目的探讨脊髓损伤(SCI)处局部给予LINGO-1多克隆抗体治疗的可行性。方法24只成年雌性SD大鼠采用随机数字表法分为假手术组、半横断对照IgG组及半横断LINGO-1多克隆抗体组,每组8只。假手术组仅行椎板切除术,另外两组均行T9脊髓半横断术,半横断LINGO-1多克隆抗体组大鼠半横断损伤后立刻通过微量渗透泵局部给予损伤处LINGO-1多克隆抗体,而半横断对照IgG组仅给予兔源性对照IgG。术后3d和28d取各组T8-10节段脊髓制作冰冻切片,应用免疫荧光染色方法观察分析LINGO-1多克隆抗体是否进入脊髓组织并与LINGO-1分子特异性结合。结果半横断LINGO-1多克隆抗体组大鼠SCI术后3d和28d均可检测出兔源性抗体;术后3d,半横断对照IgG组切片LINGO-1染色强度(1.704±0.174)明显强于半横断LINGO-1多克隆抗体组(0.568±0.052),LINGO-1多克隆抗体预处理后的半横断对照IgG组切片LINGO-1染色强度(0.329±0.055)明显弱于半横断对照IgG组(1.704±0.174)切片,比较差异均有统计学意义(P〈0.05)。结论局部给予的LINGO-1多克隆抗体在较宽的时间窗里均可进入SCI区并特异性识别LINGO-1分子.证明被动免疫治疗SCI是可行的。
Objective To analyze the feasibility of local LINGO-1 polyclonal antibody administration for treatment of spinal cord injury in adult rats. Methods Twenty-four adult female SD rats were randomized into sham-operated group, rabbit IgG group and LINGO-1 antibody group. In the latter two groups, partial transaction of the T9 segment of the spinal cord was performed to completely sever the dorsal corticospinal tract, followed immediately by administration of rabbit IgG and anti-LINGO polyclonal antibody via a mini-osmotic pump, respectively. At 3 and 28 days after the operation, the T8-10 segments of the spinal cord were harvested to prepare cryosections, and immunofluorescence staining was used to analyze the penetration of LINGO-1 polyclonal antibody into the spinal cord tissue and its specific binding to LINGO-1 molecules. Results In LINGO-1 antibody group, the presence of rabbit antibodies was detected at the injured sites of the spinal cord at 3 and 28 days after the operation. The mean immunofluorescenee density was significantly lower in LINGO-1 antibody group than in rabbit IgG group at 3 days after the operation (P〈0.05). In rabbit IgG group, the mean immunofluorescence density for L1NGO-1 in the crysections pre-treated with LINGO-1 polyclonal antibody was significantly lower than that in sections pre-treated with rabbit IgG(P〈0.05). Conclusion Locally administered LINGO-1 polyclonal antibody can penetrate into the injured sites in the spinal cord in a wide time window and recognizes LINGO-1 molecule specifically, suggesting the feasibility of passive immunotherapy for spinal cord injury.