Regulation by Immune Response to Vaccine by Botanicals

推动这个项目的假设是，今天广泛使用的一些植物性药物可以起到 口服免疫调节剂，或皮下注射免疫佐剂与疫苗混合，增强或抑制对联合注射疫苗的免疫反应。在我们的经验中，最好的佐剂是QS-21和GPI-0100三萜皂苷，这些皂苷来自南美树的树皮。然而，迫切需要更有效的方法来进一步增强疫苗的免疫原性。当使用Globo H-KLH和GD2-KLH结合疫苗时，将确定所选植物药物的佐剂和免疫调节剂活性。 Globo H是一种六糖类自身抗原，在乳腺癌和大多数其他上皮性癌症中过度表达。GD2是一种神经节苷脂自身抗原，在肉瘤、神经母细胞瘤和黑色素瘤中过度表达。锁孔帽血蓝蛋白(KLH)是从锁孔帽血中提纯的一种有效的免疫载体蛋白。对Globo H和KLH的抗体反应将通过酶联免疫吸附试验、流式细胞仪和细胞毒性试验来测定，T细胞对KLH的反应将通过增殖和ELISPOT试验来测定。这些植物药将在目标1中作为单独佐剂与Globo H-KLH疫苗混合进行测试，在目标2中作为免疫调节剂进行测试 口服，在AIM 3中作为佐剂与GPI-0100联合使用，或在AIM 4中使用GD2-KLH疫苗加植物作为佐剂或免疫调节剂在已确定的GD2阳性癌症的背景下使用。在任何化验中被发现持续有效的植物药将被转回研究资源核心进行进一步的纯化和表征。所得纯化组分的佐剂和免疫调节活性将再次进行测试。这样，广泛使用的具有佐剂或免疫调节活性的植物药将被识别出来，并确定其最有效的成分。虽然我们的偏见是，精制组分或精制组分的组合将出现峰值反应性，但也有可能出现较不纯的植物混合物的峰值反应性，因为可能存在我们目前不了解的操作问题和因素。此外，关于复杂佐剂配方的效力也有很长的历史记录。在我们寻找免疫活性最强的植物或植物部分时，我们将警惕这种可能性。我们还将警惕我们筛选的植物药物中的负面免疫调节活性，因为这将表明不应与疫苗一起服用或在其他重要的新免疫反应时服用植物药物。因此，我们有两个议程贯穿每个具体目标：1)确定广泛使用的植物药物对免疫系统识别和对抗免疫原的能力的积极或消极影响，以及2)确定这些植物药物中最有效的部分或成分，作为佐剂或作为免疫调节剂与抗癌疫苗一起使用。

The hypothesis driving this project is that some of the botanicals in wide use today can act as immunomodulators when taken orally, or immunological adjuvants when administered subcutaneously mixed with vaccines, augmenting or dampening the immune response against the co-administered vaccine. The optimal adjuvants in our experience are the QS-21 and GPI-0100 triterpenoid saponins derived from the bark of the South American tree Quillaja saponaria Molina. There is, however, a pressing need for more potent approaches to further augmenting vaccine immunogenicity. Adjuvant and immunomodulator activity will be determined for selected botanicals when administered with globo H-KLH and GD2-KLH conjugate vaccines. Globo H is a hexaccharide auto-antigen overexpressed in breast cancers and most other epithelial cancers. GD2 is a ganglioside auto-antigen overexpressed on sarcomas, neuroblastomas, and melanomas. Keyhole limpet hemocyanin (KLH) is a potent immunological carrier protein purified from the blood of the keyhole limpet. The antibody response to globo H and KLH will be determined in ELISA, FACS, and cytotoxicity assays, and the T-cell response to KLH will be measured in proliferation and ELISPOT assays. The botanicals will be tested mixed with the globo H-KLH vaccine in Aim 1 as adjuvant alone, in Aim 2 as immunomodulator administered orally, in Aim 3 as adjuvant in combination with GPI-0100, or in Aim 4 with a GD2-KLH vaccine plus botanical as adjuvant or immunomodulator in the setting of established, GD2 positive cancer. Botanicals found to be consistently active in any assay will be referred back to the Research Resource Core for further purification and characterization. Adjuvant and immunomodulator activity of the resulting purified components will be tested again. In this way, widely used botanicals with adjuvant or immunomodulator activity will be identified and their most active components defined. While our bias is that peak reactivity will occur with purified fractions or combinations of purified fractions, it is also possible that peak reactivity will occur with less pure botanical mixtures since there may be issues and factors operating that we do not currently understand. Also, there is a long history documenting the potency of complex adjuvant formulations. In our search for the most immunoreactive botanicals or botanical fractions, we will be alert to this possibility. We will also be alert for negative immunomodulatory activity in the botanicals we screen, as this will suggest botanicals that should not be taken with vaccines or at other times when new immune responses are important. We have, therefore, 2 agendas that run through each of the specific aims: 1) to determine the positive or negative impact of widely used botanicals on the immune system's ability to recognize and react against immunogens, and 2) to define the most active fractions or components of these botanicals for use as adjuvants or as immunomodulators with vaccines against cancer.