Long-term sustainable dendritic cell-specific depletion murine model for periodontitis research

Amanda Finger Stadler; Mitulkumar Patel; Rafal Pacholczyk; Christopher W Cutler; Roger M Arce

doi:10.1016/j.jim.2017.06.007

Long-term sustainable dendritic cell-specific depletion murine model for periodontitis research

J Immunol Methods. 2017 Oct:449:7-14. doi: 10.1016/j.jim.2017.06.007. Epub 2017 Jun 21.

Authors

Amanda Finger Stadler¹, Mitulkumar Patel², Rafal Pacholczyk³, Christopher W Cutler⁴, Roger M Arce⁵

Affiliations

¹ Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: afingerstadler@augusta.edu.
² Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: mpatel3@augusta.edu.
³ Department of Biochemistry and Molecular Biology, Georgia Cancer Center at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: rpacholczyk@augusta.edu.
⁴ Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: chcutler@augusta.edu.
⁵ Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: rarcemunoz@augusta.edu.

PMID: 28645528
DOI: 10.1016/j.jim.2017.06.007

Abstract

Dendritic cells (DCs) are specialized antigen-presenting cells that play a pivotal role in the pathogenesis of periodontitis. The use of animal models to study the role of DCs in periodontitis has been limited by lack of a method for sustained depletion of DCs. Hence, the objectives of this study were to validate the zDC-DTR knockin mouse model of conventional DCs (cDCs) depletion, as well as to investigate whether this depletion could be sustained long enough to induce alveolar bone loss in this model. zDC-DTR mice were treated with different dose regimens of diphtheria toxin (DT) to determine survival rate. A loading DT dose of 20ng/bw, followed and maintained with doses of 10ng/bm every 3days for up to 4weeks demonstrated 80% survival. Animals were weighed weekly and peripheral blood was obtained to confirm normal neutrophil counts. Five animals per group were euthanized at baseline, 24h, 1 and 4weeks. Bone marrow (BM), spleen (SP) and gingival tissue (GT) were harvested, and cells were isolated, separated and stained for Pre-DCs precursors (CD45R^-MHCII⁺CD11c⁺Flt3⁺CD172a⁺) in BM, cDCs (CD11c⁺MHCII⁺CD209⁺) in spleen, and DCs in GT (CD45R⁺MHCII⁺CD11c⁺ DC-SIGN/CD209⁺). Pre-DCs in BM were significantly depleted at 24h and depletion maintained for up to 4weeks, as compared to blank (PBS) controls. Circulating cDCs in spleen demonstrated a non-significant trend to deplete in 1week with high variability among mice. GT also showed a similar non-significant trend to deplete in 24h. The zDC-DTR model seems to be viable for evaluating the role of DCs immune homeostasis disruption and alveolar bone loss pathogenesis in response to long-term oral infection.

Keywords: Animal models; Dendritic cells; Diphtheria toxin; Knockout; Mice.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
Bone Marrow Cells / immunology
Dendritic Cells / immunology*
Dendritic Cells / pathology
Diphtheria Toxin / administration & dosage
Diphtheria Toxin / immunology
Disease Models, Animal*
Mice
Mice, Inbred C57BL
Mice, Knockout
Mice, Transgenic
Periodontitis / immunology*
Spleen / cytology
Spleen / immunology
Time Factors

Substances

Diphtheria Toxin