Establishment of a Human Microbiome- and Immune System-reconstituted Dual-humanized Mouse Model

Overview

Journal Exp Anim

Date 2023 Apr 5

PMID 37019665

Authors

Yuyo Ka

Ryoji Ito

Ryoko Nozu

Kayo Tomiyama

Masami Ueno

Tomoyuki Ogura

Riichi Takahashi

Affiliations

Soon will be listed here.

Abstract

Humanized mice are widely used to study the human immune system in vivo and investigate therapeutic targets for various human diseases. Immunodeficient NOD/Shi-scid-IL2rγ (NOG) mice transferred with human hematopoietic stem cells are a useful model for studying human immune systems and analyzing engrafted human immune cells. The gut microbiota plays a significant role in the development and function of immune cells and the maintenance of immune homeostasis; however, there is currently no available animal model that has been reconstituted with human gut microbiota and immune systems in vivo. In this study, we established a new model of CD34 cell-transferred humanized germ-free NOG mice using an aseptic method. Flow cytometric analysis revealed that the germ-free humanized mice exhibited a lower level of human CD3 T cells than the SPF humanized mice. Additionally, we found that the human CD3 T cells slightly increased after transplanting human gut microbiota into the germ-free humanized mice, suggesting that the human microbiota supports T cell proliferation or maintenance in humanized mice colonized by the gut microbiota. Consequently, the dual-humanized mice may be useful for investigating the physiological role of the gut microbiota in human immunity in vivo and for application as a new humanized mouse model in cancer immunology.

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References

Katoh K, Misawa K, Kuma K, Miyata T . MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res. 2002; 30(14):3059-66. PMC: 135756. DOI: 10.1093/nar/gkf436. View

Ito R, Takahashi T, Katano I, Ito M . Current advances in humanized mouse models. Cell Mol Immunol. 2012; 9(3):208-14. PMC: 4012844. DOI: 10.1038/cmi.2012.2. View

Ka Y, Ogura T, Tomiyama K, Ueno M, Nozu R, Tsuruzono N . Creation of an experimental rearing environment for microbiome animal research using an individually ventilated cage system and bioBUBBLE enclosure. Exp Anim. 2020; 70(2):177-184. PMC: 8150247. DOI: 10.1538/expanim.20-0129. View

Ito M, Hiramatsu H, Kobayashi K, Suzue K, Kawahata M, Hioki K . NOD/SCID/gamma(c)(null) mouse: an excellent recipient mouse model for engraftment of human cells. Blood. 2002; 100(9):3175-82. DOI: 10.1182/blood-2001-12-0207. View

Ekmekciu I, von Klitzing E, Fiebiger U, Escher U, Neumann C, Bacher P . Immune Responses to Broad-Spectrum Antibiotic Treatment and Fecal Microbiota Transplantation in Mice. Front Immunol. 2017; 8:397. PMC: 5395657. DOI: 10.3389/fimmu.2017.00397. View

Kennedy E, King K, Baldridge M . Mouse Microbiota Models: Comparing Germ-Free Mice and Antibiotics Treatment as Tools for Modifying Gut Bacteria. Front Physiol. 2018; 9:1534. PMC: 6220354. DOI: 10.3389/fphys.2018.01534. View

Wege A, Melkus M, Denton P, Estes J, Garcia J . Functional and phenotypic characterization of the humanized BLT mouse model. Curr Top Microbiol Immunol. 2008; 324:149-65. DOI: 10.1007/978-3-540-75647-7_10. View

Ivanov I, Atarashi K, Manel N, Brodie E, Shima T, Karaoz U . Induction of intestinal Th17 cells by segmented filamentous bacteria. Cell. 2009; 139(3):485-98. PMC: 2796826. DOI: 10.1016/j.cell.2009.09.033. View

Bolyen E, Rideout J, Dillon M, Bokulich N, Abnet C, Al-Ghalith G . Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nat Biotechnol. 2019; 37(8):852-857. PMC: 7015180. DOI: 10.1038/s41587-019-0209-9. View

10.

Pushalkar S, Hundeyin M, Daley D, Zambirinis C, Kurz E, Mishra A . The Pancreatic Cancer Microbiome Promotes Oncogenesis by Induction of Innate and Adaptive Immune Suppression. Cancer Discov. 2018; 8(4):403-416. PMC: 6225783. DOI: 10.1158/2159-8290.CD-17-1134. View

11.

Atarashi K, Tanoue T, Ando M, Kamada N, Nagano Y, Narushima S . Th17 Cell Induction by Adhesion of Microbes to Intestinal Epithelial Cells. Cell. 2015; 163(2):367-80. PMC: 4765954. DOI: 10.1016/j.cell.2015.08.058. View

12.

Mauras A, Wopereis H, Yeop I, Esber N, Delannoy J, Labellie C . Gut microbiota from infant with cow's milk allergy promotes clinical and immune features of atopy in a murine model. Allergy. 2019; 74(9):1790-1793. PMC: 6790679. DOI: 10.1111/all.13787. View

13.

Callahan B, McMurdie P, Rosen M, Han A, Johnson A, Holmes S . DADA2: High-resolution sample inference from Illumina amplicon data. Nat Methods. 2016; 13(7):581-3. PMC: 4927377. DOI: 10.1038/nmeth.3869. View

14.

Zheng J, Wong C, Liu J, Luo X, Zhou W, Chen Y . Glucose metabolism inhibitor PFK-015 combined with immune checkpoint inhibitor is an effective treatment regimen in cancer. Oncoimmunology. 2022; 11(1):2079182. PMC: 9191911. DOI: 10.1080/2162402X.2022.2079182. View

15.

Watanabe M, Zahidunnabi Dewan M, Okamura T, Sasaki M, Itoh K, Higashihara M . A novel NF-kappaB inhibitor DHMEQ selectively targets constitutive NF-kappaB activity and induces apoptosis of multiple myeloma cells in vitro and in vivo. Int J Cancer. 2004; 114(1):32-8. DOI: 10.1002/ijc.20688. View

16.

Mitsuoka T, SEGA T, Yamamoto S . [Improved methodology of qualitative and quantitative analysis of the intestinal flora of man and animals]. Zentralbl Bakteriol Orig. 1965; 195(4):455-69. View

17.

Nozu R, Ueno M, Hayashimoto N . Composition of fecal microbiota of laboratory mice derived from Japanese commercial breeders using 16S rRNA gene clone libraries. J Vet Med Sci. 2016; 78(6):1045-50. PMC: 4937141. DOI: 10.1292/jvms.15-0454. View

18.

Liang M, Liwen Z, Jianguo S, Juan D, Fei D, Yin Z . Fecal Microbiota Transplantation Controls Progression of Experimental Autoimmune Hepatitis in Mice by Modulating the TFR/TFH Immune Imbalance and Intestinal Microbiota Composition. Front Immunol. 2021; 12:728723. PMC: 8667314. DOI: 10.3389/fimmu.2021.728723. View

19.

Allam A, Majji S, Peachman K, Jagodzinski L, Kim J, Ratto-Kim S . TFH cells accumulate in mucosal tissues of humanized-DRAG mice and are highly permissive to HIV-1. Sci Rep. 2015; 5:10443. PMC: 4451806. DOI: 10.1038/srep10443. View

20.

Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P . The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res. 2012; 41(Database issue):D590-6. PMC: 3531112. DOI: 10.1093/nar/gks1219. View