SAP基因注射干预小鼠SLE样综合征的发生

SAP gene injection interfere the occurrence of murine SLE-like syndrome

目的 :研究SAP基因注射对SLE发病的干预作用 ,并进一步探讨SAP发挥作用的可能机制。方法 :通过RT PCR方法克隆SAP基因 ,构建其真核表达质粒pcDNA3 SAP ,观察SAP基因注射对活化淋巴细胞免疫诱导小鼠产生的SLE样综合征的干预作用 ,以ELISA方法检测抗dsDNA抗体的产生情况 ,以免疫荧光法检测肾脏免疫复合物沉积 ;通过巨噬细胞吞噬实验观察SAP与DNA结合后对DNA清除的影响 ;用增殖实验检测巨噬细胞吞噬SAP结合的DNA后对预致敏淋巴细胞活化的影响。结果 :SAP基因注射可有效干预小鼠SLE样综合征的发生 ,SAP与活化淋巴细胞DNA结合后可明显促进巨噬细胞对DNA的吞噬 ,并且巨噬细胞吞噬SAP结合的DNA后不引起预致敏淋巴细胞的增殖。结论

Objective:To study the interference effect of the SAP gene injection on the SLE-like syndrome and to discuss its possible mechanism.Methods:The target DNA encoding SAP was cloned by RT-PCR, and its eukaryotic expression vector-pcDNA3-SAP was constructed, then observed the interference effect of the SAP gene injection on the SLE-like syndrome induced by activated lymphocytes immunization, the anti-dsDNA antibodies were detected by ELISA and immune complexes in glomeruli were observed by immunofluorescent stain. The clearance of DNA was observed by phagocytosis of macrophages and the proliferation assay was used to detect the influence of macrophages' ingesting SAP-binding DNA on the activation of primed lymphocytes.Results:SAP gene inoculation could interfere the occurrence of SLE-like syndrome, the binding of SAP to DNA from activated lymphocytes significantly promoted the uptake of DNA by macrophages, and macrophages that ingested the SAP-binding DNA would not cause the increased proliferation of primed lymphocytes. Conclusion:It suggested that SAP could interfere the occurrence of SLE through promoting the efficient clearance of autoantigenic DNA.