Oxygen as a master immunologic switch

氧气作为主要免疫开关

• 批准号: 8183092
• 负责人: John R Ohlfest
• 金额: $ 31.33万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-09 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8183092
• 关键词: ANXA2 gene; Adjuvant; Affect; Age; Antibodies; Antibody Formation; Antigens; B-Lymphocytes; Binding; Biological Assay; Brain Neoplasms; Breast Carcinoma; CD8B1 gene; Cancer Etiology; Cell Culture Techniques; Cells; Cessation of life; Clinic; Clinical; Clinical Trials; Complex; Cross Presentation; Cross-Priming; Cultured Tumor Cells; Cytotoxic T-Lymphocytes; Data; Dendritic Cells; Enhancing Antibodies; Fc Receptor; Figs - dietary; Foundations; Glioblastoma; Glioma; Goals; Harvest; Human; Hypoxia; Hypoxia Inducible Factor; Immune response; Immunoglobulin G; Immunologics; Immunotherapy; Knowledge; Ligands; Malignant Glioma; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Methods; Modeling; Molecular; Morbidity - disease rate; Mus; Nervous System Neoplasms; Oxygen; Oxygen measurement, partial pressure, arterial; Patients; Property; Recombinants; Relative (related person); Research Personnel; Rest; Role; Site; Source; Survival Rate; System; T-Lymphocyte; TLR2 gene; Testing; Therapeutic; Toll-Like Receptor 2; Toxic effect; Treatment Efficacy; Vaccinated; Vaccination; Vaccine Antigen; Vaccine Production; Vaccines; Work; bHLH-PAS factor HLF; base; cancer cell; cancer immunotherapy; cell killing; cytokine; immunogenicity; improved; in vivo; loss of function; lymphocyte proliferation; mouse model; neoplastic cell; novel; novel vaccines; response; tissue culture; trafficking; translational approach; tumor; vaccine efficacy

DESCRIPTION (provided by applicant): Malignant brain tumors are the leading cause of cancer-related death in people under the age of 35. Immunotherapy in the form of personalized vaccines has demonstrated immunologic activity and clinical responses in select glioma patients with minimal toxicity. There are numerous ongoing clinical trials that utilize cultured tumor cells as the source of vaccine antigen for the treatment of a wide array of tumors. However, very little is known about how cell culture conditions affect the immune response and ultimate clinical response following vaccination. The consistent condition used is expansion of tumor cells in atmospheric oxygen (20% O2). We have identified the oxygen concentration used in tissue culture as a primary determinant of the immunogenicity of tumor cell vaccines. Our data have led to the attractive central hypothesis that the oxygen tension in a tumor cell culture acts as a master immunologic switch, dictating the type and strength of immune response induced by vaccination. We have reproducibly shown that lysates from glioma cells cultured in 5% O2 prime cytotoxic T lymphocytes (CTLs) with superior effector functions relative to lysate from glioma cells cultured in 20% O2 in human and murine systems. This difference profoundly affects the efficacy of immunotherapy, as shown by significant improvements in survival in murine models of glioma and breast carcinoma. We demonstrated that administration of 5% O2 lysate vaccination caused superior CTL proliferation, cytokine elaboration, tumoricidal function, and trafficking to tumor sites relative to 20% O2 lysate vaccines. Conversely, 20% O2 lysate vaccines enhance antibody responses. Despite reduced tumor-reactive antibody responses, the 5% O2 vaccines require B cells for therapeutic efficacy, revealing a putative role of B cells in CTL priming. Additionally, we have evidence that glioma cells grown in 5% O2 upregulate toll-like receptor (TLR) 2 ligands because TLR2 is required in several systems to distinguish the immunogenicity of 5% and 20% O2 lysates. The goal of this proposal is to elucidate the molecular basis of the "oxygen switch effect" in order to rationally improve the efficacy of tumor cell vaccines. In specific aim 1 we will determine and optimize tumor cell intrinsic changes that modulate immunogenicity. Pharmacologic strategies to increase hypoxia inducible factors (HIF) in tumor cells will be compared to actual hypoxia in cell cultures that will subsequently be assayed for immunogenicity. Additionally, we will test the hypothesis that HIF2a is the molecular switch that induces expression of TLR2 ligands. In specific aim 2, we will test our hypothesis that B cells are required because IgG-tumor lysate complexes trigger Fc receptor-mediated cross presentation of antigens to enhance CTL responses. In specific aim 3, we will determine if the adjuvant effect of 5% O2 lysates is due to expression of Annexin A2, a novel putative TLR2 ligand we recently found to be upregulated in hypoxia. Collectively, this knowledge will facilitate groundbreaking approaches to enhance the efficacy of immunotherapy for glioma and other non-central nervous system tumors. PUBLIC HEALTH RELEVANCE: Glioma is an aggressive brain tumor that is very difficult to treat. Vaccines have been tested in glioma patients with suboptimal results. We have developed a novel vaccine with increased efficacy in mouse models. In this project we will optimize the efficacy of this vaccine and elucidate the mechanism by which it works with the long-term goal improving the efficacy of immunotherapy for brain tumor patients.

