Figure 3: Survival curve for huNOG-EXL mice across 23 donors (n=5-9 per donor), generated in the Taconic Biosciences humanization core during 2022-2023. All myeloid-supportive HIS mice have limited lifespans due to a range of outcomes including anemia, thrombocytopenia and myeloid cell hyperactivation syndrome. huNOG-EXL has the longest demonstrated lifespan compared to competing models; this varies by donor and can be impacted by environmental and experimental factors. Internal Taconic data.

How Taconic Addresses HIS Mouse Variability

Figure 4: Donor characteristics impact human immune system reconstitution in huNOG-EXL mice, including chimerism levels and frequencies of specific human immune cell types. For huNOG-EXL SA, lots for which the mean falls below Taconic's 25% chimerism threshold are disqualified from sale (lots shown in red). For lots with a mean above the threshold (shown in blue), any individual animals in the lot which score below 25% are disqualified from sale. For huNOG-EXL EA, Taconic selects donors that have previously been validated to reduce the risk of lot failure. Taconic's Field Application Scientists can help you design a successful study plan which appropriately accounts for both inter- and intra-donor variation in HIS mice.

huNOG-EXL EA Mice are Engrafted Using Donor Cells with Proven Engraftment Success

Figure 5: HSCs from the same donor were engrafted twice, on separate days (A and B), to generate two lots of huNOG-EXL mice per donor. The ability of a donor to produce lots where most mice met or failed the QC threshold of ≥25% chimerism was replicated between duplicate lots, with one exception in this test set. To make huNOG-EXL EA mice, Taconic chooses donors with proven engraftment success in order to reduce the risk of lot failure.

Enhanced Myeloid Engraftment in Humanized NOG-EXL Mice

Stable engraftment with significant improvement in myeloid lineage reconstitution and overall cellularity

Figure 6: (a) High humanization rate is observed across multiple donors in huNOG-EXL. NOG-EXL (hGM-CSF/hIL-3 NOG) mice were engrafted with human CD34+ hematopoietic stem cells (HSCs) from n=11 different donors and peripheral blood humanizations was monitored by hCD45 antibody. (b) huNOG-EXL mouse supports development of both myeloid and lymphoid cells.

Better engraftment of myeloid populations in huNOG-EXL mice

Figure 7: Frequencies of CD14⁺ CD16^- (classical) monocytes from FSC-A^hi CD3^- cells (a), HLA-DR⁺ Lin 1^- cells from CD45⁺ cells (b), CD11c^- CD123⁺ plasmacytoid dendritic cells (c), and CD11c⁺ CD123^- myeloid dendritic cells (d), the latter from HLA-DR⁺ Lin 1^- cells, in blood (diamonds) and secondary lymphoid organs (SLO; spleen: circles; axillary lymph node: squares; mesenteric lymph node: triangles) from huNOG-EXL and huNSG mice. NS: Not statistically significant. Adapted from Perdomo-celis, et al. 2019 under Creative Commons Attribution License.

Human immune cell subsets in huNOG-EXL mice

Figure 8: Representative gating strategy from blood cells for the identification of the cell populations evaluated. The number next to the gates represents the respective cell subset found in the adjacent table. Adapted from Perdomo-celis, et al. 2019 under Creative Commons Attribution License.

Tumor Xenograft Models and Immunotherapy Research with huNOG-EXL

Durable anti-tumor response to combination therapy in huNOG-EXL BRCA-deficient tumor model

Figure 9: (a) Tumor growth curve for the MDA-MB-436 NOG-EXL humanized model treated with control or 35 mg/kg niraparib daily for 5 days on and 2 days off for 4 weeks (QD × 5 × 4). (b) Significantly upregulated genes identified with a two-sample t test (p <=0.05, fold change >=1.5) were subjected to enrichment analysis of pathway gene sets and demonstrated a significant enrichment of interferon gamma signature and interferon alpha signature genes in niraparib-treated samples. (c-e) Niraparib promoted tumor immune cell infiltration in BRCA-deficient MDA-MB-436 huNOG-EXL humanized tumor model. (f) Tumor growth in the BRCA-deficient MDA-MB-436 model in huNOG-EXL mice treated with 200 mg anti-PD-1 (pembrolizumab) on days 0, 4, 9, 13, 18, 22, and 28; 35 mg/kg niraparib daily for 5 days on and 2 days off for 4 weeks; and the combination of these agents. Adapted from Wang, et al. 2019 under Creative Commons Attribution License.

Immunophenotyping and anti-CTLA4 human immune response in huNOG-EXL mice

Figure 10: (a) Anti-CTLA4 antibody pharmacokinetics in huNOG-EXL mice. (b) Splenic immunophenotyping of huNOG-EXL mice treated with anti-CLTA4 antibodies. NF: nonfucosylated. Data provided by Dr. Kathryn Fraser, Takeda.

Infectious Disease Research with huNOG-EXL

HIV infection in huNOG-EXL mice affects the frequencies of human immune cells

Figure 11: Frequencies of CD3+ cells (a), CD20+ cells (b), CXCR5+ CD4+ T-cells (c), CXCR5+ CD8+ T-cells (d), CD14+ CD16- (classical) monocytes from FSC-Ahi CD3- cells (e), HLA-DR+ Lin 1- cells from CD45+ cells (f), CD11c- CD123+ plasmacytoid dendritic cells from HLA-DR+ Lin 1- cells (g), CD56dim (h), and CD56bright NK cells (i) in huNOG-EXL mice after infection with HIV. Adapted from Perdomo-celis, et al. 2019 under Creative Commons Attribution License.