Antibody cytokine fusion proteins against Cryptococcus neoformans

新型隐球菌抗体细胞因子融合蛋白

基本信息

批准号：
8015629
负责人：
DAVID Owen BEENHOUWER
金额：
$ 31.09万
依托单位：
BRENTWOOD BIOMEDICAL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-03-15 至 2013-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8015629
关键词：
Adjuvant Adverse effects Animals Antibodies Antibody Formation Antigen-Antibody Complex Antigens Attenuated Binding Cells Cellular Immunity Central Nervous System Infections Chemotherapy-Oncologic Procedure Chimeric Proteins Chronic Communicable Diseases Complement Activation Complex Cryptococcal Meningitis Cryptococcus neoformans Cryptococcus neoformans infection Dendritic Cells Disease Dose Effector Cell Encapsulated Equilibrium Fc Receptor Genetic Globulins Goals Granulocyte-Macrophage Colony-Stimulating Factor Granulomatous HIV HIV Infections Heating Human IgG3 Immune response Immunity Immunocompromised Host Immunoglobulin G Immunoglobulin Variable Region Immunotherapeutic agent Infection Interleukin-12 Interleukin-2 Knowledge Link Lung Malignant Neoplasms Modeling Mus Organ Transplantation Organism Persons Play Polysaccharides Populations at Risk Prevention Research Personnel Rheumatism Role Site Surface Talents Testing Therapeutic Therapeutic Uses Toxic effect Tuberculosis Vaccination Vaccine Adjuvant Vaccines Yeasts capsule cytokine design efficacy testing experience flexibility glucuronoxylomannan immunogenic improved killings mortality mouse model novel paracrine pathogen practical application prevent prophylactic protective efficacy receptor binding response therapeutic vaccine tumor uptake vaccination strategy

项目摘要

DESCRIPTION (provided by applicant): Treatment with cytokines is moderately effective against mouse and human infection with Cryptococcus neoformans. However, systemic delivery of cytokines can be problematic for several reasons. Cytokines act in a local manner. Furthermore, they are rapidly degraded and side effects often limit their use. To overcome these limitations, we propose to construct and test several antibody-cytokine fusion proteins against C. neoformans infection with the goal of developing novel prophylactic and therapeutic vaccination strategies. For these fusion proteins, we will use an antibody specific for the outer capsule of C. neoformans, targeting cytokines directly to the site(s) of infection, while improving cytokine stability; thereby increasing the effective dose while decreasing systemic toxicities. There are currently no effective vaccines in use against C. neoformans. By linking cytokines to C. neoformans antigens, antibody-cytokine fusion proteins may be effective in generating protective immunity. Therefore, to test the efficacy of antibody-cytokine fusion proteins in preventing and treating C. neoformans infection, we propose: 1. To construct, produce and characterize antibody-cytokine fusion proteins specific for C. neoformans capsule. Antibody-cytokine fusion proteins consisting of the hinge-flexible human IgG3 against cryptococcal capsular polysaccharide genetically fused to either interleukin-2 (IL-2), granulocyte- macrophage colony stimulating factor (GM-CSF) or IL-12 will be produced recombinantly. Fusion proteins will be characterized for binding to C. neoformans as well as their functionality as cytokines and antibodies. 2. To determine the potential of antibody-cytokine fusion proteins to generate protective immunity to C. neoformans. Heat-killed C. neoformans or capsular polysaccharide will be opsonized with antibody- cytokine fusion proteins described above and administered to mice. Alternatively, dendritic cells will be loaded ex vivo with these immune complexes and delivered to mice. Host antibody and cellular immune responses specific for C. neoformans will be analyzed and protective efficacy of vaccination determined in mouse models of pulmonary and disseminated infection. 3. To determine the efficacy of cytokines genetically fused to antibody specific for C. neoformans capsule in cryptococcal infection. Antibody-cytokine fusion proteins will be given to mice with new or established infection to determine whether they can directly protect against disseminated cryptococcal infection. Host immune responses will be defined and correlated with efficacy. Mortality from cryptococcal meningitis is 10-20% indicating the need for better therapies. The population at risk for cryptococcal disease continues to grow due to increased survival of persons infected with HIV, new HIV infections, and the continued expansion of the number of persons with immunocompromise due to immuno- suppressive therapies for organ transplantation and rheumatic diseases as well as chemotherapy for cancer. The proposed studies have relevance beyond cryptococcal infection, since a significant long-term objective is to design antibody-cytokine fusion proteins for use in other types of infections.

描述（由申请人提供）：用细胞因子的治疗对小鼠和人类的新形象体的感染是适度有效的。但是，由于几个原因，全身递送细胞因子可能会出现问题。细胞因子以局部方式起作用。此外，它们迅速降解，副作用通常会限制其使用。为了克服这些局限性，我们建议构建和测试针对Neoformans感染的几种抗体 - 促动物融合蛋白，目的是开发新型的预防性和治疗性疫苗接种策略。对于这些融合蛋白，我们将使用针对Neoformans外囊的特异性抗体，将细胞因子直接靶向感染部位，同时改善细胞因子稳定性。从而增加有效剂量，同时降低全身毒性。目前尚无针对Neoformans使用的有效疫苗。通过将细胞因子与Neoformans抗原联系起来，抗体 - 促成融合蛋白可能有效地产生保护性免疫。因此，为了测试抗体 - 胞能因子融合蛋白在预防和治疗Neoformans感染中的疗效，我们建议：1。构建，生产和表征针对新生孢梭状芽胞杆菌囊的抗体 - 促动物融合蛋白。由铰链芬太加的人IgG3组成的抗体 - 细胞因子融合蛋白针对遗传上的隐孢子虫囊膜多糖，将其融合到白介素-2（IL-2），粒细胞 - 巨噬细胞刺激因子（GM-CSF）或IL-12中会产生重新构造。融合蛋白将以与新生梭菌的结合及其作为细胞因子和抗体的功能来表征。 2。确定抗体 - 促动物融合蛋白的潜力，以产生对新生梭菌的保护性免疫。热杀死的C. Neoformans或囊囊多糖将用上述抗体 - 细胞因子融合蛋白进行调整，并给予小鼠。或者，将树突状细胞用这些免疫复合物在体内加载并递送至小鼠。将分析针对新生梭菌的宿主抗体和细胞免疫反应，并在肺和传播感染的小鼠模型中确定疫苗接种的保护性疗效。 3。确定细胞因子在遗传上融合的抗体特异性抗体在加密秒感染中的抗体。将向具有新的或已建立的感染的小鼠提供抗体 - 淋巴结融合蛋白，以确定它们是否可以直接防止散布的加密秒感染。宿主免疫反应将被定义并与功效相关。隐球菌脑膜炎的死亡率为10-20％，表明需要更好的疗法。由于感染了HIV感染的人的存活率增加以及由于免疫抑制性疗法的人数增加，由于感染了HIV，新的HIV感染的人的存活增加以及由于对器官移植和风湿性疾病的免疫抑制疗法而持续扩大的人口，持续增长的人口持续增长。拟议的研究具有超出隐球菌感染的相关性，因为一个重要的长期目标是设计抗体 - 促动物融合蛋白，用于其他类型的感染。