描述（由申请人提供）：恶性脑肿瘤是35岁以下人群癌症相关死亡的主要原因。个性化疫苗形式的免疫疗法已在选定的胶质瘤患者中表现出免疫活性和临床反应，且毒性最小。有许多正在进行的临床试验，利用培养的肿瘤细胞作为疫苗抗原的来源，用于治疗各种肿瘤。然而，关于细胞培养条件如何影响疫苗接种后的免疫应答和最终临床应答知之甚少。所用的一致条件是肿瘤细胞在大气氧（20%O2）中扩增。我们已经确定了组织培养中使用的氧浓度作为肿瘤细胞疫苗免疫原性的主要决定因素。我们的数据导致了一个有吸引力的中心假设，即肿瘤细胞培养物中的氧张力作为一个主免疫开关，决定了疫苗接种诱导的免疫应答的类型和强度。我们已经可重复地表明，在人和小鼠系统中，相对于在20%O2中培养的神经胶质瘤细胞的裂解物，在5%O2中培养的神经胶质瘤细胞的裂解物引发细胞毒性T淋巴细胞（CTL）具有上级效应子功能。这种差异深刻地影响了免疫治疗的疗效，如胶质瘤和乳腺癌小鼠模型中存活率的显著改善所示。我们证明，相对于20%O2裂解物疫苗，5%O2裂解物疫苗的管理引起上级CTL增殖、细胞因子产生、杀肿瘤功能和运输至肿瘤部位。相反，20%的O2裂解物疫苗增强抗体应答。尽管降低了肿瘤反应性抗体应答，但5%O2疫苗需要B细胞来获得治疗功效，揭示了B细胞在CTL引发中的假定作用。此外，我们有证据表明，在5%O2中生长的胶质瘤细胞上调Toll样受体（TLR）2配体，因为在几个系统中需要TLR 2来区分5%和20%O2裂解物的免疫原性。该提案的目标是阐明“氧开关效应”的分子基础，以合理提高肿瘤细胞疫苗的功效。在具体目标1中，我们将确定和优化调节免疫原性的肿瘤细胞内在变化。增加肿瘤细胞中缺氧诱导因子（HIF）的药理学策略将与随后测定免疫原性的细胞培养物中的实际缺氧进行比较。此外，我们将测试HIF 2a是诱导TLR2配体表达的分子开关的假设。在具体目标2中，我们将检验我们的假设，即需要B细胞是因为IgG-肿瘤裂解物复合物触发Fc受体介导的抗原交叉呈递以增强CTL应答。在具体目标3中，我们将确定5%O2裂解物的佐剂效应是否是由于膜联蛋白A2的表达，膜联蛋白A2是我们最近发现在缺氧中上调的一种新的推定TLR2配体。总的来说，这些知识将促进突破性的方法，以提高神经胶质瘤和其他非中枢神经系统肿瘤的免疫治疗的疗效。 公共卫生关系：神经胶质瘤是一种侵袭性的脑肿瘤，很难治疗。疫苗已经在神经胶质瘤患者中进行了测试，效果不佳。我们已经开发出一种新型疫苗，在小鼠模型中具有更高的功效。在本项目中，我们将优化该疫苗的功效，并阐明其与提高脑肿瘤患者免疫治疗功效的长期目标一起工作的机制。