B-Rag2/IL2rg KO rats

Basic Information

Description

The recombinant-activating gene 2 (Rag2) plays an important role in the rearrangement and recombination of the genes of immunoglobulin and T cell receptor molecules during the initiation of V(D)J recombination. Loss of Rag2 protein leads to no mature T and B cells, the critical components of the adaptive immune system.

Il2rg is known as the interleukin receptor common gamma chain, which is an important signaling component of many interleukin receptors, including those of interleukin -2, interleukin -4, interleukin -7 and interleukin -21. Mutations in this gene cause X-linked severe combined immunodeficiency. 

This rat can be used to support research in many areas including immune system defections, virology research, inflammation research, cancer research, xenograft/transplant host etc.

Targeting Strategy

Gene targeting strategy for B-Rag2/Il2rg KO rats. Part of the sequences of exon 3 in rat Rag2 gene and part of the sequences of exon 2 in rat Il2rg gene were deleted in B-Rag2/Il2rg KO rats. The background strain is SD rats.

B-Rag2/Il2rg KO rats exhibit the smallest thymus size

The thymus size of B-Rag2/Il2rg KO rats was the smallest compared to that of wild-type SD rats and B-Rag2 KO rats. Gross anatomy of thymuses in male and female wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats were showed (n=3, 7-week-old). Thymus size of B-Rag2 KO rats were significantly reduced compared to that of wild-type SD rats. But the thymus size of B-Rag2/Il2rg KO rats were further reduced compared to B-Rag2 KO rats. 

B-Rag2/Il2rg KO rats exhibit the significantly reduced thymus and spleen weight

The size and weight of thymus and spleen of B-Rag2/Il2rg KO rats were significantly reduced compared to that of wild-type SD rats. Thymus and spleen were isolated from male and female wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats and weighed (n=3, 7-week-old). Thymus size and weight of B-Rag2 KO rats were significantly reduced compared to that of wild-type SD rats. But the thymus size and weight of B-Rag2/Il2rg KO rats were further reduced compared to B-Rag2 KO rats. Spleen size and weight of B-Rag2 KO rats and B-Rag2/Il2rg KO rats were significantly reduced compared to that of wild-type SD rats.

Analysis of leukocyte subpopulation in thymus

Complete loss of T cells in thymus of homozygous B-Rag2 KO rats and B-Rag2/Il2rg KO rats. Thymuses were collected from male and female wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats (n=3, 7-week-old). Leukocyte subpopulations were analyzed by flow cytometry analysis. A. Representative FACS plots. B. Statistical analysis of FACS. Results showed that total T cells, CD4+ T cells and CD8+ T cells were only detectable in thymuses of wild-type SD rats. But none of the T cells were detectable in thymuses of B-Rag2 KO rats or B-Rag2/Il2rg KO rats. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in spleen of male rats

Complete loss of T cells, B cells and NK cells in spleen of homozygous male B-Rag2/Il2rg KO rats. Spleens were collected from male wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats (n=3, 7-week-old). Leukocyte subpopulations were analyzed by flow cytometry analysis. Results showed that T cells and B cells were not detectable in spleen of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in spleen of female rats

Complete loss of T cells, B cells and NK cells in spleen of homozygous female B-Rag2/Il2rg KO rats. Spleens were collected from female wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats (n=3, 7-week-old). Leukocyte subpopulations were analyzed by flow cytometry analysis. Results showed that T cells and B cells were not detectable in spleen of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in spleen

Complete loss of T cells, B cells and NK cells in spleen of homozygous male and female B-Rag2/Il2rg KO rats. Spleens were collected from male and female wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats (n=3, 7-week-old). Leukocyte subpopulations were analyzed by flow cytometry analysis. Results showed that T cells and B cells were not detectable in spleen of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in blood of male rats

Complete loss of T cells, B cells and NK cells in blood of homozygous male B-Rag2/Il2rg KO rats. Blood cells were collected from male wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats (n=3, 7-week-old). Leukocyte subpopulations were analyzed by flow cytometry analysis. Results showed that T cells and B cells were not detectable in spleen of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in blood of female rats

Complete loss of T cells, B cells and NK cells in blood of homozygous female B-Rag2/Il2rg KO rats. Blood cells were collected from female wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats (n=3, 7-week-old). Leukocyte subpopulations were analyzed by flow cytometry analysis. Results showed that T cells and B cells were not detectable in spleen of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in blood

Complete loss of T cells, B cells and NK cells in blood of homozygous male and female B-Rag2/Il2rg KO rats. Blood cells were collected from male and female wild-type SD rats, B-Rag2 KO rats and B-Rag2/Il2rg KO rats (n=3, 7-week-old). Leukocyte subpopulations were analyzed by flow cytometry analysis. A. Representative FACS plots. B. Statistical analysis of FACS. Results showed that T cells and B cells were not detectable in spleen of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats. Values are expressed as mean ± SEM.

MV-4-11 human leukemia xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of MV-4-11 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human biphenotypic B myelomonocytic leukemia cell line MV-4-11 (2×107) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, MV-4-11 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. B-Rag2/Il2rg KO rats can be used for efficacy studies.

