重组SPLUNC1蛋白对感染绵羊肺炎支原体的盘羊杂交羊的免疫调节作用研究

The immune regulation effect of the recombinant splunc1 protein in argali hybrid sheep infected with Mycoplasma ovipneumoniae

为研究重组SPLUNC1蛋白对感染绵羊肺炎支原体(MO)后的盘羊杂交羊免疫调节作用,本研究将6只巴什拜羊分为A组,18只盘羊杂交羊随机分为B、C、D组,对实验羊人工感染MO后第5 d,C组气管注射巴什拜羊重组SPLUNC1蛋白、D组气管注射盘羊杂交羊重组SPLUNC1蛋白,每天150μg/只;A、B组每天气管注射等量的生理盐水作为对照组。采用ELISA方法检测各组羊血清中MO抗体水平以及IL-8、IL-12、IL-13表达水平。结果显示,感染前所有实验羊MO抗体均为阴性,感染后第14 d所有实验羊MO抗体均为阳性;IL-8水平在感染后第14 d^21 d,A、C、D组极显著和显著地低于B组(p<0.01;p<0.05;p<0.01)。IL-12水平在感染后第14 d,A、C和D组极显著和显著地低于B组(p<0.01;p<0.05;p<0.01),第21 d时,A、C和D组极显著低于B组(p<0.01)。IL-13水平在感染后第5 d,A组显著低于B、C和D组(p<0.05),在第7 d^21 d,A、C、D组显著和极显著低于B组(p<0.05;p<0.01;p<0.01)。实验数据表明重组SPLUNC1蛋白对盘羊杂交羊具有明显的免疫调节作用。本研究为绵羊支原体肺炎的治疗提供了实验依据。

To investigate the immunomodulatory effects of the recombinant SPLUNC1 protein （rSPLUNC1） in argali hybrid sheep （AHS） infected with Mycoplasma ovipneumoniae （MO）, six Bashibay sheep （BS） as control （Group A）, eighteen AHS were equally divided into three groups of B, C and D. All the sheep were artificially infected with MO. On the 5th days post infection （DPI）, injected daily with rSPLUNC1 at a dose of 150μg （BS rSPLUNC1 for Group C and AHS rSPLUNC1 for Group D） or saline for Group A and B. The antibody level of MO and the levels of IL-8, IL-12, IL-13 in serum were detected by ELISA. The results showed that MO antibodies were negative in all sheep before infection, and MO antibodies were positive in all sheep at 14 DPI, indicating that artificial infection of MO was successful. The level of IL-8 in Group A, C and D were significantly lower than that in Group B （p〈0.01; p〈0.05; p〈0.01） during 14 to 21 DPI. In addition, the level of IL-12 in Group A, C and D were significantly lower than that in Group B （p〈0.01; p〈0.05; p〈0.01） at 14 DPI. The level of IL-12 in Group A, C and D were significantly lower than that in Group B （p〈0.01） at 21 DPI. However, the level of IL-13 in Group A were significantly lower than that in group B, C and D （p〈0.05） at 5 DPI. The level of IL-13 in Group A, C and D were significantly lower than that in Group B （p〈0.05; p〈0.01; p〈0.01） during 7 to 21 DPI. In conclusion, there were some definite immunomodulatory effects on AHS with the treatment of rSPLUNC1 to enhance the resistant to the MO, which provided a basis for further study on the treatment of MO.