抗细胞间黏附分子-1单克隆抗体对大鼠实验性血管痉挛的抑制作用

Inhibition of experimental vasospasm with anti-intercellular adhesion molecule-1 monoclonal antibody in rats.

作者信息

Oshiro E M, Hoffman P A, Dietsch G N, Watts M C, Pardoll D M, Tamargo R J

机构信息

Department of Neurological Surgery, Johns Hopkins University School of Medicine, Baltimore, Md, USA.

出版信息

Stroke. 1997 Oct;28(10):2031-7; discussion 2037-8. doi: 10.1161/01.str.28.10.2031.

DOI:10.1161/01.str.28.10.2031

PMID:9341715

Abstract

BACKGROUND AND PURPOSE

Inflammation may play a role in delayed chronic vasospasm after aneurysmal subarachnoid hemorrhage. We investigated the role of intercellular adhesion molecule-1 (ICAM-1) and macrophage/granulocyte infiltration in the rat femoral artery model of vasospasm using systemic administration of a murine anti-ICAM-1 monoclonal antibody (MAb).

METHODS

The femoral arteries (n = 72) in Sprague-Dawley rats (n = 36) were enclosed in latex pouches bilaterally. Autologous blood was injected into the pouch on one side, and saline was injected on the contralateral side. Chronic vessel narrowing was evaluated with the use of 29 rats, which were randomized into one of three groups for intraperitoneal injections: (1) anti-ICAM-1 MAb (2 mg/kg per dose, n = 10), (2) isotype-matched MAb (2 mg/kg per dose, n = 9), or (3) saline (n = 10), given at 3 hours and 3, 6, and 9 days after blood exposure. These rats were killed 12 days after blood exposure, and femoral artery lumen cross-sectional areas were determined by computerized image analysis. Saturation of ICAM-1 binding sites with this dosing schedule was evaluated by fluorescence-activated cell sorter (FACS) analysis of splenocytes. Immunohistochemical studies with objective cell counts were performed to evaluate macrophage/granulocyte infiltration at 24 hours in 7 rats, comparing anti-ICAM-1 MAb treatment (n = 4) with isotype-matched control MAb (n = 3).

RESULTS

Animals treated with anti-ICAM-1 MAb showed a significant inhibition of arterial narrowing at 12 days (P = .0081), with lumen patency of 96.5 +/- 5.3% (mean +/- SEM), compared with 77.3 +/- 5.6% for isotype-matched MAb and 72.2 +/- 5.3% for saline-treated controls. FACS analysis of splenocytes from animals treated with anti-ICAM-1 MAb confirmed saturation of ICAM-1 binding sites. Vessels treated with anti-ICAM-1 MAb showed a significant decrease in inflammatory cell infiltrates, with objective macrophage/granulocyte counts of 31.3 +/- 26.6 (mean +/- SEM) per high-powered field, compared with 171.4 +/- 30.7 for isotype-matched control MAb (P = .0027).

CONCLUSIONS

Anti-ICAM-1 MAb administered systemically starting 3 hours after blood exposure results in significant inhibition of chronic vasospasm in the rat femoral artery model and is correlated with a reduction in the number of infiltrating macrophages and granulocytes in the periadventitial region of blood-exposed arteries. We conclude that inflammatory changes associated with ICAM-1-mediated macrophage and granulocyte migration play an important role in the development of posthemorrhagic chronic vasospasm in this model.

摘要

背景与目的

炎症可能在动脉瘤性蛛网膜下腔出血后的迟发性慢性血管痉挛中起作用。我们使用鼠抗细胞间黏附分子-1（ICAM-1）单克隆抗体（MAb）全身给药，在大鼠股动脉血管痉挛模型中研究了ICAM-1和巨噬细胞/粒细胞浸润的作用。

方法

将36只Sprague-Dawley大鼠双侧股动脉（n = 72）置于乳胶袋中。在一侧袋中注入自体血，对侧注入生理盐水。对29只大鼠进行慢性血管狭窄评估，将其随机分为三组进行腹腔注射：（1）抗ICAM-1 MAb（每剂量2 mg/kg，n = 10），（2）同型匹配MAb（每剂量2 mg/kg，n = 9），或（3）生理盐水（n = 10），在血液暴露后3小时及3、6和9天给药。这些大鼠在血液暴露后12天处死，并通过计算机图像分析确定股动脉管腔横截面积。通过对脾细胞的荧光激活细胞分选仪（FACS）分析评估该给药方案下ICAM-1结合位点的饱和度。对7只大鼠在24小时时进行免疫组织化学研究并进行客观细胞计数，比较抗ICAM-1 MAb治疗组（n = 4）和同型匹配对照MAb组（n = 3）的巨噬细胞/粒细胞浸润情况。

结果

抗ICAM-1 MAb治疗的动物在12天时动脉狭窄得到显著抑制（P = 0.0081），管腔通畅率为96.5±5.3%（平均值±标准误），而同型匹配MAb组为77.3±5.6%，生理盐水处理对照组为72.2±5.3%。对抗ICAM-1 MAb治疗动物的脾细胞进行FACS分析证实了ICAM-1结合位点的饱和。抗ICAM-1 MAb处理的血管炎症细胞浸润显著减少，每个高倍视野的客观巨噬细胞/粒细胞计数为31.3±26.6（平均值±标准误），而同型匹配对照MAb组为171.4±30.7（P = 0.0027）。