Foxp3 gfp mice

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Foxp3-GFP mice are a transgenic mouse model that expresses the green fluorescent protein (GFP) under the control of the Foxp3 gene promoter. Foxp3 is a transcription factor that is specifically expressed in regulatory T cells (Tregs). The expression of GFP in these mice allows for the identification and isolation of Tregs.

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17 protocols using foxp3 gfp mice

Murine Immunology Research Protocol

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Six to eight week-old C57BL6/J males, C57BL6/J-Tg (TcraTcrb)1100Mjb/J (OT-I) and Foxp3^GFP mice were purchased from Jackson Laboratories (Bar Harbor, ME) and housed in the specific pathogen-free Emory University Vivarium in compliance with the Institutional Animal Care and Use Committee (IACUC) and NIH guidelines for the care and use of laboratory animals.

Tedesco D., Thapa M., Gumber S., Elrod E.J., Rahman K., Ibegbu C.C., Magliocca J.F., Adams A.B., Anania F, & Grakoui A. (2016). CD4+ Foxp3+ T cells promote aberrant IgG production and maintain CD8+ T cell suppression during chronic liver disease. Hepatology (Baltimore, Md.), 65(2), 661-677.

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Microbiome Manipulation in Mice

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Animal procedures were approved by the Harvard Medical Area (HMA) Standing Committee on Animals. C57BL/6J mice and Foxp3^GFP+ mice (Stock No. 023800) were from The Jackson Laboratory and acclimated in the local animal facility for at least two weeks prior to study initiation. Otherwise specified, all mice used were 6–8 weeks old at the initiation of study. For all experiments (fecal transplantation, fecal RNA transplantation, synthetic microRNA administration, bacteria administration), mice of same age and gender were ear-tagged and randomly allocated into groups and co-housed. Mice were housed under specific pathogen-free conditions at the Harvard Institutes of Medicine and the Hale Building for Transformative Medicine at Brigham and Women’s Hospital. Mice that received oral administration of Akkermansia muciniphila (A. muciniphila) or E. coli were housed in BSL-2N facility. In some experiments, germ-free C57BL/6 mice (Taconic Biosciences, Inc) were kept in the germ-free container at the Hale Building for Transformative Medicine at Brigham and Women’s Hospital and were used immediately upon arrival. Fecal specimens were collected immediately upon defecation, snap frozen, and stored at −80 °C for analysis.

Liu S., Rezende R.M., Moreira T.G., Tankou S.K., Cox L.M., Wu M., Song A., Dhang F.H., Wei Z., Costamagna G, & Weiner H.L. (2019). Oral administration of miR-30d from feces of MS patients suppresses MS-like symptoms in mice by expanding Akkermansia muciniphila. Cell host & microbe, 26(6), 779-794.e8.

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Atherosclerosis Progression in Transgenic Mice

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All mice used were on a C57BL/6 background. Except in Figure 6, male mice were used in all experiments. ApoE^–/– mice (stock no. 002052), Il12a^–/– mice (stock no. 002692), and Foxp3-GFP mice (stock no. 006772) were purchased from the Jackson Laboratory. Il12a^–/–/ ApoE^–/– (IL-35P35^–/–/ApoE^–/–) and Foxp3-GFP/ApoE^–/– were crossbred in our laboratory. All mice were maintained on a chow diet until 8 weeks old. For HFD feeding, they were given HFD (TD.88137, Harlan) composed of cholesterol (0.2%, w/w) and fat (21.2%, w/w), started from 8 weeks.

Shao Y., Yang W.Y., Saaoud F., Drummer C I.V., Sun Y., Xu K., Lu Y., Shan H., Shevach E.M., Jiang X., Wang H, & Yang X. (2021). IL-35 promotes CD4+Foxp3+ Tregs and inhibits atherosclerosis via maintaining CCR5-amplified Treg-suppressive mechanisms. JCI Insight, 6(19), e152511.

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Generation and Maintenance of Mouse Models

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C57BL/6 and FoxP3^GFP mice were obtained from Jackson laboratory. Irf8^−/− and Ifnar1^−/− mice were kindly provided by Dr. Harinder Singh and Dr. Edith Jannsen at Cincinnati Children’s Hospital Medical Center (CCHMC), respectively. Animals were maintained by the Department of Laboratory Animal Medicine, University of Cincinnati, accredited by the American Association for Accreditation of Laboratory Animal Care (Frederick, MD). All animal experiments were in accordance with Animal Welfare Act guidelines of National Institutes of Health.

George M.M., Vignesh K.S., Landero Figueroa J.A., Caruso J.A, & Deepe GS J.r. (2016). Zinc Induces Dendritic Cell Tolerogenic Phenotype and Skews Regulatory T cell – Th17 Balance. Journal of immunology (Baltimore, Md. : 1950), 197(5), 1864-1876.

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Murine models for immunology research

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Mice were kept in accordance with guidelines of the Johns Hopkins University Institutional Animal Care and Use Committee. 5c.c7 Rag^–/—and OT-II mice were purchased from Taconic. C57BL/6 mice, CD90.1 mice, CD4 Cre recombinase mice, Foxp3^GFP+ mice, and mice with loxP-flanked Raptor alleles were obtained from Jackson Laboratories. Mice with loxP-flanked Tsc2 alleles were generated by the laboratory of M. Gambello [24 (link)]. 5c.c7 Rag^–/—mice were derived from a B10.A background, while all other WT and OT II+ mice described in this manuscript originate from the C57BL/6 strain.

