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.

摘要 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。 未经巴氏灭菌的乳制品。革兰氏阴性布鲁氏菌具有很高的传染性，据信不到2000 肺部感染需要CFU。感染主要发生在黏膜暴露后，导致 表现为流感样症状的全身性疾病。尽管进行了积极的抗生素治疗，但它仍然可以持续 在反复发作的后遗症中，明显表现为波浪性发热和关节炎。布鲁氏菌在宿主体内的存活与其 能够逃避细胞内的识别，从而使它们能够在各种组织中隔离。虽然是人类 疾病主要是通过粘膜接触而获得的，很少有研究研究粘膜免疫对 布鲁氏菌病。这在一定程度上归因于口服灌胃方法效率低下，需要大量的 导致感染的布鲁氏菌。为此，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和引流头颈部淋巴结(HNLN)的重述 自然的人类感染。因此，我们假设鼻咽部组织的粘膜免疫 将使用我们的新型减毒羊种布鲁氏菌(BM)菌株推导出所需的保护性免疫相关系数。 这种方法将允许比较需要哪些免疫细胞类型来保护和 逆转野生型(Wt)BM感染增加的调控反应，以逃避检测。后续 在免疫过程中，减毒的BM突变体刺激产生干扰素-g-和肿瘤坏死因子-α的CD4+和CD8+增加 T细胞，可有效消除布鲁氏菌。为了进一步开展这些研究，本文提出了三个具体目标。 针对特定目标1的研究将建立一种提供保护的鼻咽部黏膜疫苗接种方案 抵抗布氏杆菌毒力的黏膜攻击。对特定目标2的研究将确定哪些T细胞亚群是 负责粘膜和全身隔间的保护。对具体目标3的研究将建立 粘膜记忆CD8+T细胞在更有效地增强对布鲁氏菌毒力攻击的保护作用 而不是CD4+T细胞。这些研究将进一步加深我们对如何在鼻腔内诱导免疫的理解。 口咽粘膜，并将有助于制定策略，以规避BM的逃避方法。这项工作将 最终确定什么是保护，哪些T细胞是预防wt BM挑战所必需的。