Crystallin β-b2 promotes retinal ganglion cell protection in experimental autoimmune uveoretinitis

IF 4.2 3区 医学 Q2 NEUROSCIENCES
Dirk Bauer, Michael R. R. Böhm, Xiaoyu Wu, Bo Wang, Tida Viola Jalilvand, Martin Busch, Maren Kasper, Katrin Brockhaus, Lena Wildschütz, Harutyun Melkonyan, Björn Laffer, Gerd Meyer Zu Hörste, Arnd Heiligenhaus, Solon Thanos
{"title":"Crystallin β-b2 promotes retinal ganglion cell protection in experimental autoimmune uveoretinitis","authors":"Dirk Bauer, Michael R. R. Böhm, Xiaoyu Wu, Bo Wang, Tida Viola Jalilvand, Martin Busch, Maren Kasper, Katrin Brockhaus, Lena Wildschütz, Harutyun Melkonyan, Björn Laffer, Gerd Meyer Zu Hörste, Arnd Heiligenhaus, Solon Thanos","doi":"10.3389/fncel.2024.1379540","DOIUrl":null,"url":null,"abstract":"Crystallin βb2 (crybb2) is upregulated in regenerating retinas and in various pathological conditions of the retina, including uveoretinitis. However, the role of crybb2 in this disease is largely unknown. Therefore, we used recombinant crybb2 (rcrybb2) as intravitreal treatment of B10.RIII mice prior to immunization with human interphotoreceptor retinoid-binding protein peptide 161–180 (hIRBPp161-180) in complete Freund’s adjuvant (CFA) and concomitant injection of pertussis toxin (PTX) to induce experimental autoimmune uveoretinitis (EAU). In naïve mice, more beta III-tubulin (TUBB3) + and RNA-binding protein with multiple splicing (RBPMS) + cells were found in the ganglion cell layer of the retina than in EAU eyes, suggesting a loss of retinal ganglion cells (RGC) during the development of EAU. At the same time, the number of glial fibrillary acidic protein (GFAP) + cells increased in EAU eyes. RGCs were better protected in EAU eyes treated with rcrybb2, while the number of GFAP+ cells decreased. However, in retinal flatmounts, both retinal ganglion cells and retinal endothelial cells stained positive for TUBB3, indicating that TUBB3 is present in naïve B10.RIII mouse eyes not exclusive to RGCs. A significant decline in the number of RBPMS-positive retinal ganglion cells was observed in retinal flatmounts from EAU retinas in comparison to naïve retinas or EAU retinas with intravitreal rcrybb2 treatment. Whereas no significant decrease in TUBB3 levels was detected using Western blot and RT-qPCR, GFAP level, as a marker for astrocytes, increased in EAU mice compared to naïve mice. Level of <jats:italic>Bax</jats:italic> and <jats:italic>Bcl2</jats:italic> in the retina was altered by treatment, suggesting better cell survival and inhibition of apoptosis. Furthermore, our histologic observations of the eyes showed no change in the incidence and severity of EAU, nor was the immune response affected by intravitreal rcrybb2 treatment. Taken together, these results suggest that intravitreal injection of rcrybb2 reduces retinal RGC death during the course of EAU, independent of local or systemic autoimmune responses. In the future, treating posterior uveitis with rcrybb2 to protect RGCs may offer a promising novel therapeutic strategy.","PeriodicalId":12432,"journal":{"name":"Frontiers in Cellular Neuroscience","volume":null,"pages":null},"PeriodicalIF":4.2000,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in Cellular Neuroscience","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3389/fncel.2024.1379540","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"NEUROSCIENCES","Score":null,"Total":0}
引用次数: 0

