Oxygen as a master immunologic switch

氧气作为主要免疫开关

• 批准号: 8695091
• 负责人: CHRISTOPHER A PENNELL
• 金额: $ 30.39万
• 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/8695091
• 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.

描述（由申请人提供）：恶性脑肿瘤是35岁以下患者与癌症相关死亡的主要原因。以个性化疫苗的形式进行免疫疗法表明，在最小毒性的精选神经胶质瘤患者中，免疫活性和临床反应。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，用于治疗多种肿瘤。然而，关于细胞培养条件如何影响疫苗接种后的免疫反应和最终临床反应，知之甚少。使用的一致条件是大气氧中肿瘤细胞的扩展（20％O2）。我们已经确定了在组织培养物中使用的氧浓度是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，即肿瘤细胞培养中的氧张力充当主免疫学开关，决定了疫苗接种引起的免疫反应的类型和强度。我们已经可重复地表明，在5％O2素细胞毒性T淋巴细胞（CTL）中培养的神经胶质瘤细胞的裂解液具有相对于在人类和鼠类系统中20％O2中培养的胶质瘤细胞的裂解液具有出色效应的功能。这种差异深刻影响了免疫疗法的疗效，如鼠神经胶质瘤和乳腺癌的鼠模型的生存率的显着改善所表明。我们证明了5％O2裂解物疫苗接种导致CTL增殖，细胞因子阐述，肿瘤功能以及相对于20％O2裂解物疫苗的肿瘤部位。相反，20％O2裂解物疫苗增强了抗体反应。尽管肿瘤反应性抗体反应降低，但5％O2疫苗需要B细胞才能获得治疗功效，从而揭示了B细胞在CTL启动中的假定作用。此外，我们有证据表明，在5％O2中生长的胶质瘤细胞上调了Toll样受体（TLR）2种配体，因为在几种系统中需要TLR2以区分5％和20％O2裂解物的免疫原性。该提案的目的是阐明“氧开关效应”的分子基础，以合理提高肿瘤细胞疫苗的疗效。在特定目标1中，我们将确定并优化调节免疫原性的肿瘤细胞固有变化。将肿瘤细胞中缺氧诱导因子（HIF）增加的药理策略将与细胞培养物中的实际缺氧进行比较，这些细胞培养物随后将被测定以进行免疫原性。此外，我们将检验以下假设：HIF2A是诱导TLR2配体表达的分子开关。在特定的目标2中，我们将测试我们的假设，即需要B细胞，因为IgG蛋白裂解液配合物触发FC受体介导的抗原的交叉表现以增强CTL反应。在特定的目标3中，我们将确定5％O2裂解物的辅助作用是否是由于膜联蛋白A2的表达是一种新型推定的TLR2配体，我们最近发现在缺氧中被上调。总体而言，这些知识将促进开创性方法，以增强免疫疗法对神经胶质瘤和其他非中心神经系统肿瘤的疗效。