PMID- 28645528 OWN - NLM STAT- MEDLINE DCOM- 20171018 LR - 20180321 IS - 1872-7905 (Electronic) IS - 0022-1759 (Linking) VI - 449 DP - 2017 Oct TI - Long-term sustainable dendritic cell-specific depletion murine model for periodontitis research. PG - 7-14 LID - S0022-1759(17)30146-1 [pii] LID - 10.1016/j.jim.2017.06.007 [doi] AB - 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. CI - Copyright (c) 2017 Elsevier B.V. All rights reserved. FAU - Finger Stadler, Amanda AU - Finger Stadler A AD - Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: afingerstadler@augusta.edu. FAU - Patel, Mitulkumar AU - Patel M AD - Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: mpatel3@augusta.edu. FAU - Pacholczyk, Rafal AU - Pacholczyk R AD - Department of Biochemistry and Molecular Biology, Georgia Cancer Center at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: rpacholczyk@augusta.edu. FAU - Cutler, Christopher W AU - Cutler CW AD - Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: chcutler@augusta.edu. FAU - Arce, Roger M AU - Arce RM AD - Department of Periodontics, The Dental College of Georgia at Augusta University, 1120 15th Street, Augusta, GA 30912, USA. Electronic address: rarcemunoz@augusta.edu. LA - eng PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20170621 PL - Netherlands TA - J Immunol Methods JT - Journal of immunological methods JID - 1305440 RN - 0 (Diphtheria Toxin) SB - IM MH - Animals MH - Bone Marrow Cells/immunology MH - Dendritic Cells/*immunology/pathology MH - Diphtheria Toxin/administration & dosage/immunology MH - *Disease Models, Animal MH - Mice MH - Mice, Inbred C57BL MH - Mice, Knockout MH - Mice, Transgenic MH - Periodontitis/*immunology MH - Spleen/cytology/immunology MH - Time Factors OTO - NOTNLM OT - Animal models OT - Dendritic cells OT - Diphtheria toxin OT - Knockout OT - Mice EDAT- 2017/06/25 06:00 MHDA- 2017/10/19 06:00 CRDT- 2017/06/25 06:00 PHST- 2017/03/28 00:00 [received] PHST- 2017/05/11 00:00 [revised] PHST- 2017/06/19 00:00 [accepted] PHST- 2017/06/25 06:00 [pubmed] PHST- 2017/10/19 06:00 [medline] PHST- 2017/06/25 06:00 [entrez] AID - S0022-1759(17)30146-1 [pii] AID - 10.1016/j.jim.2017.06.007 [doi] PST - ppublish SO - J Immunol Methods. 2017 Oct;449:7-14. doi: 10.1016/j.jim.2017.06.007. Epub 2017 Jun 21.