SMOC™ Mouse Models For Immunology Research Mouse Models For Immune Cell Tracking
Reporter gene-based molecular imaging is a powerful means to detect the location and function of diverse cell populations in vivo. Generation of immune cells marked with imaging reporter genes permits tracking of immune responses to pathogens and cancer in experimental systems.
SMOC has established a list of mouse models in which distinct immune cell types were labeled by luciferase and EGFP, providing simpler, highly sensitive, robust means for immune cell infiltrate and in vivo drug efficacy studies.
Cd8a-Luc-EGFP mice luc-EGFP ●● Cd8a mutant mice express knocked-in luciferase Strain Name Cat. NO. and EGFP genes from the Cd8a locus--without disrupting expression of the endogenous Cd8a gene. Cd8a-Luc-EGFP NM-KI-18030 When cell populations are activated, identify Cd8a transcriptional activity both in vivo and in vitro. ●● These mice may be useful for evaluate drug anti- tumor activity.
Cd8a WT No treatment PD1 antibody treatment 1day 2.0 6.0
5.0 1.5 WT Tluc 4.0
C57-3 BLOOD1: P3 CD8A 112#HE1 BLOOD1:P3 6
6 6 6 10 Q2-UL(46.50%) Q2-UR(0.00%) 10 Q2-UL(23.69%) Q2-UR(13.17%)
1.0 ×10 3.0 ×10 5 5 10 10 2.0 4 0.5 4 10 10 APC-A APC-A 1.0 3 10 3 10 Radiance Radiance 2 2 0 0 p/sec/cm /sr p/sec/cm /sr Q2-LL(53.44%) Q2-LR(0.07%) Q2-LL(63.06%) Q2-LR(0.08%)
CD8a Color Scale 2 3 4 5 6 2 3 4 5 6 Min = 4.00e4 10 10 10 10 10 10 10 10 10 10 Max = 2.00e6 FITC-A FITC-A EGFP No stimulation MC38 tumor model
Figure 1. Expression of EGFP in PBMC-Derived CD8+ T cells of Cd8a-Luc- Figure 2. In the absence of any stimulation, the in vivo imaging system luc-EGFP EGFP mice is detected by FACS. detected that Cd8a mice showed significant fluorescence.
www.modelorg.com 2 SMOC Mouse Models For Immunology Research 1 day 10 day 17 day MC38
2.0 B16F10
1.5 7 ×10 1.0
Figure 3. Bioluminescence imaging of CD8+ T cells in Cd8a- Luc-EGFP mice after inoculation tumor cells. Enhanced T 0.5 cell recruitment and infiltration into the tumor mass was B16F10.SIY observed on Cd8a-Luc-EGFP mice following the implantation
Radiance of MC38 colon cancer cells and antigenic SIY-expressing B16 2 p/sec/cm /sr melanoma cells (but not the regular B16 cells).
No treatment PD1 antibody treatment 1day 6.0
5.0
4.0 6 ×10 3.0
2.0
1.0 Figure 4. In vivo imaging validation of MC38 tumor-bearing luc-EGFP Radiance Cd8a mouse model. After 1 day of stimulation with PD1 2 p/sec/cm /sr antibody, the fluorescence was significantly enhanced. MC38 tumor model
More mouse models
Strain Name Cat.No. Target cells
Cd19-EGFP-Luc NM-KI-200058 B cells
+ Cd8a-Luc-EGFP NM-KI-18030 Cd8 T cells
Foxp3-Luc-tdTomato NM-KI-18034 Regulatory T lymphocytes
Itgax-tdTomato-Luc NM-KI-190116 Dendritic cells
Lyz2-mtagBFP-Luc NM-KI-200029 Myeloid cell lineage (monocytes, mature macrophages and granulocytes)
Ncr1-mtagBFP-Luc NM-KI-200030 Natural killer cells
SMOC Mouse Models For Immunology Research 3 www.modelorg.com Mouse Models For Immune Cell Ablation
The immune system is an organization of cells and molecules with specialized roles in the different immune responses. To decipher the functional contributions of specialized cell populations in the immune response, SMOC has established a list of DTREGFP mouse models in which distinct immune cell types can be ablated. These DTREGFP mice provide a diphtheria toxin inducible system that administration of diphtheria toxin results in depletion of specific cell populations. Cell ablation is a in vivo valuable complement to mutagenesis for experimentally defining specific cell functions in physiology and pathophysiology.
