Antigen discovery and validation in Pneumocystis pneumonia

肺孢子虫肺炎抗原的发现和验证

• 批准号: 8782716
• 负责人: Taylor John Eddens
• 金额: $ 4.77万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8782716
• 关键词: AIDS/HIV problem; Adoptive Transfer; Antibodies; Antibody Formation; Antigens; Ants; Area; Autoimmune Process; Autoimmunity; B-Lymphocytes; Biological Assay; Biotinylation; C57BL/6 Mouse; CD19 gene; Clinical; Complement; Consensus; Cyst; DNA; DNA Vaccines; Data; Development; Diagnosis; Diagnostic tests; Diffuse; Disease; Emerging Communicable Diseases; Enzymes; Exanthema; Extracellular Protein; Flow Cytometry; Fungal Proteins; Future; Gene Expression Profile; Glomerulonephritis; Goals; HIV; Hematologic Neoplasms; Histones; Immune response; Immunocompromised Host; Immunoglobulin G; Immunologic Deficiency Syndromes; Immunologics; Immunosorbents; Immunosuppressive Agents; In Vitro; Individual; Infection; Integral Membrane Protein; Interstitial Pneumonia; Lead; Life; Life Cycle Stages; Link; Lung; Mediastinal lymph node group; Mediating; Memory; Memory B-Lymphocyte; Mission; Monoclonal Antibodies; Morbidity - disease rate; Mus; National Institute of Allergy and Infectious Disease; Nature; Opportunistic Infections; Patients; Peptides; Pharmaceutical Preparations; Pharmacotherapy; Pneumocystis; Pneumocystis carinii Pneumonia; Pneumonia; Populations at Risk; Proteins; Proteome; Proteomics; RNA Sequences; Regimen; Reproduction spores; Serum; Shapes; Spottings; Surface; T-Lymphocyte; Therapeutic immunosuppression; Training; Transplantation; Vaccinated; Vaccination; Vaccines; Validation; base; design; expression vector; extracellular; fungus; immunogenicity; killings; macrophage; mortality; novel; novel strategies; public health relevance; research study; response; therapeutic development; therapy development; vaccine development

DESCRIPTION (provided by applicant): Pneumocystis (PC) is a dimorphic fungus that causes pneumonia in immunocompromised individuals and PC pneumonia (PCP) is the most common serious opportunistic infection in individuals diagnosed with HIV. Furthermore, PCP is re-emerging in the clinical setting of non-HIV infected immunocompromised individuals receiving life-saving immunosuppressive therapies for conditions such as hematological malignancies, transplantation, and autoimmune conditions. As the at-risk population continues to increase, finding novel, conserved, extracellular antigens on PC is the first step towards designing more effective diagnostic tests, creating monoclonal antibody based therapies, and developing vaccines. To discover novel antigens, proteomic and transcriptome analysis of PC was conducted. Specifically, surface biotinylation of total PC, containing both the cyst ant troph life form, was performed and demonstrated that histone 2B (H2B) was an abundant extracellular protein on PC. RNA sequencing was also performed on separated PC cysts and trophs to identify targets differentially expressed in the two life forms. H2B had a 3-fold increase in expression in the cyst form when compared to the troph form, while other proteins, such as a Meu10, were found to have higher expression in the troph form. To determine the immunogenicity of H2B, mice were vaccinated with an H2B DNA vaccine. Mice receiving the H2B vaccine had a significant increase in anti-PC serum IgG and had a significant decrease in lung PC burden when compared to unvaccinated controls. These experiments demonstrate that H2B is a cyst-enriched antigen and is capable of providing protection against developing PCP following DNA vaccination. The aims of this proposed project are: 1.) To define the humoral immune response generated by H2B DNA vaccination, and 2.) To determine the immunogenicity of a Meu10 DNA vaccine and evaluate the protection provided by a life cycle based multivalent DNA vaccination. To accomplish these aims, the proposed studies will explore the memory B cell response generated by H2B DNA vaccination and examine the immunologic mechanisms of protection provided by the vaccine. Furthermore, vaccination studies will be conducted with a Meu10 DNA vaccine and a multivalent vaccine targeting both the cyst-enriched H2B and the troph-enriched Meu10 to examine the efficacy of a targeted, life cycle based multivalent vaccine. The proposed studies will define the fundamental immunological mechanisms behind a previously designed H2B vaccine and will explore a novel strategy for vaccine development for the re- emerging infectious disease of PC in accordance with the mission statement of the NIAID.

描述（由申请人提供）：肺炎（PC）是一种二态真菌，可在免疫功能低下的个体和PC肺炎（PCP）中引起肺炎（PCP）是诊断为HIV的个体中最常见的严重机会感染。此外，在非HIV感染的免疫功能低下的个体的临床环境中，PCP正在重新出现，以接受挽救生命的免疫抑制疗法，例如血液学恶性肿瘤，移植和自身免疫性条件。随着高危人群的继续增加，在PC上找到新颖，保守的细胞外抗原是设计更有效的诊断测试，创建基于单克隆抗体的疗法并开发疫苗的第一步。为了发现新型抗原，对PC进行了蛋白质组学和转录组分析。具体而言，总PC的表面生物素化，既包含囊肿蚂蚁滋养寿命 进行形式，并证明组蛋白2b（H2b）是PC上的丰富细胞外蛋白。还对分离的PC囊肿和TROPH进行了RNA测序，以识别以两种生命形式差异表达的靶标。与Troph形式相比，H2B的囊肿形式表达增加了3倍，而其他蛋白质（例如MEU10）在Troph形式中的表达较高。为了确定H2B的免疫原性，用H2B DNA疫苗接种小鼠。接受H2B疫苗的小鼠与未接种疫苗的对照相比，抗PC血清IgG的抗PC血清IgG显着增加，肺PC负担显着减少。这些实验表明，H2B是一种富含囊肿的抗原，能够在DNA疫苗接种后提供防止开发PCP的保护。该提出的项目的目的是：1。）定义由H2B DNA疫苗接种产生的体液免疫反应，以及2.）确定MEU10 DNA疫苗的免疫原性，并评估由基于生命周期的多价DNA疫苗提供的保护。为了实现这些目的，拟议的研究将探索由H2B DNA疫苗接种产生的记忆B细胞反应，并检查疫苗提供的保护机理。此外，将使用MEU10 DNA疫苗和靶向富含囊肿的H2B的多价疫苗进行疫苗接种研究，以及富含Troph的MEU10检查目标生命周期基于多价疫苗的功效。拟议的研究将定义先前设计的H2B疫苗背后的基本免疫机制，并将根据NIAID的使命陈述，探索一种新型PC传染病的疫苗开发策略。