Targeting of viral interleukin-10 with an antibody fragment specific to damaged arthritic cartilage improves its therapeutic potency.
Targeting of viral interleukin-10 with an antibody fragment specific to damaged arthritic cartilage improves its therapeutic potency.
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DOI：
10.1186/ar4613
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发表时间：
2014-07-16
影响因子：
4.9
通讯作者：
Nissim A
中科院分区：
文献类型：
--
作者：
Hughes C;Sette A;Seed M;D'Acquisto F;Manzo A;Vincent TL;Lim NH;Nissim A
我们之前证明，通过活性氧（ROS）翻译后修饰的 II 型胶原蛋白（CII）特异性的单链片段变量（scFv）可用于专门针对发炎关节炎关节的抗炎治疗。本研究的目的是在关节炎小鼠模型中证明抗 ROS 修饰的 CII（抗 ROS-CII）scFv 靶向发炎的关节炎关节时抗炎细胞因子的卓越功效。将病毒白细胞介素 10 (vIL-10) 通过基质金属蛋白酶 (MMP) 可裂解接头与抗 ROS-CII scFv (1-11E) 融合，形成 1-11E/vIL-10 融合体。通过酶联免疫吸附测定 (ELISA)、蛋白质印迹和关节炎软骨免疫染色测定 1-11E/vIL-10 与 ROS-CII 的结合，而 vIL-10 生物活性则通过使用 MC-9 细胞增殖测定进行体外评估。在抗原诱导的关节炎小鼠模型中测试了 1-11E/vIL-10 的特异性体内定位和治疗效果。 1-11E/vIL-10 特异性结合 ROS-CII 和受损的关节炎软骨。有趣的是，仅在用 MMP-1 裂解后才观察到融合蛋白的体外 vIL-10 活性。当对患有关节炎的小鼠进行全身给药时，1-11E/vIL-10 特异性地定位于患有关节炎的膝盖，并在 3 天后观察到峰值积累。此外，1-11E/vIL-10 比与对照抗鸡蛋溶菌酶 scFv (C7/vIL10) 融合的 vIL-10 更快地减少炎症。抗炎细胞因子的靶向递送增强了它们在关节炎小鼠模型中的抗关节炎作用。我们的结果进一步支持这样的假设：针对关节炎关节的生物治疗药物可以扩展到包括全身给药时缺乏功效的抗炎细胞因子。
We previously demonstrated that a single-chain fragment variable (scFv) specific to collagen type II (CII) posttranslationally modified by reactive oxygen species (ROS) can be used to target anti-inflammatory therapeutics specifically to inflamed arthritic joints. The objective of the present study was to demonstrate the superior efficacy of anti-inflammatory cytokines when targeted to inflamed arthritic joints by the anti-ROS modified CII (anti-ROS-CII) scFv in a mouse model of arthritis. Viral interleukin-10 (vIL-10) was fused to anti-ROS-CII scFv (1-11E) with a matrix-metalloproteinase (MMP) cleavable linker to create 1-11E/vIL-10 fusion. Binding of 1-11E/vIL-10 to ROS-CII was determined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and immune-staining of arthritic cartilage, whereas vIL-10 bioactivity was evaluated in vitro by using an MC-9 cell-proliferation assay. Specific in vivo localization and therapeutic efficacy of 1-11E/vIL-10 was tested in the mouse model of antigen-induced arthritis. 1-11E/vIL-10 bound specifically to ROS-CII and to damaged arthritic cartilage. Interestingly, the in vitro vIL-10 activity in the fusion protein was observed only after cleavage with MMP-1. When systemically administered to arthritic mice, 1-11E/vIL-10 localized specifically to the arthritic knee, with peak accumulation observed after 3 days. Moreover, 1-11E/vIL-10 reduced inflammation significantly quicker than vIL-10 fused to the control anti-hen egg lysozyme scFv (C7/vIL10). Targeted delivery of anti-inflammatory cytokines potentiates their anti-arthritic action in a mouse model of arthritis. Our results further support the hypothesis that targeting biotherapeutics to arthritic joints may be extended to include anti-inflammatory cytokines that lack efficacy when administered systemically.
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影响因子：
27.4
作者：
van Holten, J;Pavelka, K;Tak, PP
通讯作者：
Tak, PP
影响因子：
5.4
作者：
Gu, Yongpeng;Yang, Jianfei;Ouyang, Xinshou;Liu, Weicheng;Li, Hongxing;Yang, Jianjun;Bromberg, Jonathan;Chen, Shu-Hsia;Mayer, Lloyd;Unkeless, Jay C.;Xiong, Huabao
通讯作者：
Xiong, Huabao
影响因子：
--
作者：
Joosten, LAB;Lubberts, E;vandenBerg, WB
通讯作者：
vandenBerg, WB
影响因子：
13.3
作者：
Lim, Ngee Han;Meinjohanns, Ernst;Nagase, Hideaki
通讯作者：
Nagase, Hideaki
影响因子：
15.3
作者：
Ding, Y;Qin, L;Kotenko, S V;Pestka, S;Bromberg, J S
通讯作者：
Bromberg, J S