Cytokine Regulation of Bone Loss in CD18 Mice

To determine cytokine involvement in naturally occurring bone loss in CD18^−/− mice, anti-cytokine blocking antibodies, or their controls, were microinjected locally into the palatal gingiva (5 μg per site), three times weekly from the age of 4 to 18 weeks, using a 28.5-gauge MicroFine needle (BD Biosciences). Microinjections were performed on the mesial of the first molar and in the papillae between first and second and third molars on both sides of the maxilla [43 (link)]. The following anti-mouse antibodies were used: anti-IL-17A (clone 17F3) and IgG1 isotype control, both from BioXcell; anti-IL-12p35 (clone C18.2; eBioscience) and IgG2a isotype control (R&D Systems); anti-IL-12/IL-23p40 (clone C17.8) and IgG2a isotype control, both from R&D Systems; polyclonal anti-IL-23p19 and non-immune IgG control, both from R&D Systems; anti-mouse IL-6 (clone MP5-20F3), anti-TNF (clone MP6-XT22) and IgG1 isotype control, all three reagents from R&D Systems.

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Kajikawa T., Wang B., Li X., Wang H., Chavakis T., Moutsopoulos N.M, & Hajishengallis G. (2020). Activation of metabolic nuclear receptors restores periodontal tissue homeostasis in mice with leukocyte adhesion deficiency-1. Journal of leukocyte biology, 108(5), 1501-1514.

Publication 2020

MicroFine needle anti-IL-17A (clone 17F3) IgG2a

Anti Anti antibodies Bone loss Clone Cytokine Gingiva Igg1 Igg2a Il 12 Il 12p35 Il 17a Il 23p19 Isotype Maxilla Microinjections Molar Mouse Mouse il 6 Needle Third molars

Corresponding Organization : University of Pennsylvania

Other organizations : TU Dresden, National Institute of Dental and Craniofacial Research

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Variable analysis

independent variables

Anti-cytokine blocking antibodies (anti-IL-17A, anti-IL-12p35, anti-IL-12/IL-23p40, anti-IL-23p19, anti-IL-6, anti-TNF)
Isotype controls (IgG1, IgG2a, non-immune IgG)

dependent variables

Naturally occurring bone loss in CD18-/- mice

control variables

Microinjection site (mesial of the first molar and in the papillae between first and second and third molars on both sides of the maxilla)
Microinjection frequency (three times weekly from the age of 4 to 18 weeks)
Microinjection volume (5 μg per site)
Microinjection needle (28.5-gauge MicroFine needle, BD Biosciences)

controls

Positive controls: IgG1 isotype control, IgG2a isotype control, non-immune IgG control
Negative controls: Anti-cytokine blocking antibodies (anti-IL-17A, anti-IL-12p35, anti-IL-12/IL-23p40, anti-IL-23p19, anti-IL-6, anti-TNF)

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