免疫刺激DNA序列与过敏原联用对哮喘小鼠模型气道过敏性炎症的作用

Modulation of allergic airway inflammation by immunostimulatory DNA sequences in conjunction with an allergen in a murine model of asthma

目的 观察免疫刺激DNA序列 (ISS DNA)单用和与过敏原卵白蛋白 (OVA)合用 ,对支气管哮喘 (简称哮喘 )小鼠模型气道过敏性炎症的作用及作用维持时间。方法 BALB/c小鼠 36只 ,分ISS组 (A组 ) 12只、ISS +OVA组 (B组 ) 12只、OVA组 (C组 ) 6只和生理盐水 (NS)组 (D组 ) 6只。A、B、C组用OVA致敏及激发。A组和B组根据注射次数再分A1、B11次注射组 (1次腹腔注射ISS DNA 10 0μg或ISS DNA 10 0 μg +OVA 10 μg)和A2 、B2 2次注射组 (2次腹腔注射ISS DNA或ISS DNA +OVA)。各组在第 1次激发后 6周中 ,每周采血 1次测特异性IgE ,最后处死小鼠测支气管肺泡灌洗液 (BALF)中细胞计数及分类 ,肺组织病理检查 ,检测脾细胞分泌γ干扰素 (IFN γ)的水平。结果 BALF中嗜酸细胞 (EOS)数A1组为 (2 39± 0 81)× 10 4/ml;A2 组为 (2 .6 2± 0 .77)× 10 4/ml;B1组为 (1.80± 0 .12 )× 10 4/ml;B2 组为 (1.84± 0 .6 7)× 10 4/ml,与C组 [(12 .4 3± 2 .13)× 10 4/ml]比较差异有显著性 (P <0 .0 5 )。脾细胞分泌的IFN γA1组为 (5 10± 10 2 )pg/ml;A2 组为 (492± 98)pg/ml;B1组为 (5 32± 12 0 )pg/ml;B2组为 (46 9± 132 )pg/ml,与C组 [(194± 80 )pg/ml]比较差异有显著性 (P <0 .0 5 )。血清IgE前

Objective To compare the suppressive effects of immunostimulatory seqences ( ISS DNA) alone or ISS DNA in conjunction with ovalbumin (OVA) on airway inflammation in a mouse model of asthma. Methods Thirty six female BALB/c mice were divided into 4 groups: group ISS(A), group ISS+OVA(B), group OVA (C) and group normal saline (D). Mice in groups A, B and C were sensitized and challenged with OVA. Group A and group B were subdivided into subgroup A 1 and B 1 (injected intraperitoneally with ISS DNA 100 μg or ISS DNA 100 μg and OVA 10 μg once) and subgroup A 2 and B 2 (injected intraperitoneally with ISS DNA or ISS DNA and OVA twice). Blood samples were obtained every week for six weeks. OVA specific IgE was measured by ELISA. Bronchoalveolar lavage fluid (BALF), lung tissues and spleen cells were collected . BAL total cell numbers and differentials were counted. Interferonγ(IFN γ) produced by OVA stimulated spleen cells was determined by ELISA. Results Eosinophil count in group A 1 [(2.39±0.81)×10 4/ml], group A 2 [(2.62±0.77)×10 4/ml], group B 1 [(1.80±0.12)×10 4/ml], and group B 2 [(1.84±0.67)×10 4/ml] were significantly lower than those in group C [(12.43±2.13)×10 4/ml], P <0.05. The levels of IFN γ in group A 1 [(510±102) pg/ml], group A 2 [(492±98) pg/ml],group B 1 [(532±120) pg/ml], and group B 2 [(469±132) pg/ml] were significantly higher than those in group C [(194±80) pg/ml], P <0.05. The serum level of IgE in group B was significantly lower than that in group C in four weeks, but that in group A was not significantly differentfrom that in group C. A second dose of ISS DNA did not show additional effect as compared to the signal dose treatment.Conclusions ISS DNA inhibited allergic airway inflammation in a murine model of asthma. ISS DNA and OVA combination was more effective than ISS DNA alone.