Kukhetpitakwong Ratiya, Hahnvajanawong Chariya, Homchampa Preecha, Leelavatcharamas Vichai, Satra Jarunee, Khunkitti Watcharee
Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
Int Immunopharmacol. 2006 Nov;6(11):1729-35. doi: 10.1016/j.intimp.2006.07.008. Epub 2006 Aug 4.
Pods of Acacia concinna (Leguminosae) contain several saponins. In this study, four saponin fractions which were acetone fraction (AAC), aqueous fraction (WAC), hydromethanolic fraction (HAC) and methanolic fraction (MAC) were generated and their haemolytic activities and surface activities were determined in comparison with quillaja saponin (QS). There were no significant differences between the haemolytic activities of MAC and QS. However, the surface tensions of MAC was significantly lower than QS (p < 0.001). Furthermore, the immunomodulatory effect and the adjuvant potential of MAC on the cellular and humoral immune response of BALB/c mice against ovalbumin were investigated. The splenocyte proliferations induced by MAC were significantly higher than QS at the concentrations of 200, 400, 800 and 1000 microg/ml (p < 0.05). BALB/c mice were immunized subcutaneously either with OVA 20 microg alone or with OVA 20 microg combining with QS (10 microg) or MAC (10 and 40 microg). Ten days after the second immunization, concanavalin A (Con A)-, pokeweed mitogen (PWM)-, and OVA-stimulated splenocyte proliferation and OVA-specific antibodies in serum were measured. The results suggested that MAC (40 microg) could activate T and B cells. In addition, OVA-specific IgG, IgG1 IgG2a and IgG2b antibody levels in serum were significantly enhanced by MAC (40 microg) as compared with OVA control group (p < 0.001). This finding suggested that MAC might be effect on Th1 and Th2 helper T cells. In conclusion, the results indicated that MAC at a dose of 40 microg could be used as vaccine adjuvant to increase immune responses.
儿茶(豆科)的豆荚中含有多种皂苷。在本研究中,制备了四个皂苷级分,分别是丙酮级分(AAC)、水级分(WAC)、氢甲醇级分(HAC)和甲醇级分(MAC),并与皂树皂苷(QS)比较,测定了它们的溶血活性和表面活性。MAC和QS的溶血活性之间没有显著差异。然而,MAC的表面张力显著低于QS(p<0.001)。此外,还研究了MAC对BALB/c小鼠针对卵清蛋白的细胞免疫和体液免疫反应的免疫调节作用及佐剂潜力。在浓度为200、400、800和1000μg/ml时,MAC诱导的脾细胞增殖显著高于QS(p<0.05)。BALB/c小鼠皮下注射单独的20μg卵清蛋白(OVA),或与10μg QS或10μg和40μg MAC联合的20μg OVA。第二次免疫后10天,测定了伴刀豆球蛋白A(Con A)、商陆有丝分裂原(PWM)和OVA刺激的脾细胞增殖以及血清中的OVA特异性抗体。结果表明,40μg的MAC可激活T细胞和B细胞。此外,与OVA对照组相比,40μg的MAC显著提高了血清中OVA特异性IgG、IgG1、IgG2a和IgG2b抗体水平(p<0.001)。这一发现表明MAC可能对Th1和Th2辅助性T细胞有作用。总之,结果表明40μg剂量的MAC可作为疫苗佐剂以增强免疫反应。