Naso-oropharyngeal Brucella Infections and Mucosal Immune Protection

鼻口咽布鲁氏菌感染与粘膜免疫保护

• 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
• 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

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 人布鲁氏菌病在8种被忽视的人畜共患病中排名第三，由于摄入 未经巴氏消毒的乳制品。革兰氏阴性布鲁氏菌具有很高的感染力，被认为不到2000年 肺部感染需要CFU。感染主要发生在导致粘膜暴露后发生 全身性疾病以其流感样症状表现出来。尽管有侵略性的抗生素治疗，但它仍然可以持续 在反复出现的后遗症中，显然是起伏的热和关节炎。宿主内的布鲁氏菌生存与其 因此，能够逃避细胞内识别，从而使它们可以隔离在各种组织中。虽然人类 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 布鲁氏菌病。这部分归因于口服方法的效率低下，需要大量数量 布鲁氏菌会诱发感染。考虑到这一点，我们设计了一种口腔感染方法，称为ad Bibitum， 保留对口腔粘膜的感染以及排水头和颈部淋巴结（HNLNS） 天然人类感染。因此，我们假设鼻孔 - 咽组织的粘膜免疫 将使用我们新型的衰减布鲁氏菌（BM）菌株来得出保护性免疫所需的相关性。 这种方法将允许比较哪种免疫细胞类型来保护和 逆转野生型（WT）BM感染从安装调节反应中以逃避检测。随后的 为了进行免疫，减毒的BM突变体刺激IFN-G-和TNF-A产生CD4+和CD8+的增加 T细胞，可以有效地消除布鲁氏菌。为了进一步研究这些研究，提出了三个具体目标。 特定目标1中的研究将建立一种赋予保护的粘膜鼻咽咽疫苗疫苗接种方案 反对有毒布鲁氏菌的粘膜挑战。特定目标2中的研究将确定哪些T细胞子集为 负责在粘膜和系统隔室中进行保护。特定目标3的研究将确定 粘膜记忆CD8+ T细胞的作用在增强对毒性布鲁氏菌挑战的保护中的作用更有效 比CD4+ T细胞。这些研究将进一步了解如何在鼻腔中诱导免疫力 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 最终定义构成保护的方法以及需要哪些T细胞来防止WT BM挑战。