THP-1 human leukemia xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of THP-1 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human acute monocytic leukemia cell line THP-1 (2×107) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, THP-1 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats, and the two strains of rats can be used for efficacy studies.  

MIA PaCa-2 human pancreas cancer xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of MIA PaCa-2 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human pancreas cancer cell line MIA PaCa-2 (2×107) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, MIA PaCa-2 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats, and the two strains of rats can be used for efficacy studies.  

NCI-H2122 human lung cancer xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of NCI-H2122 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human non-small cell lung cancer cell line NCI-H2122 (2×107) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, NCI-H2122 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats, and the two strains of rats can be used for efficacy studies.

NCI-H1373 human lung cancer xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of NCI-H1373 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human lung cancer cell line NCI-H1373 (2×107) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, NCI-H1373 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats, and the two strains of rats can be used for efficacy studies.  

A549 human lung cancer xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of A549 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human lung cancer cell line A549 (2×107) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, A549 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats, and the two strains of rats can be used for efficacy studies.  

RT-112 human bladder cancer xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of RT-112 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human bladder cancer cell line RT-112 (1×107) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, RT-112 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats, and the two strains of rats can be used for efficacy studies.  

HCC1954 human breast cancer xenograft model in B-Rag2/Il2rg KO rats and B-Rag2 KO rats

Subcutaneous homograft tumor growth of HCC1954 cells in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. Human breast cancer cell line HCC1954 (25×106) were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats and B-Rag2 KO rats (n=5). (A)Tumor volume. (B) Body weight change. As shown in panel A, HCC1954 cells were able to establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats, and the two strains of rats can be used for efficacy studies.  

NCI-H460 human large cell lung cancer xenograft model in B-Rag2/Il2rg KO rats

Subcutaneous homograft tumor growth of NCI-H460 cells in B-Rag2/Il2rg KO rats. Human large cell lung cancer cell line NCI-H460 were mixed with Matrigel and inoculated subcutaneously into B-Rag2/Il2rg KO rats (n=6). (A)Tumor volume. (B) Body weight change. As shown in panel A, NCI-H460 cells were able to establish tumors in B-Rag2/Il2rg KO rats, and this strain of rat can be used for efficacy studies. 

In vivo efficacy of cisplatin

Antitumor activity of cisplatin in B-Rag2/Il2rg KO rats. (A) Cisplatin inhibited HCC1954 tumor growth in B-Rag2/Il2rg KO rats. Human breast invasive ductal carcinoma cells (2E7) were subcutaneously implanted into B-Rag2/Il2rg KO rats (female, 9-week-old, n=6). The rats were grouped when tumor volume reached approximately 300-400 mm3, at which time they were treated with cisplatin with doses indicated in panel. (B) Body weight changes during treatment. As shown in panel A, cisplatin was efficacious in controlling tumor growth in B-Rag2/Il2rg KO rats, demonstrating that B-Rag2/Il2rg KO rats provide a powerful preclinical model for in vivo evaluation of chemotherapies. Values are expressed as mean ± SEM.

Summary

Gross anatomy of thymus and weight of thymus and spleen:

• Thymus size and weight of B-Rag2 KO rats were significantly reduced compared to that of wild-type SD rats. But the thymus size and weight of B-Rag2/Il2rg KO rats were further reduced compared to B-Rag2 KO rats. Spleen size and weight of B-Rag2 KO rats and B-Rag2/Il2rg KO rats were significantly reduced compared to that of wild-type SD rats. 

 

Leukocyte subpopulation analysis：

• Thymus: Total T cells, CD4+ T cells and CD8+ T cells were only detectable in thymuses of wild-type SD rats. But none of the T cells were detectable in thymuses of B-Rag2 KO rats or B-Rag2/Il2rg KO rats. 

• Spleen: T cells and B cells were not detectable in spleen of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats.

• Blood: T cells and B cells were not detectable in blood of B-Rag2 KO rats and B-Rag2/Il2rg KO rats. NK cells were also not detectable in B-Rag2/Il2rg KO rats. But the percentage of NK cells in B-Rag2 KO rats were relatively higher than that in wild-type SD rats. The percentages of DCs, CD11b+CD43+ monocytes, CD11b+CD68+ macrophages, CD11b+CD43+CD68+ monocytes/macrophages (Mo/Mφ) and neutrophils in B-Rag2 KO rats and B-Rag2/Il2rg KO rats were relatively higher than that in wild-type SD rats. 

Cell line derived xenograft model (CDX) can successfully constructed in B-Rag2/Il2rg KO rats and B-Rag2 KO rats: 

Human tumor cell lines can successfully establish tumors in B-Rag2/Il2rg KO rats and B-Rag2 KO rats. The cell lines which have been verified are MV-4-11, THP-1, MIA PaCa-2, NCI-H2122, H460, NCI-H1373, A549, RT-112, HCC1954.

In vivo efficacious：

Chemotherapy drug cisplatin was efficacious in controlling tumor growth in B-Rag2/Il2rg KO rats, demonstrating that B-Rag2/Il2rg KO rats provide a powerful preclinical model for in vivo evaluation of chemotherapies.