抑制素基因免疫大鼠的免疫应答与基因表达

Immune Response and Gene Expression in Rats after Inhibin Gene Immunization

为了研究抑制素基因免疫对大鼠免疫应答和发情的影响及免疫后抑制素的组织分布,将60只大鼠分为5组,每只分别肌肉注射10(T1)、50(T2)、100μg pCIS(T3),50μg pcDNA3.1(V)和生理盐水(S)。ELISA检测抑制素抗体水平,阴道涂片法检测大鼠的发情状况,免疫组化分析抑制素的组织分布。结果显示,不同剂量抑制素基因2次免疫10 d后抗体P/N值均显著升高(P<0.05),3次免疫10 d后T2和T3组抗体P/N值进一步显著升高(P<0.05),T3组抗体水平有高于T1和T2组的趋势(P>0.05);抑制素基因免疫对大鼠的发情无显著影响;3次免疫2周后心脏、肝脏、接种肌肉部位均未检出抑制素;卵巢、肾脏和垂体部位均检出抑制素;而脾脏部位,抑制素质粒免疫组检出抑制素,对照组则未检出抑制素。这些结果表明,免疫剂量和次数的增加没有导致大鼠产生明显的抑制素免疫耐受,抑制素基因免疫大鼠是相对安全的,抑制素的组织分布结果亦为抑制素基因免疫的作用机理研究提供了理论依据。

To study the immune response, estrus and tissue distribution of inhibin after inhibin gene immunization, 60 rats were divided into five groups and immunized （i. m. ） with 10（T1） ,50 （T2）,100 μg pCIS（T3）, 50 μg pcDNA3.1（V）and 0.85%NaCl （S） respectively. The antibody against inhibin was detected by ELISA, the estrus was detected by vagina smear and the tissue distribution of inhibin was detected by immunohistochemistry. The results showed that the P/N value of inhibin antibody went up significantly at dayl0 after 1^st booster（P〈0.05）and the P/N value in T2 and T3 group raised significantly at day 20 after 2^nd booster（P〈0.05）. The P/N value in T3 group was slightly higher than those in the T1 and T2 group（P〉0.05）. The estrus was not affected. The immunohistochemistry results showed that there was no positive staining in the tissues such as heart, liver and muscles at inoculated sites, however, positive staining occurred in ovary, kidney and pituitary for both the experimental and control groups 2 weeks after 2^nd booster. In the spleen, positive staining appeared in the experimental groups, but not in the control groups. These results suggested that the increased immunization time and dosage of inhibin DNA did not cause significant immunologic tolerance. The inhibin gene is safe to immunize rats and the tissue distribution of inhibin also provides important evidence to elucidate the mechanism of gene immunization.