重组红火蚁毒素蛋白SoliⅣ致病机理的初步研究

Pathogenetic Study of the Recombinant Solenopsis Invicta Buren Venom Allergen Soli Ⅳ

作者通过研究重组红火蚁毒素蛋白Solenopsis invicta Ⅳ(Soli Ⅳ)对兔及外周血淋巴细胞的影响,探讨红火蚁毒素蛋白致病机制。通过分离兔外周血淋巴细胞进行培养,经不同浓度的重组红火蚁毒素蛋白Soli Ⅳ分别和细菌脂多糖(LPS)、刀豆蛋白(ConA)共同刺激后,MTT法测定淋巴细胞的增殖情况;体内试验:用重组蛋白Soli Ⅳ从兔背部皮下注入,观察兔的临床反应,定期采血,用ELISA方法测定兔血清中白介素-4(IL-4)和总IgE的变化。重组红火蚁毒素蛋白Soli Ⅳ浓度为25、50、75μg/ml和LPS共刺激时,与单独LPS刺激对照组相比,淋巴细胞增殖活性显著增高(P<0.05);浓度为15、25、50、75μg/ml和ConA共刺激时,与单独ConA刺激对照组比较,淋巴细胞增殖活性显著增高(P<0.05);重组Soli Ⅳ过敏兔的血清中IL-4和总IgE的水平升高,在20 h左右达到最高值。试验结果表明,一定浓度的重组红火蚁毒素蛋白Soli Ⅳ能引起体外培养的T、B淋巴细胞增殖,过敏体质兔在重组红火蚁毒素蛋白Soli Ⅳ刺激后能引起Ⅰ型变态反应。

The pathogenesis of Solenopsis invicta Buren venom allergen soli IV was analyzed by studying the effects of Solenopsis invicta IV （Soli IV） on rabbit and lymphocytic function of rabbit peripheral blood. After expression, separation and purification, the recombination protein Soli Ⅳwas used to stimulate lymphocyte of rabbit alone or combined with LPS and ConA in vitro, and the lymphocyte proliferation was measured by the method of MTT assay. The clinic reaction was recorded and the sera level of IL-4 and total IgE was analyzed using enzyme linked imunosorbent assay （ELISA） after the rabbits were injected with soll Ⅳsubcutaneously. Combined with LPS, 25 μg/ml, 50 μg/ml, 75 μg/ml of recombinant Soli IV treatment could promote lymphocyte proliferation of peripheral blood significantly. When combination with ConA, 15 μg/ml, 25 μg/ml, 50 μg/ ml and 75 μg/ml of recombination SoliⅣ could promote significantly lymphocyte proliferation of peripheral blood. The sera level of IL-4 and total IgE rose to the top at about 20 h after stimulation. Soli IV could promote significantly T and B lymphocyte proliferation in vitro at a certain concentration, and hypersusceptibility type I would be reduced in rabbit of hypersusceptible habitus.