新疆野生一枝蒿粗多糖作为佐剂的免疫调节活性研究

Immunomodulatory activity of wild Artemisia rupestris L.crude polysaccharide as an adjuvant

目的探讨新疆野生一枝蒿粗多糖(WARCP)作为卵清白蛋白(OVA)疫苗佐剂肌肉免疫小鼠的增强效果。方法将ICR小鼠按随机数字表法分为6组(每组5只):9 g/L NaCl组(空白对照组)、OVA组(10μg OVA)、低剂量WARCP/OVA组(OVA+50μg WARCP)、中剂量WARCP/OVA组(OVA+200μg WARCP)、高剂量WARCP/OVA组(OVA+400μg WARCP)、铝佐剂/OVA组(阳性对照组,OVA+100μg Alum)。肌肉注射免疫小鼠,免疫后称量小鼠体质量,采用酶联免疫吸附测定法检测小鼠血清中的IgG及IgG1和IgG2a抗体水平,流式细胞术检测小鼠脾脏和淋巴结中的T细胞亚群。结果中剂量WARCP/OVA组可增强IgG、IgG1水平(均P<0.05),提高早期抗体水平;中剂量WARCP/OVA组和高剂量WARCP/OVA组可明显增强IgG2a水平(均P<0.05),且与Alum/OVA组相比差异均无统计学意义(均P>0.05)。低剂量WARCP/OVA组可提高小鼠脾脏CD4^+T细胞和淋巴结中CD4^+T、CD8^+T、CD4^+CD44^+T细胞比例;中剂量WARCP/OVA组和高剂量WARCP/OVA组可提高小鼠脾脏CD4^+T、CD8^+T、CD4^+CD44^+T、CD8^+CD44^+T细胞和淋巴结中CD8^+CD44^+T细胞比例,差异均具有统计学意义(均P<0.05);且WARCP免疫对小鼠体质量(均P>0.05)和生长无影响。结论植物来源的WARCP作为OVA蛋白疫苗佐剂可增强细胞免疫和体液免疫,且安全可靠,为WARCP的推广应用提供了理论依据。

Objective To investigate the enhancement effect of Xinjiang wild Artemisia rupestris L.crude polysaccharides(WARCP)as an adjuvant on ovalbumin(OVA)vaccine in mice immunized intramuscularly.Methods ICR mice were randomly divided into 6 groups(5 per group),including 9 g/L NaCl group(blank control),OVA group(10μg OVA),low dose WARCP/OVA group(OVA+50μg WARCP),medium dose WARCP/OVA group(OVA+200μg WARCP),high dose WARCP/OVA group(OVA+400μg WARCP),and aluminum adjuvant(Alum)/OVA group(positive control group,OVA+100μg Alum).ICR mice were immunized intramuscularly and weighted.The OVA-specific antibodies and subtypes in serum were detected by enzyme linked immunosorbent assay(ELISA).T cells subsets from spleen and lymph nodes were detected by flow cytometry.Results The medium-dose WARCP/OVA group enhanced IgG and IgG1 levels and increased early antibody levels(all P<0.05).The medium-dose WARCP/OVA group and the high-dose WARCP/OVA group significantly enhanced IgG2a levels(all P<0.05),but the difference was not statistically significant comparing with Alum/OVA group(P>0.05).The low-dose WARCP/OVA group enhanced the percentage of CD4^+T cells in spleen and CD4^+T,CD8^+T,CD4^+CD44^+T cells in lymph nodes(all P<0.05).The medium dose WARCP/OVA group and the high dose WARCP/OVA group enhanced the CD4^+T,CD8^+T,CD4^+CD44^+T,CD8^+CD44^+T cells in spleen and CD8^+CD44^+T cell in lymph nodes(all P<0.05).Conclusions Plant-derived WARCP as an OVA protein vaccine adjuvant can enhance cellular immunity and humoral immunity,and it is safe and reliable.The results in this study provide a theoretical basis for the popularization and application of WARCP.