Abstract

Crystallin βb2 (crybb2) is upregulated in regenerating retinas and in various pathological conditions of the retina, including uveoretinitis. However, the role of crybb2 in this disease is largely unknown. Therefore, we used recombinant crybb2 (rcrybb2) as intravitreal treatment of B10.RIII mice prior to immunization with human interphotoreceptor retinoid-binding protein peptide 161–180 (hIRBPp161-180) in complete Freund’s adjuvant (CFA) and concomitant injection of pertussis toxin (PTX) to induce experimental autoimmune uveoretinitis (EAU). In naïve mice, more beta III-tubulin (TUBB3) + and RNA-binding protein with multiple splicing (RBPMS) + cells were found in the ganglion cell layer of the retina than in EAU eyes, suggesting a loss of retinal ganglion cells (RGC) during the development of EAU. At the same time, the number of glial fibrillary acidic protein (GFAP) + cells increased in EAU eyes. RGCs were better protected in EAU eyes treated with rcrybb2, while the number of GFAP+ cells decreased. However, in retinal flatmounts, both retinal ganglion cells and retinal endothelial cells stained positive for TUBB3, indicating that TUBB3 is present in naïve B10.RIII mouse eyes not exclusive to RGCs. A significant decline in the number of RBPMS-positive retinal ganglion cells was observed in retinal flatmounts from EAU retinas in comparison to naïve retinas or EAU retinas with intravitreal rcrybb2 treatment. Whereas no significant decrease in TUBB3 levels was detected using Western blot and RT-qPCR, GFAP level, as a marker for astrocytes, increased in EAU mice compared to naïve mice. Level of Bax and Bcl2 in the retina was altered by treatment, suggesting better cell survival and inhibition of apoptosis. Furthermore, our histologic observations of the eyes showed no change in the incidence and severity of EAU, nor was the immune response affected by intravitreal rcrybb2 treatment. Taken together, these results suggest that intravitreal injection of rcrybb2 reduces retinal RGC death during the course of EAU, independent of local or systemic autoimmune responses. In the future, treating posterior uveitis with rcrybb2 to protect RGCs may offer a promising novel therapeutic strategy.
晶体蛋白β-b2促进实验性自身免疫性葡萄膜视网膜炎视网膜神经节细胞的保护
晶体蛋白βb2(crybb2)在视网膜再生和视网膜的各种病理状态(包括葡萄膜视网膜炎)中上调。然而,crybb2 在这种疾病中的作用在很大程度上是未知的。因此,我们在用完全弗氏佐剂(CFA)中的人视网膜受体间结合蛋白肽161-180(hIRBPp161-180)免疫B10.RIII小鼠并同时注射百日咳毒素(PTX)诱导实验性自身免疫性葡萄膜视网膜炎(EAU)之前,使用重组crybb2(rcrybb2)对小鼠进行玻璃体内治疗。与EAU小鼠相比,在幼稚小鼠的视网膜神经节细胞层中发现了更多的βⅢ-tubulin(TUBB3)+细胞和具有多重剪接的RNA结合蛋白(RBPMS)+细胞,这表明在EAU的发展过程中视网膜神经节细胞(RGC)丢失了。与此同时,EAU眼球中胶质纤维酸性蛋白(GFAP)+细胞的数量有所增加。在接受rcrybb2治疗的EAU眼中,RGC得到了更好的保护,而GFAP+细胞的数量则减少了。然而,在视网膜平片上,视网膜神经节细胞和视网膜内皮细胞的TUBB3染色均呈阳性,这表明TUBB3存在于天真B10.RIII小鼠的眼球中,并不局限于RGC。在EAU视网膜平片中观察到,与未出生视网膜或接受玻璃体内rcrybb2治疗的EAU视网膜相比,RBPMS阳性视网膜神经节细胞的数量明显下降。虽然通过 Western 印迹和 RT-qPCR 检测到 TUBB3 的水平没有明显下降,但作为星形胶质细胞标记物的 GFAP 水平在 EAU 小鼠中比在幼稚小鼠中有所增加。视网膜中的 Bax 和 Bcl2 水平因治疗而改变,这表明细胞存活率提高,凋亡受到抑制。此外,我们对小鼠眼睛的组织学观察显示,玻璃体内注射 rcrybb2 并没有改变 EAU 的发病率和严重程度,也没有影响免疫反应。综上所述,这些结果表明,玻璃体内注射rcrybb2可减少EAU病程中视网膜RGC的死亡,而不受局部或全身自身免疫反应的影响。未来,用rcrybb2治疗后葡萄膜炎以保护RGC可能是一种很有前景的新型治疗策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
CiteScore
7.90
自引率
3.80%
发文量
627
审稿时长
6-12 weeks
期刊介绍: Frontiers in Cellular Neuroscience is a leading journal in its field, publishing rigorously peer-reviewed research that advances our understanding of the cellular mechanisms underlying cell function in the nervous system across all species. Specialty Chief Editors Egidio D‘Angelo at the University of Pavia and Christian Hansel at the University of Chicago are supported by an outstanding Editorial Board of international researchers. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信