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. 2020 Nov 26:24:100866.
doi: 10.1016/j.bbrep.2020.100866. eCollection 2020 Dec.

IFN-β signaling dampens microglia reactivity but does not prevent from light-induced retinal degeneration

Verena Behnke  1 , Thomas Langmann  1   2
Affiliations

IFN-β signaling dampens microglia reactivity but does not prevent from light-induced retinal degeneration

Verena Behnke et al. Biochem Biophys Rep. .

Abstract

Chronic activation of microglia is associated with retinal degeneration, which makes them a potential therapeutic target for retinal degenerative diseases including age-related macular degeneration (AMD). Interferon-beta (IFN-β) is a potent immune regulator, commonly used for the treatment of multiple sclerosis patients. We have previously shown that IFN-β prevents microgliosis and choroidal neovascularization in a laser model of wet AMD. Here, we hypothesized that microglia modulation via IFN-β may also dampen mononuclear phagocyte reactivity and thereby protect from retinal degeneration in a light-damage paradigm mimicking some features of dry AMD. BALB/cJ mice received intraperitoneal injections of 10,000 U IFN-β or vehicle every other day; starting at the day of exposure to 15,000 lux white light for 1 h. Systemic treatment with IFN-β partially enhanced IFN-α/β receptor (IFNAR) signaling in the retina and reduced the number of reactivated microglia in the subretinal space. However, four days after light damage neither decreased expression of complement factors nor rescue of retinal thickness was found. We conclude that IFNAR signaling modulate retinal microglia but cannot prevent strong retinal degeneration as elicited by acute white light damage.

Keywords: Age-related macular degeneration; Complement factors; Interferon-β; Light damage; Microglia.

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Conflict of interest statement

All authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1
Biological activity of human IFN-β in BV-2 mouse microglia cells. Cells were treated with LPS (50 ng/mL) and IFN-β (1000 U/mL) for 6 h qRT-PCR was performed and ΔΔCT analysis was used for quantification. ATP5B was used as reference gene. Graphs were plotted with GraphPad Prism 7.04. Bars represent mean ± SEM. Data were analyzed using Mann-Whitney U test. # depicts significance versus control. n = 6–8 (control), n = 6–12 (treated samples).
Fig. 2
Fig. 2
Expression analysis of retinal tissue. Mice were treated with IFN-β (10,000 U) or PBS as vehicle every other day starting 1 h prior to light damage until 4 d afterwards. qRT-PCR was performed and ΔΔCT analysis was used for quantification. ATP5B was used as reference gene. Graphs were plotted with GraphPad Prism 7.04. Bars represent Mean ± SEM. Data were analyzed using Kruskal-Wallis test followed by Dunn's multiple comparison. n = 23 (no light control), n = 20–22 (light + vehicle), n = 24–25 (light + IFN-â).
Fig. 3
Fig. 3
Iba-1 staining of microglia. Mice were treated with IFN-β (10,000 U) or PBS as vehicle every other day starting 1 h prior to light damage until 4 d afterwards. A Microglia were stained on retinal flat mounts using ionized calcium-binding adapter molecule 1 (Iba-1). B 4 images of the central retina were taken around the optic nerve of each eye and averaged for 1 n. Cell numbers were counted using the particle analyzer plugin of ImageJ. Data were analyzed using one-way ANOVA and Tukey's multiple comparison test. n = 17 (no light), n = 25 (light). C Cryosections were stained with DAPI and Iba-1. D 5–10 images of the central retina were taken of each eye and averaged for 1 n. Cell numbers were counted using the multi-point tool of ImageJ. Data were statistically analyzed using the Mann-Whitney U test. n = 3 (no light control), n = 6 (light + vehicle), n = 7 (light + IFN-β). B, D Graphs were plotted with GraphPad Prism 7.04. Bars represent mean ± SEM. SR: subretinal space; OPL: outer plexiform layer; RPE: retinal pigment epithelium; ONL: outer nuclear layer; INL: inner nuclear layer; GCL: ganglion cell layer.
Fig. 4
Fig. 4
Retinal thickness analysis using SD-OCT. Mice were treated with IFN-β (10,000 U) or PBS as vehicle every other day starting 1 h prior to light damage until 4 d afterwards. A Representative scans and B corresponding heatmaps compiled by spectral-domain optical coherence tomography (SD-OCT) displaying a retinal overview. C Retinal thickness was plotted with GraphPad Prism 7.04. Bars represent Mean ± SEM. Data were analyzed using one-way ANOVA and Tukey's multiple comparison test. n = 55 (no light control), n = 57 (light + vehicle), n = 63 (light + IFN-β).
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References

