Accelerated chemokine receptor 7-mediated dendritic cell migration in Runx3 knockout mice and the spontaneous development of asthma-like disease - PubMed (original) (raw)
Comparative Study
. 2005 Jul 26;102(30):10598-603.
doi: 10.1073/pnas.0504787102. Epub 2005 Jul 18.
Affiliations
- PMID: 16027362
- PMCID: PMC1180803
- DOI: 10.1073/pnas.0504787102
Comparative Study
Accelerated chemokine receptor 7-mediated dendritic cell migration in Runx3 knockout mice and the spontaneous development of asthma-like disease
Ofer Fainaru et al. Proc Natl Acad Sci U S A. 2005.
Abstract
The Runx3 transcription factor is a key regulator of lineage-specific gene expression in several developmental pathways and could also be involved in autoimmunity. We report that, in dendritic cells (DC), Runx3 regulates TGFbeta-mediated transcriptional attenuation of the chemokine receptor CCR7. When Runx3 is lost, i.e., in Runx3 knockout mice, expression of CCR7 is enhanced, resulting in increased migration of alveolar DC to the lung-draining lymph nodes. This increased DC migration and the consequent accumulation of activated DC in draining lymph nodes is associated with the development of asthma-like features, including increased serum IgE, hypersensitivity to inhaled bacterial lipopolysaccharide, and methacholine-induced airway hyperresponsiveness. The enhanced migration of DC in the knockout mice could be blocked in vivo by anti-CCR7 antibodies and by the drug Ciglitazone, known to inhibit CCR7 expression. The data indicate that Runx3 transcriptionally regulates CCR7 and that, when absent, the dysregulated expression of CCR7 in DC plays a role in the etiology of asthmatic conditions that recapitulate clinical symptoms of the human disease. Interestingly, human RUNX3 resides in a region of chromosome 1p36 that contains susceptibility genes for asthma and hypersensitivity against environmental antigens. Thus, mutations in RUNX3 may be associated with increased sensitivity to asthma development.
Figures
Fig. 1.
Human asthma hallmarks in Runx3 KO mice. (A) Increase of AHR in the KO mice. AHR was determined in 4- to 9-week-old KO mice and WT littermates as described in Materials and Methods. Results are presented as fold increase in enhanced pause (Penh) [a parameter related to pulmonary resistance (41)] relative to baseline value. Data represent the mean ± SEM of 15 mice per group and disclose a significant difference between WT and KO mice (Student's two-tailed t test; *, P = 0.0005). (B) Increase in serum and BAL IgE in the KO mice. Total IgE in serum and BAL of 4- to 6-week-old KO mice (n = 7) and WT littermates (n = 6) was quantitated by ELISA (Mouse IgE ELISA Quantization Kit, E90-115, Bethyl Laboratories, Montgomery, TX) according to the manufacturer's recommended conditions. Values in ng/ml disclose significant differences between WT and KO mice (Student's two-tailed t test; *, P = 0.026; **, P = 0.014). (C) Increased expression of FcεR receptors on DC from KO thoracic LN. Single-cell suspension was obtained from LN of WT and KO mice (n = 3), subjected to acid elution of occupied FcεR receptor, incubated with IgE-FITC conjugate and with anti CD11c, and subjected to FACS analysis. CD11c+ cells were gated, and FcεR level was determined (D) Increased responsiveness of KO mice to inhaled LPS. Runx3 KO mice and WT littermates (n = 4) were anesthetized and subjected to intranasal inhalation of 25 μl of PBS containing 0.1 μg of LPS or 25 μl of PBS alone. Sixteen hours later mice were killed and BAL was obtained and analyzed by FACS and, after cytospin, by Giemsa staining. Circled are specific populations: CD11c+ DC (circles 1 and 2), neutrophils (circle 3), and eosinophils (circle 4), as was also confirmed by Giemsa staining (data not shown).
Fig. 2.
