过表达细胞因子信号传导抑制蛋白1的树突状细胞过继免疫对COPD模型中T细胞相关因子的影响

Effects of dendritic cell-based adoptive immunotherapy with over-expressed SOCS1 on Th17-and Tregrelated cytokines in the BALF of COPD mice

目的树突状细胞(DCs)、辅助性T细胞17(Th17)、调节性T细胞(Treg)与慢性阻塞性肺疾病(COPD)发病密切相关。文章观察过表达细胞因子信号传导抑制蛋白1(SOCS1)的DCs过继免疫小鼠COPD模型支气管肺泡灌洗液(BALF)中Th17、Treg相关细胞因子的变化,为COPD治疗提供新思路。方法 48只雄性C57BL/6小鼠按随机数字表法分成5组(除健康对照组外,其他各组均造模):健康对照组(暴露于空气中正常饲养)、模型组(烟熏造模第1天用等量等渗盐水0.5mL/只尾静脉注射)、imDCs 1×106组、DCs?SOCS1低浓度组、DCs?SOCS1高浓度组,其中imDCs组、DCs?SOCS1低浓度组、DCs?SOCS1高浓度组又分为第1天和第7天2个观察点,每个观察点6只小鼠。imDCs组分别于第1、7天尾静脉注射1×106个imDCs,4个DCs?SOCS1组分别于第1、7天尾静脉注射1×106、2×106个过表达SOCS1的DCs。取健康对照组、模型组肺组织作石蜡切片及HE染色;ELISA检测除健康对照组外各组BALF中Th17细胞因子白细胞介素?17(IL?17)、IL?23,Treg相关细胞因子IL?10、转换生长因子?β(TGF?β)的变化。结果与模型组第1天、第7天IL?17表达[(78.87±1.08)、(78.87±1.08)pg/mL]比较,imDCs组[(46.46±0.77)、(55.69±0.35) pg/mL]、DCs?SOCS1低浓度组[(34.09±3.98)、(35.65±0.54)pg/mL]、DCs?SOCS1高浓度组[(24.12±0.57)、(27.00±0.58)pg/mL]明显降低(P<0.05);与模型组第1天、第7天IL?23[(200.62±0.65)、(200.62±0.65)pg/mL]比较,imDCs[(150.19±0.53)、(167.70±1.73)pg/mL]、DCs?SOCS1低浓度组[(121.09±0. 53)、(136.34±0.90)pg/mL]、DCs?SOCS1高浓度组[(70.21±0.91)、(99.35±1.83)pg/mL]显著降低(P<0.05);与模型组第1天、第7天IL?10[(39.46±3.88)、(39.46±3.88)pg/mL]比较,imDCs[(50.74±1.77)、(44.56±2.63) pg/mL]、DCs?SOCS1 1×106[(58.71±3.84)、(54.78±1.43) pg/mL]、DCs?SOCS1高浓度组[(70.12±2.62)、(63.00±2.57) pg/mL]显著增高(P<0.05);与模型组第1天、第7天TGF?β[(24.98±0.43)、(24.98±0.43)pg/mL]比较,imDCs组[(36.46±0.98)、(33.27±0.92)pg/mL]、DCs?SOCS1低浓度组[(42.40±0.62)、(40.12±0.83)pg/mL]、DCs?SOCS1高浓度组[(50.55±0.53)、(44.98±0.52)pg/mL]显著增高(P<0.05)。组内比较,DCs?SOCS1低浓度组第7天较第1天IL?23含量增高,IL?10、TGF?β含量显著降低(P<0.05);DCs?SOCS1高浓度组第7天较第1天IL?17、IL?23含量增高,IL?10、TGF?β含量降低(P<0.05)。结论小鼠尾静脉注射SOCS1过表达的DCs后可抑制COPD中Th17细胞相关细胞因子分泌,效果优于单纯注射imDCs,且与浓度、时间有关。

Objective Dendritic cells(DCs),helper T cells 17(Th17) and regulatory T cells(Treg) are closely related to the pathogenesis of chronic obstructive pulmonary disease (COPD). This study aimed to investigate the changes of Th17- and Treg-related cytokines in the bronchoalveolar lavage fluid(BALF) of COPD mice after DC-based adoptive immunotherapy with over-expressed suppressor of cytokine signaling protein 1(SOCS1)and provide some new ideas for the treatment of COPD. Methods A total of 48 male C57BL/6 mice were randomly divided into 5 groups:healthy control,COPD model control,immature DC(imDC),DC-SOCS1 1×106,and DC-SOCS1 2×106. The healthy controls were exposed to air and fed normally,the COPD model controls injected with normal saline at 0.5 mL/ on the first day of modeling by fumigation,the mice of the imDC group injected via the tail vein with 1 ×106 imDCs,and those of the DC-SOCS1 groups injected with 1 ×106 or 2 ×106 DCs with over expressed SOCS1,all via the tail vein on the 1st and 7th day of modeling. Then the lung tissues were collected from the mice for preparation of paraffin sections and HE staining,and ELISA was employed for determination of the levels of Th17-related IL-17 and IL-23 and Treg- related IL-10 and TGF-β in the BALF of the model mice. Results Compared with the COPD model controls,the mice in the imDC,DC-SOCS1 1×106 and DC-SOCS1 2×106 groups showed significantly decreased levels of IL-17 on the 1st day([78.87 ± 1.08]vs[46.46 ± 0.77],[34.09 ± 3.98]and[24.12 ± 0.57]pg/mL,P < 0.05)and 7th day after modeling([78.87 ± 1.08]vs[55.69 ±0.35],[35.65 ± 0.54]and[27.00 ± 0.58]pg/mL,P < 0.05),and IL-23 on the 1st day([200.62 ± 0.65]vs[150.19 ± 0.53],[121.09 ± 0. 53]and[70.21 ± 0.91]pg/mL,P < 0.05)and 7th day([200.62 ± 0.65]vs[167.70 ± 1.73],[136.34 ± 0.90]and[99.35 ± 1.83]pg/mL,P < 0.05),but remarkably increased levels of IL-10 on the 1st day([39.46 ± 3.88]vs[50.74 ± 1.77],[58.71 ± 3.84]and[70.12 ± 2.62]pg/mL,P < 0.05)and 7th day([39.46 ± 3.88]vs[44.56 ± 2.63],[54.78 ± 1.43]and[63.00 ± 2.57]pg/mL,P < 0.05),TGF-β on the 1st day([24.98 ± 0.43]vs[36.46 ± 0.98],[42.40 ± 0.62]and [50.55 ± 0.53]pg/mL,P < 0.05)and 7th day([24.98 ± 0.43]vs[33.27 ± 0.92],[40.12 ± 0.83]and[44.98 ± 0.52]pg/mL,P < 0.05). The contents of IL-17 and IL-23 were markedly lower while those of IL-10 and TGF-β higher in the DC-SOCS1 1×106 than in the imDC group(P < 0.05),and the levels of the former two significantly higher and those of the latter two lower in the DC-SOCS1 2× 106 than in the DC-SOCS1 1×106 group(P < 0.05). Conclusion Transfusion of DCs with over-expressed SOCS1 can inhibit the secretion of Th17-related cytokines in COPD,and the effect is better than that of imDCs alone and related to the concentration and time.