Cd8a-DTREGFP Mice DTREGFP ●● Cd8a mutant mice express knocked-in human Strain Name Cat. NO. diphtheria toxin receptor and EGFP genes from the Cd8a locus--without disrupting expression of the Cd8a-DTREGFP NM-KI-190045 endogenous Cd8a gene. + ●● These mice may be useful for CD8 T cells visualizing and ablation.
8JMEUZQF )FUFSP[ZHPVT $$%B%53&('11 $%B%53&('11 26- 263 26- 263
" "
$ $ '*5 '*5 &('1
2-- 2-3 2-- 2-3 DTREGFP/+ Figure 5. EGFP expression in PBMC of CD8a mice and C57BL/6 mice was detected by FACS. (In collaboration with CrownBio) $%B"1$
www.modelorg.com 4 SMOC Mouse Models For Immunology Research A 1SFEPTF %BZ %BZ %BZ <$% >$%$% <$% >$%$% <$% >$%$% <$% >$%$%
$% $% $% $%
$% $% $%
$% $% $% $% $% $#-
$% $% $% $%
<$% >$%$% <$% >$%$% <$% >$%$% <$% >$%$%
$% $% $% $%
53('1 $% $% % $%
$% $% $% $% $%
$%B*3&4
$% $% $% $%
B Change of % CD8 over time
CD8a-IRES-DTR/EGFP (N=2) 50 Naïve control mice (N=8)
38.7 40 37.3 36.1 33.6 32.4 30
20 CD8 % in CD3 + Figure 6. FACS analysis of the percent of CD8 T cell in peripheral blood Day 1,3,5: DT dose administration DTREGFP/+ 10 of MC38 tumor-bearing CD8a mice and C57BL/6 mice (A and B). Nearly complete depletion was achieved only after 1 dose administration 0.7 0.1 0.8 0 of DT and the depletion effect lasted to day 14. (In collaboration with Pre-dose Day 3 Day 7 Day 14 CrownBio) Days
CD8a-IRES-DTR/EGFP 3000 Naïve control mice (N=8) ) 3
2500
2000
1500
1000 Day1,3,5: DT dose administration DTREGFP/+ Figure 7. The growth of MC38 tumors in CD8a mice and C57BL/6 500 DTREGFP/+ Tumor Volume±SEM (mm mice. CD8a mice and C57BL/6 mice were inoculated with MC38 3 DTREGFP/+ 0 colon cancer cells. After the tumors grew to 100 mm , the CD8a 0 2 4 6 8 10 12 14 16 18 20 22 24 mice were injected with diphtheria toxin. (In collaboration with CrownBio) Days
SMOC Mouse Models For Immunology Research 5 www.modelorg.com Ncr1-DTREGFP Mice
●● NCR1 is expressed specifically on natural killer (NK) Strain Name Cat. NO. cells and possibly on related innate lymphocytes. DTREGFP Ncr1-DTREGFP NM-KI-190044 Ncr1 mutant mice express knocked-in human diphtheria toxin receptor and EGFP genes from the Ncr1 locus—without disrupting expression of the endogenous Ncr1 gene. ●● These mice may be useful for natural killer cells visualizing and ablation.