    1. Fritsche L.G., Igl W., Bailey J.N.C., Grassmann F., Sengupta S., Bragg-Gresham J.L., Burdon K.P., Hebbring S.J., Wen C., Gorski M., Kim I.K., Cho D., Zack D., Souied E., Scholl H.P.N., Bala E., Lee K.E., Hunter D.J., Sardell R.J., Mitchell P., Merriam J.E., Cipriani V., Hoffman J.D., Schick T., Lechanteur Y.T.E., Guymer R.H., Johnson M.P., Jiang Y., Stanton C.M., Buitendijk G.H.S., Zhan X., Kwong A.M., Boleda A., Brooks M., Gieser L., Ratnapriya R., Branham K.E., Foerster J.R., Heckenlively J.R., Othman M.I., Vote B.J., Liang H.H., Souzeau E., McAllister I.L., Isaacs T., Hall J., Lake S., Mackey D.A., Constable I.J., Craig J.E., Kitchner T.E., Yang Z., Su Z., Luo H., Chen D., Ouyang H., Flagg K., Lin D., Mao G., Ferreyra H., Stark K., von Strachwitz C.N., Wolf A., Brandl C., Rudolph G., Olden M., Morrison M.A., Morgan D.J., Schu M., Ahn J., Silvestri G., Tsironi E.E., Park K.H., Farrer L.A., Orlin A., Brucker A., Li M., Curcio C.A., Mohand-Saïd S., Sahel J.-A., Audo I., Benchaboune M., Cree A.J., Rennie C.A., V Goverdhan S., Grunin M., Hagbi-Levi S., Campochiaro P., Katsanis N., Holz F.G., Blond F., Blanché H., Deleuze J.-F., Igo R.P., Truitt B., Peachey N.S., Meuer S.M., Myers C.E., Moore E.L., Klein R., Hauser M.A., Postel E.A., Courtenay M.D., Schwartz S.G., Kovach J.L., Scott W.K., Liew G., Tan A.G., Gopinath B., Merriam J.C., Smith R.T., Khan J.C., Shahid H., Moore A.T., McGrath J.A., Laux R., Brantley M.A., Agarwal A., Ersoy L., Caramoy A., Langmann T., Saksens N.T.M., de Jong E.K., Hoyng C.B., Cain M.S., Richardson A.J., Martin T.M., Blangero J., Weeks D.E., Dhillon B., van Duijn C.M., Doheny K.F., Romm J., Klaver C.C.W., Hayward C., Gorin M.B., Klein M.L., Baird P.N., den Hollander A.I., Fauser S., Yates J.R.W., Allikmets R., Wang J.J., Schaumberg D.A., Klein B.E.K., Hagstrom S.A., Chowers I., Lotery A.J., Léveillard T., Zhang K., Brilliant M.H., Hewitt A.W., Swaroop A., Chew E.Y., Pericak-Vance M.A., DeAngelis M., Stambolian D., Haines J.L., Iyengar S.K., Weber B.H.F., Abecasis G.R., Heid I.M. A large genome-wide association study of age-related macular degeneration highlights contributions of rare and common variants, Nat. Genet. 2016;48:134–143. doi: 10.1038/ng.3448. - DOI - PMC - PubMed
    1. Klein R.J., Zeiss C., Chew E.Y., Tsai J.Y., Sackler R.S., Haynes C., Henning A.K., SanGiovanni J.P., Mane S.M., Mayne S.T., Bracken M.B., Ferris F.L., Ott J., Barnstable C., Hoh J. Complement factor H polymorphism in age-related macular degeneration. Science (80) 2005;308:385–389. doi: 10.1126/science.1109557. - DOI - PMC - PubMed
    1. Priller J., Flügel A., Wehner T., Boentert M., Haas C.A., Prinz M., Fernández-Klett F., Prass K., Bechmann I., de Boer B.A., Frotscher M., Kreutzberg G.W., Persons D.A., Dirnagl U. Targeting gene-modified hematopoietic cells to the central nervous system: use of green fluorescent protein uncovers microglial engraftment. Nat. Med. 2001;7:1356–1361. doi: 10.1038/nm1201-1356. - DOI - PubMed
    1. Karlstetter M., Scholz R., Rutar M., Wong W.T., Provis J.M., Langmann T. Retinal microglia: just bystander or target for therapy? Prog. Retin. Eye Res. 2015;45:30–57. doi: 10.1016/j.preteyeres.2014.11.004. - DOI - PubMed
    1. Karlstetter M., Langmann T. Adv. Exp. Med. Biol. Springer; New York, NY: 2014. Microglia in the aging retina; pp. 207–212. - DOI - PubMed

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