Dysregulated expression of CCR7 in Runx3 KO DC. (A) Impaired TGFβ-dependent inhibition of CCR7 transcription in KO BMDC. WT and Runx3 KO BMDC were grown in the presence or absence of TGFβ (10 ng/ml). At day 6, cells were induced to undergo maturation by LPS, and, at day 7, RNA was prepared and analyzed by RT-PCR. (B) Impaired TGFβ-dependent inhibition of surface expression of CCR7 in KO BMDC. WT and Runx3 KO BMDC were grown and treated as in A. At day 7, cells were analyzed by FACS by using anti-CCR7 antibodies (goat anti-mouse CI0131, Capralogics). FSChighCD11c+ DC were gated, and their CCR7 expression was determined. Reduction in the level of surface CCR7 and in the number of cells expressing it was noted only in WT and not in Runx3 KO BMDC. (C and D) Increased CCR7 expression on alveolar and LN DC of Runx3 KO mice. BAL and peripheral LN cells of KO and WT mice (n = 3) were obtained and analyzed. (C) Alveolar DC (FSChigh/CD11chigh) were gated and analyzed for CCR7 expression. Of note, expression of CCR7 on the DC subpopulation CD11c+/CD11b+ present only in KO lungs is shown along with that of KO CD11c+/CD11b- DC. (D) FSChigh/CD11chigh DC of axillary and thoracic LN were gated and analyzed for CCR7 expression.
Fig. 3.
SLC-directed chemotaxis of Runx3 KO DC is not attenuated by TGFβ. (A) BMDC from WT and Runx3 KO littermates were cultured as above with or without TGFβ and, at day 6, were treated overnight with LPS. BMDC (5 × 105) were placed in a Transwell migration chamber, and SLC-dependent chemotaxis was measured as described in Materials and Methods. The mean ± SEM of three separate experiments is presented. Migration inhibition of WT DC by TGFβ was significant (*) by using the paired Student t test (P = 0.05). (B) Dermal sheaths of WT and KO mice (n = 3) were prepared, and SLC-mediated chemotactic migration was assessed. Inhibition of WT DC migration by TGFβ (*) was significant (P = 0.029) as were the responses of WT and KO DC to SLC and TGFβ (**)(P = 0.019).
Fig. 4.
Elevated CCR7 mediated in vivo trafficking of alveolar DC to the draining LN in the KO mice. (A and B) Runx3 KO (n = 7) and WT (n = 5) mice were treated by intranasal administration of CFSE to label in vivo the respiratory DC. When indicated, WT mice were treated with LPS (n = 4), and Runx3 KO mice were treated with anti-CCR7 antibody (n = 4) or with buffer only (n = 4). KO mice (n = 4) and LPS-treated WT mice (n = 3) were also treated by inhalation of Ciglitazone. Eighteen hours later, mice were killed, and single-cell suspensions of BAL, thoracic LN, and axillary LN were prepared and analyzed by FACS. (A) FSChigh/CD11c+ DC were gated (R1 and R2). Shown is representative side scatter (SSC) versus CFSE staining of DC populations in BAL and LN after the various treatments. (B) Migration index of alveolar DC to thoracic LN represents the ratio between the percentage of CFSE+ cells within the CD11c+ population in the thoracic LN and the respective value in BAL cells. Results are presented as mean ± SEM. Analysis of variance showed that the migration index of untreated KO DC was significantly higher than that of WT (*, P = 0.016). Notably, the anti-CCR7-treated KO DC migration index was similar to basal migration of WT, and Ciglitazone significantly (*, P = 0.03) reduced the migration of KO DC.
Similar articles
- Murine neuroblastoma attenuates dendritic cell cysteine cysteine receptor 7 (CCR7) expression.
Walker SR, Aboka A, Ogagan PD, Barksdale EM Jr. Walker SR, et al. J Pediatr Surg. 2005 Jun;40(6):983-7. doi: 10.1016/j.jpedsurg.2005.03.042. J Pediatr Surg. 2005. PMID: 15991182 - CC chemokine receptor-7 on dendritic cells is induced after interaction with apoptotic tumor cells: critical role in migration from the tumor site to draining lymph nodes.
Hirao M, Onai N, Hiroishi K, Watkins SC, Matsushima K, Robbins PD, Lotze MT, Tahara H. Hirao M, et al. Cancer Res. 2000 Apr 15;60(8):2209-17. Cancer Res. 2000. PMID: 10786686 - Selected natural and synthetic retinoids impair CCR7- and CXCR4-dependent cell migration in vitro and in vivo.
