C-NKG

Next-generation immunodeficient mouse models such as C-NKG mice have several features that are advantageous for cancer immunotherapy studies:

● Lacks mature T, B, and NK cells

● Decreased complement activity

● Dysfunction of macrophages and dendritic cells

● Extremely low incidence of T and B cell leakage with age

● Extremely low incidence of lymphoma (different from NOD SCID model)

● Can be used in both long-term and short-term experiments

● Does not develop diabetes

● C-NKG mice have prolonged life spans compared with the NOD SCID mice, which have a high tendency of developing thymic lymphoma.

● C-NKG mice demonstrate better CDX/PDX engraftment rates compared to NOD SCID mice.

● High efficiency in transplantation of human hematopoietic stem cells (HSCs), peripheral blood mononuclear cells (PBMCs), patient-derived xenografts (PDXs) or adult stem cells and tissues.

● May be used for research on tumors, immunity, autoimmune diseases, immunotherapy vaccines, GvHD/transplantation, safety evaluations, and more.

1. Prkdc gene mutation

Figure 1. Prkdc^scid mutation is produced by TAT → TAA in exon 84 of Prkdc gene.
The sequencing result shows that C-NKG mice carry Prkdc^scid mutation.

2. Il2rg gene knockout

Figure 2. The II2rg gene of NKG mouse was detected by PCR and the result shows that the II2rg gene of NKG mouse was successfully knocked out.
The band size of wild type was 1430 bp and that of the knockout fragment was 388 bp.

3. Detection of B, T, and NK cells in peripheral blood of C-NKG mouse

                       
Figure 3. The peripheral blood of C-NKG mouse, a severe combined immunodeficiency (SCID) mouse model, is severely deficient in B, T, and NK cells.
Peripheral blood samples of BALB/c and C-NKG mice were collected, followed by the performance of flow cytometric immunophenotypic analysis and statistical comparison of the composition of T, B and NK cells. The results show that B cells (CD3^-CD19⁺), T cells (CD3⁺CD19^-), helper T cells (CD3⁺CD4⁺CD8^-), cytotoxic T cells (CD3⁺CD4^-CD8⁺) and NK cells (CD335⁺CD3^-) in the peripheral blood of C-NKG mice were almost completely absent while comparing with BALB/c mice.

4. Detection of B, T, and NK cells in lymphoid tissue of C-NKG mice

Figure 4. The lymphoid tissue of C-NKG mouse, a severe combined immunodeficiency (SCID) mouse model, is severely deficient in B, T and NK cells.
Lymphoid tissue samples of BALB/cand C-NKGmice were collected, followed by flow cytometric immunophenotypic analysis and statistical comparison of the composition of T, B and NK cells. The results show that B cells (CD3^-CD19⁺), T cells (CD3⁺CD19^-), helper T cells (CD3⁺CD4⁺CD8^{-), cytotoxic T cells (CD3+CD4-CD8+) and NK cells (CD335+CD3-) in the lymphoid tissue of C-NKGmice were almost completely absent, ascompared with BALB/cmice.}

5. huPBMC-C-NKG

Figure 1. Survival curve of huPBMC-C-NKG mice.

The huPBMC-C-NKG mice gradually died with the onset of graft-versus-host disease (GvHD).

Figure 2. Changes in the content of human CD45+ cells in peripheral blood after the transplantation of human PBMCs into C-NKG mice.

After PBMC transplantation, the content of human leukocytes in the peripheral blood of C-NKG mice gradually increased, and the average proportion of CD45+ was about 50% after 3 weeks.

Figure 3. Changes in the content of human CD3+ cells in peripheral blood after transplantation of human PBMCs into C-NKG mice.

The reconstitution of huPBMC-C-NKG mice was dominated by T cells.

6. Human tumor cell line xenograft (CDX) model

Figure 1. Tumor growth curve of human hepatoma cell Huh7 subcutaneous xenograft

Figure 2. Tumor growth curve of human gastric cancer cell HGC-27 subcutaneous xenograft.

Figure 3. Human colon adenocarcinoma SW620 subcutaneous xenograft tumor growth curve

Cells were subcutaneously inoculated into both C-NKG and NOD/Scid mice, and tumor volume was measured at different time points. The cell seeding amount was 5×10⁶/cell and the data were presented in the form of Mean±SEM. The results showed that human liver cancer cells Huh7, human gastric cancer cells HGC-27 and human colon adenocarcinoma cells SW620 could effectively establish tumor models in C-NKG mice.