Bioluminescence for in vivo detection of cell-type-specific inflammation in a mouse model of uveitis.
Animals
Animals, Genetically Modified
B-Lymphocytes
/ chemistry
Disease Models, Animal
Feasibility Studies
Female
Genes, Reporter
/ genetics
Humans
Luciferases
/ chemistry
Luminescent Measurements
/ methods
Male
Mice
Myeloid Cells
/ chemistry
T-Lymphocytes
/ chemistry
Tomography, Optical Coherence
Uvea
/ cytology
Uveitis
/ diagnosis
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
09 07 2020
09 07 2020
Historique:
received:
04
02
2020
accepted:
01
06
2020
entrez:
11
7
2020
pubmed:
11
7
2020
medline:
22
12
2020
Statut:
epublish
Résumé
This study reports the use of cell-type-specific in vivo bioluminescence to measure intraocular immune cell population dynamics during the course of inflammation in a mouse model of uveitis. Transgenic lines expressing luciferase in inflammatory cell subsets (myeloid cells, T cells, and B cells) were generated and ocular bioluminescence was measured serially for 35 days following uveitis induction. Ocular leukocyte populations were identified using flow cytometry and compared to the ocular bioluminescence profile. Acute inflammation is neutrophilic (75% of ocular CD45 + cells) which is reflected by a significant increase in ocular bioluminescence in one myeloid reporter line on day 2. By day 7, the ocular T cell population increases to 50% of CD45 + cells, leading to a significant increase in ocular bioluminescence in the T cell reporter line. While initially negligible (< 1% of CD45 + cells), the ocular B cell population increases to > 4% by day 35. This change is reflected by a significant increase in the ocular bioluminescence of the B cell reporter line starting on day 28. Our data demonstrates that cell-type-specific in vivo bioluminescence accurately detects changes in multiple intraocular immune cell populations over time in experimental uveitis. This assay could also be useful in other inflammatory disease models.
Identifiants
pubmed: 32647297
doi: 10.1038/s41598-020-68227-4
pii: 10.1038/s41598-020-68227-4
pmc: PMC7347586
doi:
Substances chimiques
Luciferases
EC 1.13.12.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
11377Subventions
Organisme : NEI NIH HHS
ID : P30-001730
Pays : United States
Organisme : NEI NIH HHS
ID : R01 EY030431
Pays : United States
Organisme : NEI NIH HHS
ID : P30 EY001730
Pays : United States
Organisme : NEI NIH HHS
ID : K08 EY023998
Pays : United States
Références
Chu, C. J. et al. Multimodal analysis of ocular inflammation using the endotoxin-induced uveitis mouse model. Dis. Model. Mech. 9, 473–481 (2016).
doi: 10.1242/dmm.022475
Pepple, K. L., Wilson, L. & Van Gelder, R. N. Comparison of aqueous and vitreous lymphocyte populations from two rat models of experimental uveitis. Invest. Ophthalmol. Vis. Sci. 59, 2504–2511 (2018).
doi: 10.1167/iovs.18-24192
Liyanage, S. E. et al. Flow cytometric analysis of inflammatory and resident myeloid populations in mouse ocular inflammatory models. Exp. Eye Res. 151, 160–170 (2016).
doi: 10.1016/j.exer.2016.08.007
Gutowski, M. B., Wilson, L., Van Gelder, R. N. & Pepple, K. L. In vivo bioluminescence imaging for longitudinal monitoring of inflammation in animal models of uveitis. Invest. Ophthalmol. Vis. Sci. 58, 1521–1528 (2017).
doi: 10.1167/iovs.16-20824
Pepple, K. L., Choi, W. J., Wilson, L., Van Gelder, R. N. & Wang, R. K. Quantitative assessment of anterior segment inflammation in a rat model of uveitis using spectral-domain optical coherence tomography. Invest. Ophthalmol. Vis. Sci. 57, 3567–3575 (2016).
doi: 10.1167/iovs.16-19276
Gross, S. et al. Bioluminescence imaging of myeloperoxidase activity in vivo. Nat. Med. 15, 455–461 (2009).
doi: 10.1038/nm.1886
Luker, K. E. & Luker, G. D. Bioluminescence imaging of reporter mice for studies of infection and inflammation. Antiviral Res. 86, 93–100 (2010).