A Predose Day3 Day7 Day14 Day21 [LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335
5 5 5 5 5 10 NK: 2.02% NKT: 0.39% 10 NK: 2.12% NKT: 0.87% 10 NK: 1.85% NKT: 0.65% 10 NK: 1.65% NKT: 0.67% 10 NK: 2.00% NKT: 1.01%
4 4 4 4 4 10 10 10 10 10 CD335 CD335 CD335 3 CD335 CD335 3 10 3 3 3 C57BL/6 10 10 10 10 0 0 0 0 0 3 3 3 3 ‒10 3 ‒10 ‒10 ‒10 ‒10 4 5 4 5 3 4 5 4 5 4 5 0 10 10 0 10 10 0 10 10 10 0 10 10 0 10 10 CD3 CD3 CD3 CD3 CD3
[LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335 [LY6G-LY6C-] CD3/CD335
5 5 5 5 5 10 NK: 2.03% NKT: 0.30% 10 NK: 0.40% NKT: 0.30% 10 NK: 0.46% NKT: 0.68% 10 NK: 0.91% NKT: 0.46% 10 NK: 1.78% NKT: 0.70%
4 4 4 4 4 10 10 10 10 10 CD335 CD335 3 CD335 3 3 CD335 3 CD335 3 10 10 10 10 10 0 0 0 0 0 3 3 3 3 ‒10 3 ‒10 ‒10 ‒10 ‒10 Ncr1-IRES-DTRGFP 4 5 4 5 3 4 5 4 5 4 5 0 10 10 0 10 10 0 10 10 10 0 10 10 0 10 10 CD3 CD3 CD3 CD3 CD3
B Change of % NK over time
Ncr1-IRES-DTR/EGFP(N=2) 3.0 Naïve control mice(N=8)
2.5 2.1 2.1 2.0 1.9 2.0 2.0 1.6 1.5 1.8 NK % in Gated 1.0 0.4 0.5 1.1 0.5 Figure 8. FACS analysis of the percent of NK cell in peripheral blood of DTREGFP/+ 0.0 MC38 tumor-bearing Ncr1 mice and C57BL/6 mice (A and B). (In Pre-dose day 3 day 7 day 14 day 21 collaboration with CrownBio) Days
www.modelorg.com 6 SMOC Mouse Models For Immunology Research ) 3 3500 Ncr1-IRES-DTR-EGFP Naïve C57BL/6 3000
2500
2000
1500 Day1,3,5: DT dose 1000 administration DTREGFP/+ Figure 9. The growth of MC38 tumors in Ncr1 mice and C57BL/6
Tumor Volume±SEM (mm 500 DTREGFP/+ mice. Ncr1 mice and C57BL/6 mice were inoculated with MC38 3 DTREGFP/+ 0 colon cancer cells. After the tumors grew to 100 mm , Ncr1 mice 0 2 4 6 8 10 12 14 16 18 20 22 24 were injected with Diphtheria toxin. (In collaboration with CrownBio) Days
More mouse models
Strain Name Cat.No. Target cells
Cd19-DTREGFP NM-KI-190042 B cells
+ Cd4-DTREGFP NM-KI-190121 Cd4 T cells
+ Cd8a-DTREGFP NM-KI-190045 Cd8 T cells
Clec4f-DTRGFP NM-KI-200070 kupffer cells
Foxp3-DTREGFP NM-KI-190046 Regulatory T lymphocytes
Itgam-DTREGFP NM-KI-200066 Macrophages
Itgax-DTREGFP NM-KI-190043 Dendritic cells
Lyz2-DTREGFP NM-KI-190041 Myeloid cell lineage (monocytes, mature macrophages and granulocytes)
Ncr1-DTREGFP NM-KI-190044 Natural killer cells
SMOC Mouse Models For Immunology Research 7 www.modelorg.com Everything you need for model organisms
●● Model generation
●● Research-Ready GEM models
●● Breeding
●● Phenotyping
●● Drug screening
Shanghai Model Organisms Center, Inc. 3rd floor, Building 2, No.178 Banxia Road, Pudong New District, Shanghai, China www.modelorg.com・[email protected]