Villablanca EJ, Zhou D, Valentinis B, Negro A, Raccosta L, Mauri L, Prinetti A, Sonnino S, Bordignon C, Traversari C, Russo V. Villablanca EJ, et al. J Leukoc Biol. 2008 Sep;84(3):871-9. doi: 10.1189/jlb.0108047. Epub 2008 May 30. J Leukoc Biol. 2008. PMID: 18515328 - The correlation between Runx3 and bronchial asthma.
Yu Y, Wang L, Gu G. Yu Y, et al. Clin Chim Acta. 2018 Dec;487:75-79. doi: 10.1016/j.cca.2018.09.023. Epub 2018 Sep 12. Clin Chim Acta. 2018. PMID: 30218658 Review. - RUNX3-Mediated Immune Cell Development and Maturation.
Boto P, Csuth TI, Szatmari I. Boto P, et al. Crit Rev Immunol. 2018;38(1):63-78. doi: 10.1615/CritRevImmunol.2018025488. Crit Rev Immunol. 2018. PMID: 29717663 Review.
Cited by
- Lens Epithelial Explants Treated with Vitreous Humor Undergo Alterations in Chromatin Landscape with Concurrent Activation of Genes Associated with Fiber Cell Differentiation and Innate Immune Response.
Upreti A, Padula SL, Tangeman JA, Wagner BD, O'Connell MJ, Jaquish TJ, Palko RK, Mantz CJ, Anand D, Lovicu FJ, Lachke SA, Robinson ML. Upreti A, et al. Cells. 2023 Feb 3;12(3):501. doi: 10.3390/cells12030501. Cells. 2023. PMID: 36766843 Free PMC article. - Single-cell analysis of myeloid cells in HPV+ tonsillar cancer.
Jimenez DG, Altunbulakli C, Swoboda S, Sobti A, Askmyr D, Ali A, Greiff L, Lindstedt M. Jimenez DG, et al. Front Immunol. 2023 Jan 19;13:1087843. doi: 10.3389/fimmu.2022.1087843. eCollection 2022. Front Immunol. 2023. PMID: 36741389 Free PMC article. - Inducible general knockout of Runx3 profoundly reduces pulmonary cytotoxic CD8+ T cells with minimal effect on outcomes in mice following influenza infection.
Hao Q, Kundu S, Shetty S, Tucker TA, Idell S, Tang H. Hao Q, et al. Front Immunol. 2022 Oct 7;13:1011922. doi: 10.3389/fimmu.2022.1011922. eCollection 2022. Front Immunol. 2022. PMID: 36275778 Free PMC article. - Transcriptome and DNA methylome analysis of peripheral blood samples reveals incomplete restoration and transposable element activation after 3-months recovery of COVID-19.
Yin Y, Liu XZ, Tian Q, Fan YX, Ye Z, Meng TQ, Wei GH, Xiong CL, Li HG, He X, Zhou LQ. Yin Y, et al. Front Cell Dev Biol. 2022 Oct 3;10:1001558. doi: 10.3389/fcell.2022.1001558. eCollection 2022. Front Cell Dev Biol. 2022. PMID: 36263014 Free PMC article. - MicroRNA-19a Inhibition Directly and Indirectly Ameliorates Th2 Airway Inflammation in Asthma by Targeting RUNX3.
He S, Zhou J, Ma Y, Wang W, Yang J. He S, et al. Inflammation. 2023 Feb;46(1):370-387. doi: 10.1007/s10753-022-01739-5. Epub 2022 Sep 16. Inflammation. 2023. PMID: 36112239
References
- Banchereau, J. & Steinman, R. M. (1998) Nature 392, 245-252. - PubMed
- Umetsu, D. T., McIntire, J. J., Akbari, O., Macaubas, C. & DeKruyff, R. H. (2002) Nat. Immunol. 3, 715-720. - PubMed
- Wills-Karp, M. (1999) Annu. Rev. Immunol. 17, 255-281. - PubMed
- Wills-Karp, M. (2000) Immunopharmacology 48, 263-268. - PubMed
- Upham, J. W. & Stumbles, P. A. (2003) Pharmacol. Ther. 100, 75-87. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
Molecular Biology Databases