Naso-oropharyngeal Brucella Infections and Mucosal Immune Protection

鼻口咽布鲁氏菌感染与粘膜免疫保护

基本信息

批准号：
9977090
负责人：
David W Pascual
金额：
$ 37.69万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-17 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9977090
关键词：
Aerosols Affect Animals Antibiotic Therapy Arthritis Attenuated Brucella Brucella abortus Brucella melitensis Brucella suis Brucellosis CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Cervical lymph node group Clinical Consumption Contracts Coupled DNA Dairy Products Data Detection Disease Endocarditis FDA approved GTP-Binding Protein alpha Subunits, Gs Goals Human Immune Immune system Immunity Immunization Incidence Induced Abortion Infection Ingestion Interferon Type II Joints Learning Life Link Liver Livestock Lung infections Lymph Node of Head, Face and Neck Lymphoid Tissue Memory Methods Milk Morbidity - disease rate Mucous Membrane Neurologic Nose Oral Oral cavity Oral mucous membrane structure Oropharyngeal Outcome Pathogenesis Pharyngitis Primary Infection Process Quality of life Regimen Resolution Role Route Rural Site Socioeconomic Status Spleen Symptoms Systemic disease Systemic infection T cell response T-Lymphocyte T-Lymphocyte Subsets TNF gene Testing Text Tissues Vaccination Vaccines Virulent Work Zoonoses cell type design draining lymph node flu gastrointestinal symptom human disease improved mutant neglect novel oral infection pathogenic bacteria persistent symptom prevent response socioeconomics tool unpasteurized

项目摘要

ABSTRACT Human brucellosis ranks third among 8 neglected zoonotic diseases, and is contracted as a result of ingesting unpasteurized dairy products. The Gram-negative Brucella is highly infectious, and is believed that less than 2000 CFUs are needed for pulmonary infection. Infection primarily occurs following a mucosal exposure causing a systemic disease manifested by its flu-like symptoms. Despite aggressive antibiotic treatment, it can still persist in a recurring sequelae evident as undulant fever and arthritis. Brucellae survival within the host is linked to its ability to evade intracellular recognition, thus, allowing them to sequester in various tissues. Although human disease is mostly acquired via a mucosal exposure, few studies have examined mucosal immunization for brucellosis. In part, this is attributed to the inefficiency of oral gavage methods requiring an enormous amount of brucellae to induce an infection. Bearing this, we have devised an oral infection method, termed ad bibitum, that retains the infection to the oral mucosa and draining head and neck lymph nodes (HNLNs) recapitulating natural human infections. Hence, we hypothesize that mucosal immunization of the naso-oropharyngeal tissues will derive the required correlates of protective immunity using our novel attenuated Brucella melitensis (BM) strain. Such approaches will allow for the comparison of which immune cell types are needed for protection and to reverse wild-type (wt) BM infection from mounting regulatory responses in order to evade detection. Subsequent to immunization, the attenuated BM mutant stimulates increases in IFN-g- and TNF-a-producing CD4+ and CD8+ T cells, which can effectively eliminate the brucellae. To further these studies, three Specific Aims are proposed. Studies in Specific Aim 1 will establish a mucosal naso-oropharyngeal vaccination regimen that confers protection against mucosal challenge with virulent Brucella. Studies in Specific Aim 2 will determine which T cell subsets are responsible for protection in the mucosal and systemic compartments. Studies in Specific Aim 3 will establish the role for mucosal memory CD8+ T cells in augmenting protection against virulent Brucella challenge more effectively than CD4+ T cells. These studies will further our understanding of how to induce immunity in the naso- oropharyngeal mucosa, and will aid in devising strategies to circumvent BM' evasion methods. This work will ultimately define what constitutes protection and which T cells are needed to guard against wt BM challenges.

抽象的人类布鲁氏菌病在 8 种被忽视的人畜共患疾病中排名第三，因摄入而感染未经高温消毒的乳制品。革兰氏阴性布鲁氏菌具有高度传染性，据信不到 2000 肺部感染需要 CFU。感染主要发生在粘膜暴露后，导致全身性疾病表现为流感样症状。尽管进行了积极的抗生素治疗，但它仍然可能持续存在反复出现明显的后遗症，如波状热和关节炎。布鲁氏菌在宿主体内的存活与其逃避细胞内识别的能力，从而使它们能够隔离在各种组织中。虽然是人类疾病主要是通过粘膜暴露获得的，很少有研究检查粘膜免疫的作用布鲁氏菌病。部分原因是口服灌胃方法效率低下，需要大量的布鲁氏菌以诱发感染。为此，我们设计了一种口腔感染方法，称为 ad bibitum，保留口腔粘膜和引流头颈淋巴结（HNLN）的感染人类自然感染。因此，我们假设鼻口咽组织的粘膜免疫将使用我们的新型减毒布鲁氏菌 (BM) 菌株得出保护性免疫所需的相关性。此类方法将允许比较保护所需的免疫细胞类型并确定逆转野生型 (wt) BM 感染，使其免受监管反应的影响，从而逃避检测。随后的对于免疫接种，减毒 BM 突变体会刺激产生 IFN-g 和 TNF-a 的 CD4+ 和 CD8+ 的增加 T细胞，可有效消灭布鲁氏菌。为了进一步开展这些研究，提出了三个具体目标。具体目标 1 的研究将建立一种可提供保护的粘膜鼻口咽疫苗接种方案对抗有毒布鲁氏菌的粘膜攻击。具体目标 2 中的研究将确定哪些 T 细胞亚群负责粘膜和全身区室的保护。具体目标 3 的研究将建立粘膜记忆 CD8+ T 细胞在更有效地增强针对有毒布鲁氏菌挑战的保护中的作用高于 CD4+ T 细胞。这些研究将进一步加深我们对如何在鼻腔中诱导免疫力的理解。口咽粘膜，并将有助于制定策略来规避 BM 的逃避方法。这项工作将最终定义什么构成保护以及需要哪些 T 细胞来防御 wt BM 挑战。