Chen X, Threlkeld S W, Cummings E E, Sadowska G B, Lim Y-P, Padbury J F, Sharma S, Stonestreet B S
Department of Pediatrics, The Alpert Medical School of Brown University, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, RI 02905, USA.
J Comp Pathol. 2013 Feb;148(2-3):252-8. doi: 10.1016/j.jcpa.2012.06.001. Epub 2012 Jul 20.
Cytokines have gained increasing attention as therapeutic targets in inflammation-related disorders and inflammatory conditions have been investigated in sheep. Monoclonal antibodies (mAbs) specific for the ovine pro-inflammatory cytokines interleukin (IL)-1β and IL-6 could be used to study the effects of blocking pro-inflammatory cytokines in sheep. Ovine-specific IL-1β and IL-6 proteins and mAbs specific for these molecules were produced and the ability of the mAbs to neutralize the proteins was tested in cultures of ovine splenic mononuclear cells. Expression of nuclear factor (NF)-κβ and signal transducer and activator of transcription (STAT)-3 was evaluated by western blotting and densitometric quantification. Treatment with purified IL-1β and IL-6 proteins increased NF-κβ (P < 0.001) and STAT-3 (P < 0.01) expression, respectively, in cell culture. Treatment with these proteins that were pre-incubated with IL-1β and IL-6 mAbs attenuated (P < 0.01) these effects. These results confirm the bioactivity of ovine IL-1β and IL-6 proteins and the neutralizing capacity of anti-ovine-IL-1β and -IL-6 mAbs in vitro. These mAbs could be used to investigate anti-inflammatory strategies for attenuation of the effects of these pro-inflammatory cytokines in sheep.
细胞因子作为炎症相关疾病的治疗靶点受到越来越多的关注，并且已经在绵羊身上对炎症性疾病进行了研究。针对绵羊促炎细胞因子白细胞介素（IL）-1β和IL-6的单克隆抗体（mAb）可用于研究在绵羊中阻断促炎细胞因子的作用。制备了绵羊特异性的IL-1β和IL-6蛋白以及针对这些分子的mAb，并在绵羊脾单核细胞培养物中测试了mAb中和这些蛋白的能力。通过蛋白质印迹法和光密度定量法评估核因子（NF）-κβ和信号转导子及转录激活子（STAT）-3的表达。在细胞培养中，用纯化的IL-1β和IL-6蛋白处理分别增加了NF-κβ（P < 0.001）和STAT-3（P < 0.01）的表达。用预先与IL-1β和IL-6 mAb孵育的这些蛋白处理减弱了（P < 0.01）这些作用。这些结果证实了绵羊IL-1β和IL-6蛋白的生物活性以及抗绵羊IL-1β和-IL-6 mAb在体外的中和能力。这些mAb可用于研究减轻这些促炎细胞因子在绵羊中作用的抗炎策略。