doi: 10.1016/j.antiviral.2010.02.002
Koo, V., Hamilton, P. W. & Williamson, K. Non-invasive in vivo imaging in small animal research. Cell. Oncol. 28, 127–139 (2006).
pubmed: 16988468
pmcid: 4617494
Zinn, K. R. et al. Noninvasive bioluminescence imaging in small animals. ILAR J. 49, 103–115 (2008).
doi: 10.1093/ilar.49.1.103
Safran, M. et al. Mouse reporter strain for noninvasive bioluminescent imaging of cells that have undergone Cre-mediated recombination. Mol. Imaging 2, 297–302 (2003).
doi: 10.1162/153535003322750637
Mezzanotte, L., Root, M., Karatas, H., Goun, E. A. & Löwik, C. W. In Vivo molecular bioluminescence imaging: new tools and applications. Trends Biotechnol. 35, 640–652 (2017).
doi: 10.1016/j.tibtech.2017.03.012
Luker, G. D. et al. Noninvasive bioluminescence imaging of herpes simplex virus type 1 infection and therapy in living mice. J. Virol. 76, 12149–12161 (2002).
doi: 10.1128/JVI.76.23.12149-12161.2002
Ezra-Elia, R. et al. Can an in vivo imaging system be used to determine localization and biodistribution of AAV5-mediated gene expression following subretinal and intravitreal delivery in mice?. Exp. Eye Res. 176, 227–234 (2018).
doi: 10.1016/j.exer.2018.08.021
Ji, X. et al. Noninvasive visualization of retinoblastoma growth and metastasis via bioluminescence imaging. Invest. Ophthalmol. Vis. Sci. 50, 5544–5551 (2009).
doi: 10.1167/iovs.08-3258
Sauer, B. & Henderson, N. Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proc. Natl. Acad. Sci. USA. 85, 5166–5170 (1988).
doi: 10.1073/pnas.85.14.5166
Rickert, R. C., Roes, J. & Rajewsky, K. B lymphocyte-specific, Cre-mediated mutagenesis in mice. Nucleic Acids Res. 25, 1317–1318 (1997).
doi: 10.1093/nar/25.6.1317
Hennet, T., Hagen, F. K., Tabak, L. A. & Marth, J. D. T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site-directed recombination. Proc. Natl. Acad. Sci. USA. 92, 12070–12074 (1995).
doi: 10.1073/pnas.92.26.12070
Clausen, B. E., Burkhardt, C., Reith, W., Renkawitz, R. & Förster, I. Conditional gene targeting in macrophages and granulocytes using LysMcre mice. Transgenic Res. 8, 265–277 (1999).
doi: 10.1023/A:1008942828960
Wang, K., Wei, G. & Liu, D. CD19: A biomarker for B cell development, lymphoma diagnosis and therapy. Exp. Hematol. Oncol. 1, 36 (2012).
doi: 10.1186/2162-3619-1-36
Bielory, L. Allergic and immunologic disorders of the eye Part I: immunology of the eye. J. Allergy Clin. Immunol. 106, 805–816 (2000).
doi: 10.1067/mai.2000.111029
Pepple, K. L. et al. Primed mycobacterial uveitis (PMU): Histologic and cytokine characterization of a model of uveitis in rats. Invest. Ophthalmol. Vis. Sci. 56, 8438–8448 (2015).
doi: 10.1167/iovs.15-17523
Mruthyunjaya, P. et al. Efficacy of low-release-rate fluocinolone acetonide intravitreal implants to treat experimental uveitis. Arch. Ophthalmol. 124, 1012–1018 (2006).
doi: 10.1001/archopht.124.7.1012
Cousins, S. W. T Cell Activation within Different Intraocular Compartments during Experimental Uveitis1. in 150–155 (1992).
