hIL12B/hIL23A/hTNFa-Tg

Table.1 Blood biochemistry examination in 6-week-old HO hTNFa-Tg/hIL23A/hIL12B mice (n=3 /group).

Table.2 Blood routine examination in 6-week-old HO hTNFa-Tg/hIL23A/hIL12B mice (n=2/group).

Fig.1 Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from WT C57BL/6 (WT) mice and HO hTNFa-Tg/hIL23A/hIL12B (HO) mice (male, 6-week-old, n=3). Flow cytometry analysis of frequencies of lymphocyte and myeloid subpopulations in the spleen. Percentages of T cells, B cells, and NK cells in the spleen of HO hTNFa-Tg/hIL23A/hIL12B mice were similar to those in C57BL/6 mice.  However, there is a trend of higher frequency of CD11b+myeloid cells, DCs, neutrophils, monocytes, and macrophages in the spleen of humanized mice than in WT C57BL/6 mice, which might be related to the overexpression of humanized TNFα in the mice. Values are expressed as mean ± SEM.

Fig.2 Frequency of leukocyte subpopulations in lymph notes by flow cytometry. Lymph notes cells were isolated from WT C57BL/6 (WT) mice and hTNFa-Tg/hIL23A/hIL12B (HO) mice (male, 6-week-old, n=3). Flow cytometry analysis of frequencies of lymphocyte and myeloid subpopulations in the lymph notes. Percentages of T cells, B cells, NK cells, CD11b+myeloid cells, DCs, neutrophils, monocytes, and macrophages in lymph notes of HO hTNFa-Tg/hIL23A/hIL12B mice were similar to those in C57BL/6 mice, demonstrating that humanization of these genes does not change the frequency or distribution of these cell types in lymph notes. Values are expressed as mean ± SEM.

Fig.3 Human TNF-α levels in serum of humanized hTNFa-Tg/hIL23A/hIL12B mice following LPS stimulation. Wild-type C57BL/6 mice and humanized hTNFa-Tg/hIL23A/hIL12B mice(male, 6 weeks old) were challenged with a single intraperitoneal injection of LPS. Serum concentrations of human TNF-α were measured by ELISA 2 hours post-injection.  Human TNF-α was highly expressed in homozygous humanized  hTNFa-Tg/hIL23A/hIL12B mice. Data are presented as mean ± SEM from three male mice per group.