STAT3-Specific Single Domain Nanobody Inhibits Expansion of Pathogenic Th17 Responses and Suppresses Uveitis in Mice.
STAT3-Specific Single Domain Nanobody Inhibits Expansion of Pathogenic Th17 Responses and Suppresses Uveitis in Mice.
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DOI：
10.3389/fimmu.2021.724609
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发表时间：
2021
影响因子：
7.3
通讯作者：
Egwuagu CE
中科院分区：
文献类型：
--
作者：
Mbanefo EC;Yan M;Kang M;Alhakeem SA;Jittayasothorn Y;Yu CR;Parihar A;Singh S;Egwuagu CE
STAT3 激活调节哺乳动物细胞生长、分化和存活的基因转录。 T 细胞中 Stat3 的基因缺失已被证明可以消除 Th17 分化，这表明 STAT3 是 Th17 介导的疾病的潜在治疗靶点。然而，细胞内蛋白（例如 STAT3）的治疗靶向的主要障碍是缺乏将 STAT3 抑制剂递送到细胞中的有效方法。在这项研究中，我们开发了一种由 STAT3 特异性重链分子的可变 (V) 区组成的新型抗体 (SBT-100)，并证明这种 15 kDa STAT3 特异性纳米抗体进入人和小鼠细胞，并以浓度依赖性方式诱导抑制 STAT3 活化和淋巴细胞增殖。为了研究 SBT-100 是否能有效抑制体内炎症，我们通过使用眼部自身抗原、光感受器间视黄醇结合蛋白 (IRBP651-670) 的肽进行主动免疫，在 C57BL/6J 小鼠中诱导实验性自身免疫性葡萄膜炎 (EAU)。通过眼底镜检查、组织学检查或光学相干断层扫描对视网膜进行的分析表明，用 SBT-100 治疗小鼠可通过抑制介导 EAU 的致病性 Th17 细胞的扩增来抑制葡萄膜炎。暗适应和光适应 a 波和 b 波的视网膜电图 (ERG) 记录表明，SBT-100 治疗可以使小鼠避免出现在未治疗的 EAU 小鼠中观察到的严重视力障碍。来自未经治疗的 EAU 小鼠的活化 IRBP 特异性 T 细胞的过继转移诱导了 EAU，而在接受来自 SBT-100 治疗小鼠的 IRBP 特异性 T 细胞的小鼠中，EAU 显着减弱。总而言之，这些结果证明了 SBT-100 对小鼠的功效，并表明其治疗人类自身免疫性疾病的潜力。
STAT3 activates transcription of genes that regulate cell growth, differentiation, and survival of mammalian cells. Genetic deletion of Stat3 in T cells has been shown to abrogate Th17 differentiation, suggesting that STAT3 is a potential therapeutic target for Th17-mediated diseases. However, a major impediment to therapeutic targeting of intracellular proteins such as STAT3 is the lack of efficient methods for delivering STAT3 inhibitors into cells. In this study, we developed a novel antibody (SBT-100) comprised of the variable (V) region of a STAT3-specific heavy chain molecule and demonstrate that this 15 kDa STAT3-specific nanobody enters human and mouse cells, and induced suppression of STAT3 activation and lymphocyte proliferation in a concentration-dependent manner. To investigate whether SBT-100 would be effective in suppressing inflammation in vivo, we induced experimental autoimmune uveitis (EAU) in C57BL/6J mice by active immunization with peptide from the ocular autoantigen, interphotoreceptor retinoid binding protein (IRBP651-670). Analysis of the retina by fundoscopy, histological examination, or optical coherence tomography showed that treatment of the mice with SBT-100 suppressed uveitis by inhibiting expansion of pathogenic Th17 cells that mediate EAU. Electroretinographic (ERG) recordings of dark and light adapted a- and b-waves showed that SBT-100 treatment rescued mice from developing significant visual impairment observed in untreated EAU mice. Adoptive transfer of activated IRBP-specific T cells from untreated EAU mice induced EAU, while EAU was significantly attenuated in mice that received IRBP-specific T cells from SBT-100 treated mice. Taken together, these results demonstrate efficacy of SBT-100 in mice and suggests its therapeutic potential for human autoimmune diseases.