Cheng, C. K., Berger, A. S., Pearson, P. A., Ashton, P. & Jaffe, G. J. Intravitreal sustained-release dexamethasone device in the treatment of experimental uveitis. Invest. Ophthalmol. Vis. Sci. 36, 442–453 (1995).
pubmed: 7843913
Lowder, C. et al. Dexamethasone intravitreal implant for noninfectious intermediate or posterior uveitis. Arch. Ophthalmol. 129, 545–553 (2011).
doi: 10.1001/archophthalmol.2010.339
Jaffe, G. J. et al. Fluocinolone acetonide implant (Retisert) for noninfectious posterior uveitis: thirty-four–week results of a multicenter randomized clinical study. Ophthalmology 113, 1020–1027 (2006).
doi: 10.1016/j.ophtha.2006.02.021
Callanan, D. G., Jaffe, G. J., Martin, D. F., Pearson, P. A. & Comstock, T. L. Treatment of posterior uveitis with a fluocinolone acetonide implant. Arch. Ophthalmol. 126, 1191–1201 (2008).
doi: 10.1001/archopht.126.9.1191
Jaffe, G. J. et al. Intravitreal sustained-release cyclosporine in the treatment of experimental uveitis. Ophthalmology 105, 46–56 (1998).
doi: 10.1016/S0161-6420(98)91176-9
Czupryna, J. & Tsourkas, A. Firefly luciferase and RLuc8 exhibit differential sensitivity to oxidative stress in apoptotic cells. PLoS ONE 6, e20073 (2011).
doi: 10.1371/journal.pone.0020073
Njus, D., Baldwin, T. O. & Hastings, J. W. A sensitive assay for proteolytic enzymes using bacterial luciferase as a substrate. Anal. Biochem. 61, 280–287 (1974).
doi: 10.1016/0003-2697(74)90356-X
Pham, C. T. N. Neutrophil serine proteases: specific regulators of inflammation. Nat. Rev. Immunol. 6, 541–550 (2006).
doi: 10.1038/nri1841
Grote, J. et al. Identification of poly(ADP-ribose)polymerase-1 and Ku70/Ku80 as transcriptional regulators of S100A9 gene expression. BMC Mol. Biol. 7, 48 (2006).
doi: 10.1186/1471-2199-7-48
Caspi, R. R. et al. T cell lines mediating experimental autoimmune uveoretinitis (EAU) in the rat. J. Immunol. 136, 928–933 (1986).
pubmed: 2416842
Shao, H. et al. Severe chronic experimental autoimmune uveitis (EAU) of the C57BL/6 mouse induced by adoptive transfer of IRBP1–20-specific T cells. Exp. Eye Res. 82, 323–331 (2006).
doi: 10.1016/j.exer.2005.07.008
Pracht, K. et al. A new staining protocol for detection of murine antibody-secreting plasma cell subsets by flow cytometry. Eur. J. Immunol. 47, 1389–1392 (2017).
doi: 10.1002/eji.201747019
Tellier, J. & Nutt, S. L. Standing out from the crowd: How to identify plasma cells. Eur. J. Immunol. 47, 1276–1279 (2017).
doi: 10.1002/eji.201747168
Smith, J. R., Stempel, A. J., Bharadwaj, A. & Appukuttan, B. Involvement of B cells in non-infectious uveitis. Clin. Transl. Immunol. 5, e63 (2016).
doi: 10.1038/cti.2016.2
Kielczewski, J. L., Horai, R., Jittayasothorn, Y., Chan, C.-C. & Caspi, R. R. Tertiary lymphoid tissue forms in retinas of mice with spontaneous autoimmune uveitis and has consequences on visual function. J. Immunol. 196, 1013–1025 (2016).
doi: 10.4049/jimmunol.1501570
Kleinwort, K. J. H. et al. Immunological characterization of intraocular lymphoid follicles in a spontaneous recurrent uveitis model. Invest. Ophthalmol. Vis. Sci. 57, 4504–4511 (2016).
doi: 10.1167/iovs.16-19787
Epps, S. J. et al. Features of ectopic lymphoid-like structures in human uveitis. Exp. Eye Res. 191, 107901 (2019).
doi: 10.1016/j.exer.2019.107901
Heng, J. S. et al. Comprehensive analysis of a mouse model of spontaneous uveoretinitis using single-cell RNA sequencing. Proc. Natl. Acad. Sci. USA https://doi.org/10.1073/pnas.1915571116 (2019).
doi: 10.1073/pnas.1915571116
pubmed: 31843893