纳米铝佐剂吸附HBsAg及其免疫学效应的研究

纳米材料用作疫苗佐剂,已受到极大重视1～4].Kreuter等5]于1981年首次将纳米材料应用于疫苗佐剂.但直到20世纪90年代初,纳米生物学领域才有了长足进步6].铝佐剂包括Al(OH)3和AlPO4两种,而Al(OH)3是目前唯一被FDA认证的人用佐剂.本文自制纳米铝佐剂,通过物理吸附HBsAg,以常规铝佐剂为对照,研究Balb/c小鼠和豚鼠对两种佐剂疫苗的体液免疫应答和细胞免疫应答.1实验部分1.1材料、仪器和实验动物HBsAg纯抗原、OPD、辣根过氧化物酶标记的羊抗Balb/c小鼠IgG、ConA、RPMI1640、小牛血清、3H-TdR、氢氧化铝佐剂商业品(2mg/mL);纳米…

Immune Effect of HBsAg Adsorbed by Nanoparticulate Alum adjuvant

We have synthesized a novel formulation of nanoparticulate(NP) alum adjuvant specifically in the cationic water-in-oil microemulsions of water/benzalkonium bromide(BB), octyl alcohol/cyclohexane at 30 ℃, which was characterized to be crystalline Al(OH)3 with average particle diameter about 70 nm by the analysis of X-ray diffraction and to have a spherical shape structure by TEM photography analysis. Then adsorption degree was assayed. The OD492 of the NP is about 10 times higher than the bulk(BK) alum adjuvant. Compared to bulk alum adjuvant, we studied the immune effect of the NP alum adjuvant with HBsAg vaccine, which was injected intraperitoneally into Guinea pig, and serum antibody titers of the first and second week after immunization were higher in NP group than BK group by ELISA(P<0.01, P<0.05); Groin immuno-injection intraperitoneally into Balb/c mice, was performed once or there times, prolife responses of spleen cells were determined by 3H thymidine incorporation; non-specific phagocytosis of macrophage of abdominal cavity was measured by lack of color Malachite Green and IL-2 was assayed by activation mouse spleen cells. It is indicated that all cellular immunity bioassay results were higher in NP group than in BK group, except non-specific phagocytosis of macrophage of abdominal cavity by there times immunity. However, there was no significant differerces. Thus, nanoparticulate alum adjuvant can be employed as a more effective adjuvant for HBsAg vaccine than bulk alum adjuvant in the earlier period of post-immunity.