Topical ocular treatment with monoclonal antibody Fab fragments targeting Japanese cedar pollen Cry j 1 inhibits Japanese cedar pollen-induced allergic conjunctivitis in mice
Topical ocular treatment with monoclonal antibody Fab fragments targeting Japanese cedar pollen Cry j 1 inhibits Japanese cedar pollen-induced allergic conjunctivitis in mice
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使用针对日本柳杉花粉 Cry j 1 的单克隆抗体 Fab 片段局部眼部治疗可抑制日本柳杉花粉诱导的小鼠过敏性结膜炎
DOI：
10.1016/j.ejphar.2017.01.028
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Shin Yoshino
中科院分区：
文献类型：
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作者：
Nobuaki Mizutani;Takeshi Nabe;Shin Yoshino
由于缺少Fc部分，抗体的Fab片段（Fab）只能与特定的过敏原结合，因此缺乏效应功能。在本研究中，我们检测了针对杉木花粉（JCP） Cry j 1的IgG1单克隆抗体（mAb） fab是否能够调节JCP诱导的小鼠变应性结膜炎。用局部给药JCP滴眼液反复刺激对JCP敏感的BALB/c小鼠。用木瓜蛋白酶消化抗JCP IgG1单抗（P1-3和P1-8）制备的fab在JCP攻击前15分钟应用于眼睛，然后测量临床结膜炎评分。在体外实验中，P1-3和P1-8与JCP Cry j1有特异性结合。此外，完整的P1-3与Cry j 1的结合被P1-3 fab抑制，但P1-8 fab不受抑制；此外，P1-8 fab抑制了P1-8的完整结合，而P1-3 fab则没有，这表明P1-3和P1-8识别的Cry j 1表位不同。局部用P1-3或P1-8 Fabs治疗后，jcp诱导的结膜炎明显受到抑制（P<0.01）。组织学评价P1-8 fab显示结膜内嗜酸性粒细胞浸润减少（P<0.01）。这些结果表明，局部用IgG1 mAb fab治疗Cry j 1可有效抑制jcp诱导的小鼠变应性结膜炎。此外，这表明fab识别的某些表位可能作为调节过敏性结膜炎的工具。
Fab fragments (Fabs) of antibodies having the ability only to bind to specific allergens lack effector functions due to the absence of the Fc portion. In the present study, we examined whether IgG1 monoclonal antibody (mAb) Fabs targeting Japanese cedar pollen (JCP) Cry j 1 were able to regulate JCP-induced allergic conjunctivitis in mice. BALB/c mice actively sensitized with JCP were repeatedly challenged by topical administration of JCP eye drops. Fabs prepared by the digestion of anti-JCP IgG1 mAbs (P1-3 and P1-8) with papain were applied to the eye 15 min before the JCP challenges followed by measurement of the clinical conjunctivitis score. In the in vitro experiments, P1-3 and P1-8 showed specific binding to JCP Cry j 1. Furthermore, intact P1-3 binding to Cry j 1 was inhibited by P1-3 Fabs, but not P1-8 Fabs; additionally, P1-8 Fabs, but not P1-3 Fabs, suppressed the intact P1-8 binding, suggesting that the epitopes of Cry j 1 recognized by P1-3 and P1-8 were different. Topical ocular treatment with P1-3 Fabs or P1-8 Fabs was followed by marked suppression of JCP-induced conjunctivitis (P<0.01). In histological evaluation, P1-8 Fabs showed a reduction in eosinophil infiltration in the conjunctiva (P<0.01). These results demonstrated that topical ocular treatment with IgG1 mAb Fabs to Cry j 1 was effective in suppressing JCP-induced allergic conjunctivitis in mice. Furthermore, it suggests the possibility that some epitopes recognized by Fabs could be used as a tool to regulate allergic conjunctivitis.