Nagai T, Suzuki Y, Kiyohara H, Susa E, Kato T, Nagamine T, Hagiwara Y, Tamura S, Yabe T, Aizawa C, Yamada H
Oriental Medicine Research Center, The Kitasato Institute, 5-9-1 Shirokane, Minato-ku, 108-8642, Tokyo, Japan.
Vaccine. 2001 Sep 14;19(32):4824-34. doi: 10.1016/s0264-410x(01)00215-8.
Active substances from hot water extracts from 267 different Chinese and Japanese medicinal herbs were screened for mucosal adjuvant activity with influenza HA vaccine in mice. The extract from the root of Polygala tenuifolia was found to contain potent mucosal adjuvant activity. The active substances were purified and identified as onjisaponins A, E, F, and G. When each onjisaponin (10 microg) was intranasally (i.n.) inoculated with influenza vaccine (10 microg) in mice, serum hemagglutination-inhibiting (HI) antibody titers increased 3-14 times over control mice administered vaccine alone after 4 weeks. When each onjisaponin (10 microg) was i.n. inoculated with the vaccine (10 microg) followed by i.n. vaccination of the vaccine alone after 3 weeks, serum HI antibody titers increased 27-50 fold over those mice given i.n. vaccinations without onjisaponins. These same conditions also significantly increased nasal anti-influenza virus IgA antibody titers. Two inoculations with onjisaponin F (1 microg) and influenza HA vaccine (1 microg) at 3 weeks intervals, significantly increased serum HI antibody and nasal anti-influenza virus IgA and IgG antibody titers after only 1 week over mice given HA vaccine alone after the secondary vaccination. Intranasal vaccination with onjisaponin F inhibited proliferation of mouse adapted influenza virus A/PR/8/34 in bronchoalveolar lavages of infected mice. Separate intranasal vaccinations with onjisaponins A, E, F, and G (10 microg) each and diphtheria-pertussis-tetanus (DPT) vaccine (10 microg) of mice followed by i.n. vaccination with DPT vaccine alone after 4 weeks showed significant increases in serum IgG and nasal IgA antibody titers after 2 weeks following secondary vaccination over mice vaccinated with DPT vaccine alone. All onjisaponins showed little hemolytic activity at concentrations up to 100 microg/ml. The results of this study suggest that onjisaponins may provide safe and potent adjuvants for intranasal inoculation of influenza HA and DPT vaccines.
对267种不同的中日药草热水提取物中的活性物质进行了筛选，以检测其对小鼠流感血凝素（HA）疫苗的黏膜佐剂活性。发现远志根提取物具有强大的黏膜佐剂活性。活性物质被纯化并鉴定为远志皂苷A、E、F和G。当将每种远志皂苷（10微克）与流感疫苗（10微克）经鼻接种到小鼠体内时，4周后血清血凝抑制（HI）抗体滴度比仅接种疫苗的对照小鼠提高了3至14倍。当将每种远志皂苷（10微克）经鼻接种疫苗（10微克），3周后再单独经鼻接种疫苗时，血清HI抗体滴度比未接种远志皂苷经鼻接种疫苗的小鼠提高了27至50倍。相同条件下也显著提高了鼻内抗流感病毒IgA抗体滴度。间隔3周两次接种远志皂苷F（1微克）和流感HA疫苗（1微克），仅1周后血清HI抗体以及鼻内抗流感病毒IgA和IgG抗体滴度就比二次接种后仅接种HA疫苗的小鼠显著提高。经鼻接种远志皂苷F可抑制小鼠适应株流感病毒A/PR/8/34在感染小鼠支气管肺泡灌洗液中的增殖。分别将远志皂苷A、E、F和G（10微克）经鼻接种到小鼠体内，再接种白喉-百日咳-破伤风（DPT）疫苗（10微克），4周后再单独经鼻接种DPT疫苗，二次接种2周后血清IgG和鼻内IgA抗体滴度比仅接种DPT疫苗的小鼠显著提高。所有远志皂苷在浓度高达100微克/毫升时溶血活性都很低。本研究结果表明，远志皂苷可为经鼻接种流感HA疫苗和DPT疫苗提供安全有效的佐剂。