Pollizzi K.N., Waickman A.T., Patel C.H., Sun I.H, & Powell J.D. (2015). Cellular Size as a Means of Tracking mTOR Activity and Cell Fate of CD4+ T Cells upon Antigen Recognition. PLoS ONE, 10(4), e0121710.

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Foxp3-GFP Mice Protocol

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Foxp3-GFP mice were obtained from Jackson Laboratories and were bred according to the Experimental Animal Centre of the Third Military Medical University and housed in a specific pathogen-free facility. Mice ages 6–8 weeks were used for the experiment. The experimental protocol was approved by the Ethics Committee of the Third Military Medical University.

Cai Z., Liu H, & Wu X. (2017). Forkhead-box transcription factor 1 affects the apoptosis of natural regulatory T cells by controlling Aven expression. BMC Immunology, 18, 16.

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Genetically Modified Mice for Immunology Research

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C57BL/6J mice, SJL mice, BALB/cJ mice, B10.A mice, C57BL/6-Tg(UBC-GFP)30Scha/J mice, B6.Cg-Tg(CAG-DsRed*MST)1Nagy/J mice, CD73 KO mice, FoxP3-YFP cre mice, FoxP3-GFP mice, CXCR4-flox mice, Lep-cre mice, C;129S-Adora2atm1Jfc/J, and CXCL12-flox mice (Jackson Laboratory, Bar Harbor, ME) were housed in a specific pathogen-free environment. CXCL12-DsRed mice were kindly provided by Dr. Lei Ding (Ding and Morrison, 2013 (link)). CD39-flox mice were kindly provided by Dr. Simon C. Robson (Harvard Medical School). 7-week-old Foxp3^cre CXCR4^fl/fl mice and Foxp3^cre CXCR4^fl/wt mice were analyzed. For in vivo imaging, the mice were anaesthetized with 2–3% isoflurane and buprenorphine. The mice were sacrificed by CO₂ inhalation and cervical dislocation. Studies were conducted with approval from Institutional Review Boards and Animal Care and Use Committees at Columbia University.

Hirata Y., Furuhashi K., Ishii H., Li H.W., Pinho S., Ding L., Robson S.C., Frenette P.S, & Fujisaki J. (2018). CD150high bone marrow Tregs maintain hematopoietic stem cell quiescence and immune privilege via adenosine. Cell stem cell, 22(3), 445-453.e5.

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Mice Sourcing and Housing for Experimental Studies

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C57BL/6 mice (10–12 weeks old, equal number of males and females) were purchased from the Hubei Provincial Center for Disease Control and Prevention (Wuhan, China). Twelve-week-old male Foxp3^GFP mice were obtained from Jackson Laboratory (Bar Harbor, ME, USA). The mice were housed at the Center for Experimental Animals, Tongji Hospital, Huazhong University of Science and Technology, for at least 1 week before any experiment. All experiments were approved by the Institutional Animal Care and Use Committee of Tongji Hospital, Huazhong University of Science and Technology (2015 IACUC: number 595).

Fu X., Sun F., Wang F., Zhang J., Zheng B., Zhong J., Yue T., Zheng X., Xu J.F, & Wang C.Y. (2017). Aloperine Protects Mice against DSS-Induced Colitis by PP2A-Mediated PI3K/Akt/mTOR Signaling Suppression. Mediators of Inflammation, 2017, 5706152.

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Murine Model for Antigen-Specific Immune Response

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All mice were bred and maintained under pathogen-free conditions within American Association of Laboratory Animal Care approved facilities at the University of California, Berkeley. The internal review board and the Animal Use and Care Committee at UC Berkeley approved all procedures. C57BL/6, B6.SJL-Ptprca Pepcb/BoyJ (CD45.1), 2D2 TCR transgenic, AND TCR transgenic, B6.129P2-Il2tm1Hor/J(IL2−/−), C57BL/6-Tg(Ins2-TFRC/OVA)296Wehi/WehiJ (RIPmOVA), and Foxp3-GFP mice were purchased from Jackson. 2D2×Rag2^−/− mice were generated by crossing 2D2 mice with Rag2^−/− mice purchased from Taconic. OT-II×Rag2^−/− TCR transgenic mice were purchased from Taconic. F5×Rag2^−/− mice have been described previously^{45 (link)}. Endotoxin-free soluble ovalbumin was purchased from Invivogen, and injections were performed via i.v. injection, followed by sacrifice 1 h later.

Weist B.M., Kurd N., Boussier J., Chan S.W, & Robey E.A. (2015). Thymic regulatory T cell niche size is dictated by limiting interleukin 2 from antigen-bearing dendritic cells and feedback competition. Nature immunology, 16(6), 635-641.

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Transgenic Mouse Models for Immunology

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All mice were bred and housed under specific pathogen free (SPF) conditions. All animal experiments were approved by TSRI Animal Care and Use Committee. CD45.2 (WT), CD45.1^+/+, Thy1.1^+/+ and Foxp3-GFP mice in C57BL/6J genetic background, BALB/c mice, OT-I mice and P14 mice were purchased from the Jackson Laboratory. OT-II mice were from Dr. James C. Paulson, Scripps Research. OT-I^+/−CD45.1^+/− and OT-I^+/−Thy1.1^+/− mice were generated by cross breeding OT-I with CD45.1^+/+ and Thy1.1^+/+ C57BL/6J mice, respectively. Both male and female mice of 6–12 weeks of age were used for all experiments.

Liu Z., Li J.P., Chen M., Wu M., Shi Y., Li W., Teijaro J.R, & Wu P. (2020). Detecting Tumor Antigen-specific T cells via Interaction Dependent Fucosyl-biotinylation. Cell, 183(4), 1117-1133.e19.

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