT mice by RT-PCR. Mouse Light mRNA was detectable only in splenocytes of wild-type C57BL/6 mice (+/+). Human LIGHT mRNA was detectable only in homozygous B-hLIGHT mice (H/H), but not in wild-type mice.

Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice and homozygous B-hLIGHT mice (female, 8-week-old, n=3). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Frequencies of T cells, B cells, NK cells, DCs, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hLIGHT mice were similar to those in C57BL/6 mice, demonstrating that humanization of LIGHT does not change the frequency or distribution of these cell types in spleen. Values are expressed as mean ± SEM.

Frequency of leukocyte subpopulations in blood by flow cytometry. Blood cells were isolated from wild-type C57BL/6 mice and homozygous B-hLIGHT mice (female, n=3, 8-week-old). A. Flow cytometry analysis of the blood cells was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, DCs, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hLIGHT mice were similar to those in C57BL/6 mice, demonstrating that humanization of LIGHT does not change the frequency or distribution of these cell types in blood. Values are expressed as mean ± SEM.

Frequency of leukocyte subpopulations in lymph nodes by flow cytometry. Lymph nodes cells were isolated from wild-type C57BL/6 mice and homozygous B-hLIGHT mice (female, n=3, 8-week-old). A. Flow cytometry analysis of the lymph nodes cells was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, CD4+ T cells, CD8+ T cells and Tregs in B-hLIGHT mice were similar to those in C57BL/6 mice. Values are expressed as mean ± SEM.