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

Amanda Finger Stadler, Mitulkumar Patel, Rafal W Pacholczyk, Christopher W Cutler, Roger Mauricio Arce Munoz

Research output: Contribution to journalArticle

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 20 ng/bw, followed and maintained with doses of 10 ng/bm every 3 days for up to 4 weeks 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, 24 h, 1 and 4 weeks. 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 24 h and depletion maintained for up to 4 weeks, as compared to blank (PBS) controls. Circulating cDCs in spleen demonstrated a non-significant trend to deplete in 1 week with high variability among mice. GT also showed a similar non-significant trend to deplete in 24 h. 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.

Original languageEnglish (US)
Pages (from-to)7-14
Number of pages8
JournalJournal of Immunological Methods
Volume449
DOIs
StatePublished - Oct 1 2017

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Periodontitis
Dendritic Cells
Research
Alveolar Bone Loss
Diphtheria Toxin
Spleen
Bone Marrow
Antigen-Presenting Cells
Neutrophils
Homeostasis
Animal Models

Keywords

  • Animal models
  • Dendritic cells
  • Diphtheria toxin
  • Knockout
  • Mice

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Long-term sustainable dendritic cell-specific depletion murine model for periodontitis research. / Finger Stadler, Amanda; Patel, Mitulkumar; Pacholczyk, Rafal W; Cutler, Christopher W; Arce Munoz, Roger Mauricio.

In: Journal of Immunological Methods, Vol. 449, 01.10.2017, p. 7-14.

Research output: Contribution to journalArticle

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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 20 ng/bw, followed and maintained with doses of 10 ng/bm every 3 days for up to 4 weeks 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, 24 h, 1 and 4 weeks. 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 24 h and depletion maintained for up to 4 weeks, as compared to blank (PBS) controls. Circulating cDCs in spleen demonstrated a non-significant trend to deplete in 1 week with high variability among mice. GT also showed a similar non-significant trend to deplete in 24 h. 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.",
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