CAJ | 학술논문

背景白细胞介素17(IL-17)是一种促炎性细胞因子,在多种器官移植免疫排斥反应和自身免疫性疾病中发挥致病作用.IL-17拮抗剂在器官移植排斥反应中的作用被广泛研究,但其对角膜移植后排斥反应防治作用的研究较少. 目的探讨抗小鼠IL-17单克隆抗体对小鼠角膜移植排斥反应的抑制效果.方法选取8 ～10周龄C57 BL/6小鼠25只和BALB/c小鼠50只,将直径2 mm的C57BL/6小鼠双眼中央角膜作为供体角膜移植到50只BALB/c小鼠右眼角膜植床上,11-0尼龙线间断缝合8针,建立小鼠角膜移植模型.将建立的40只角膜移植小鼠模型按随机数字表法随机分为2个组,术后腹腔内分别注射小鼠IL-17中和性抗体(IL-17中和性抗体组)或同型对照抗体(同型对照抗体组),并于术后不同时间点根据Plskova等的标准对受体角膜植片的炎症反应情况进行评分,≥5分为发生排斥反应.应用免疫荧光组织化学法检测受体植片炎性细胞的浸润；用逆转录PCR法对植片中的炎性细胞进行定量；应用ELISA法分析两组受体小鼠Th1、Th2和Th17相关细胞因子在脾脏中的表达情况.结果与同型对照抗体组相比,IL-17中和性抗体可以提高角膜植片的存活率(P＜0.05).IL-17中和性抗体组受体植片中性粒细胞、CD4+T淋巴细胞和CD8+T淋巴细胞mRNA含量分别为2.22±0.10、1.64±0.04、1.32±0.10,明显低于同型对照抗体组的3.61±0.08、2.69±0.06、2.17±0.04,差异均有统计学意义(P=0.000、0.000、0.000).同型对照抗体组受体小鼠脾脏中γ-干扰素(IFN-γ)、IL-12 p40和IL-17质量浓度分别为(741.48±10.51)、(1156.90±69.93)、(366.13 ±7.93) ng/L,明显高于IL-17中和性抗体组的(529.80±13.83)、(539.58±10.74)、(173.70±8.11) ng/L,差异均有统计学意义(P=0.000、0.001、0.000).结论中和IL-17的生物学活性可以在一定程度上抑制同种异体角膜移植术后的免疫排斥反应.
배경 백세포개소17(IL-17)시일충촉염성세포인자,재다충기관이식면역배척반응화자신면역성질병중발휘치병작용.IL-17길항제재기관이식배척반응중적작용피엄범연구,단기대각막이식후배척반응방치작용적연구교소. 목적 탐토항소서IL-17단극륭항체대소서각막이식배척반응적억제효과.방법 선취8 ～10주령C57 BL/6소서25지화BALB/c소서50지,장직경2 mm적C57BL/6소서쌍안중앙각막작위공체각막이식도50지BALB/c소서우안각막식상상,11-0니룡선간단봉합8침,건립소서각막이식모형.장건립적40지각막이식소서모형안수궤수자표법수궤분위2개조,술후복강내분별주사소서IL-17중화성항체(IL-17중화성항체조)혹동형대조항체(동형대조항체조),병우술후불동시간점근거Plskova등적표준대수체각막식편적염증반응정황진행평분,≥5분위발생배척반응.응용면역형광조직화학법검측수체식편염성세포적침윤；용역전록PCR법대식편중적염성세포진행정량；응용ELISA법분석량조수체소서Th1、Th2화Th17상관세포인자재비장중적표체정황.결과 여동형대조항체조상비,IL-17중화성항체가이제고각막식편적존활솔(P＜0.05).IL-17중화성항체조수체식편중성립세포、CD4+T림파세포화CD8+T림파세포mRNA함량분별위2.22±0.10、1.64±0.04、1.32±0.10,명현저우동형대조항체조적3.61±0.08、2.69±0.06、2.17±0.04,차이균유통계학의의(P=0.000、0.000、0.000).동형대조항체조수체소서비장중γ-간우소(IFN-γ)、IL-12 p40화IL-17질량농도분별위(741.48±10.51)、(1156.90±69.93)、(366.13 ±7.93) ng/L,명현고우IL-17중화성항체조적(529.80±13.83)、(539.58±10.74)、(173.70±8.11) ng/L,차이균유통계학의의(P=0.000、0.001、0.000).결론 중화IL-17적생물학활성가이재일정정도상억제동충이체각막이식술후적면역배척반응.
Background Interleukin-17 (IL-17)is a potent pro-inflammatory cytokine and plays a pathogenic role in autoimmune disease.It was confirmed that IL-17 is implicated in allograft rejection of many transplanted organs.Recent studies have foensed on the effect of IL-17 antagonists on allograft rejection.Objective This study aimed to investigate the inhibitory effect of anti-mouse IL-17 monoclonal antibody (mAb) on corneal allograft rejection.Methods Twenty-five 8 to 10-week-old C57BL/6 mice and 50 BALB/c mice were collected.Donor cornea grafts with 2 mm diameter from 25 C57BL/6 mice was transplanted to 50 eye of BALB/c mice to establish a model of corneal transplantation.The recipients were randomized into 2 groups,and neutralizing mouse IL-17antibody or isotype control antibody was intraperitoneally injected immediately after transplantation for experimental treatment,respectively.Allografts were scored clinically at appropriate time points after treatment based on Plskova criteria,and ≥5 was confirmed as rejection.Infiltrating cells in corneal graft were detected qualitatively and quantitatively by immunohistochemistry and reverse transcription-PCR separately.The cytokine levels of T helper type 1 (Th1),Th2,and Th17 in recipients' spleen wer(c) analyzcd by ELISA.The use of the animals followed the Statement of ARVO.Results Compared with the isotype control antibody group,the survival of grafts was improved in the IL-17mAb group(P＜0.05).The levels of neutrophile granulocyte mRNA,CD4+ and CD8+ T lymphotes mRNA were 2.22±0.10,1.64±0.04 and 1.32±0.10 in the IL-17 mAb group,showing a significant decline in comparison with those of the isotype control antibody group(3.61 ±0.08,2.69±0.06 and 2.17±0.04) (P=0.000,0.000,0.000).Interferon-γ(IFN-γ),IL-12 p40 and IL-17 concentrations in recipients ' splenocytes were (529.80 ± 13.83) ng/L,(539.58 ±10.74) ng/L and(173.70±8.11)ng/L in the IL-17 mAb group,and thosc in the isotype control antibody group were (741.48± 10.51) ng/L,(1156.90 ± 69.93) ng/L and (366.13± 7.93) ng/L,with significant differences between them (P=0.000,0.001,0.000).Conclusions Neutralization IL-17 bioactivity inhibits mouse corneal allograft rejection to a certain extent.