Immunologic Mechanisms In Experimental Autoimmune Diseases Of The Nervous System

神经系统实验性自身免疫性疾病的免疫机制

• 批准号: 10708596
• 负责人: Steven Jacobson
• 金额: $ 42.58万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10708596
• 关键词: Alzheimer' s Disease; Alzheimer' s disease pathology; Alzheimer' s disease therapeutic; Alzheimer’s disease biomarker; Amyloid beta-Protein; Anatomy; Animal Model; Animals; Antibody Formation; Antibody Response; Antigens; Autoimmune; Autoimmune Diseases of the Nervous System; Autoimmunity; Automobile Driving; Binding; Biological Assay; Biological Markers; Biological Specimen Banks; Biology; Blood Banks; Blood Circulation; Brain; CD8-Positive T-Lymphocytes; Callithrix; Callithrix jacchus jacchus; Central Nervous System; Central Nervous System Diseases; Chronic; Clinical; Clinical Trials; Clinical/Radiologic; Control Animal; Data; Demyelinations; Development; Disease; Disease Progression; Early Diagnosis; Encephalitis; Epilepsy; Exhibits; Experimental Autoimmune Encephalomyelitis; Experimental Models; Exposure to; Future; Generations; Genetic; HHV-6A; HHV-6B; Herpesviridae; Herpesviridae Infections; Human; Human Herpesvirus 6; Immune; Immunize; Immunologics; Infection; Inflammatory; Intravenous; Investigation; Lesion; Liquid substance; Lymphocryptovirus; Lymphoma; Lymphoproliferative Disorders; Magnetic Resonance Imaging; Malignant neoplasm of brain; Mediating; Methods; Microbe; Modality; Modeling; Monitor; Mucous Membrane; Mucous body substance; Multiple Sclerosis; Myelin; National Institute of Neurological Disorders and Stroke; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Nose; Onset of illness; PET/CT scan; Pathogenesis; Pathogenicity; Pathologic; Pathology; Peptides; Peripheral; Peripheral Blood Mononuclear Cell; Phenotype; Physiological; Plasma; Prevalence; Preventive measure; Proteins; Radiology Specialty; Relapse; Reporting; Rodent; Role; Route; Saliva; Senile Plaques; Symptoms; System; T-Lymphocyte Subsets; Testing; Therapeutic; Time; Viral; Viral Antigens; Viral Load result; Viral reservoir; Virus; Virus Diseases; Virus Receptors; X-Ray Computed Tomography; aged; autoreactivity; brain tissue; chemokine; chronic infection; comparative; cytokine; demographics; digital; early onset; experimental analysis; exposed human population; exposure route; gammaherpesvirus; immunogenic; interest; multiple sclerosis treatment; nervous system disorder; neuroinflammation; nonhuman primate; novel therapeutics; pre-clinical; tau Proteins; viral DNA; white matter

We have examined the role of human herpesviruses in CNS disease. In particular, we have focused on the ubiquitous virus HHV-6 (and HHV6B) that is associated with a variety of neurologic diseases including multiple sclerosis, encephalitis, epilepsy, and brain cancer. The generation of an animal model of HHV-6 infection would allow studies on the potential of this virus to cause neurologic disease. Human exposure to HHV-6 occurs most likely through mucous membranes, and we have shown that nasal mucous is a reservoir for the virus. Rodents lack the appropriate receptor for virus binding; however, marmosets are susceptible to HHV6 infection. We have successfully infected marmosets intranasally with HHV-6A, to examine a more physiologic route of exposure with the virus that was comparatively more immunogenic and neuropathogenic. In contrast to what we observed with the intravenous inoculations, marmosets inoculated intranasally with HHV-6A did not exhibit clinical symptoms, did not mount robust anti-HHV-6 antibody responses, and had a marked increase of detectable virus in the periphery (plasma, PBMC, saliva). These data suggested an inverse correlation between antibody production and circulation of viral DNA in the periphery, and moreover, that the clinical symptoms observed in the intravenously inoculated marmosets may have been tied to the robust antibody responses. With this model, we have generated a system in which to independently study the biology of the two HHV-6 species. This is an ongoing problem in the field, due to the high homology between HHV-6A and HHV-6B, the early exposure time to HHV-6B, and the unknown time of exposure to HHV-6A. This also enables us to compare the development of neurologic disease with experimental autoimmune encephalomyelitis (EAE), a well-known established model for MS. EAE is one model of autoimmune inflammatory-mediated CNS demyelination, created by immunizing animals with CNS (usually white matter) peptides or proteins. Although EAE is typically studied in rodents, marmoset EAE presents with greater radiologic and pathologic similarities to MS, reflecting the genetic, immunological, and CNS anatomical proximity of marmosets to humans. EAE in the common marmoset develops cortical and white matter lesions with remarkable immunological and pathological similarity to those seen in MS. We have successfully established EAE in the marmoset, using human white matter homogenate as the antigen(s) to drive inflammatory CNS demyelination, with progressive or relapsing phenotypes. Using these methods of induction, we are able to implement clinical and MRI parameters that will enable us to test new disease modifying therapies. We have demonstrated that intranasal inoculations with HHV-6A and HHV-6B accelerate the MS-like neuroinflammatory disease, EAE. Although those animals inoculated intranasally with HHV-6 were asymptomatic, they exhibited significantly accelerated clinical EAE compared with control animals. In addition, expansion of a proinflammatory CD8+ T cell subsets correlated with post-EAE survival in virus/EAE marmosets, suggesting that a peripheral (viral?) antigen-driven expansion may have occurred post-EAE induction. HHV-6 viral antigen in virus/EAE marmosets was markedly elevated and concentrated in brain lesions, similar to previously reported localizations of HHV-6 in MS brain lesions. Collectively, we demonstrate that asymptomatic intranasal viral acquisition accelerates subsequent neuroinflammation in a nonhuman primate model of MS. Recently, interest in the infectious hypothesis of AD, which suggests that microbes such as herpesviruses may contribute to AD onset or progression, has grown. Novel therapeutics for AD that are successful in rodent preclinical trails often fail in human clinical trials, suggesting that animal models that more closely mimic human AD are needed. Hallmarks of AD pathology, including plaques and tangles, have previously been described as spontaneous pathology in aged marmosets. We are exploring the role of viral infections on AD-related biomarkers in the marmoset model, including pathologic hallmarks, fluid neurodegenerative protein levels, and MRI and PET/CT imaging features. In addition to being competent to experimental HHV-6 infection, marmosets can be naturally infected with CalHV3, an EBV-related gammaherpesvirus and lymphocryptovirus. Similar to human EBV infection, CalHV3 has been associated with chronic persistent infection and, in rare cases, has been associated with lymphoma and lymphoproliferative disease in the marmoset. We have developed a droplet digital PCR (ddPCR) assay that allows for the quantification of CalHV3 viral load in infected animals. We have characterized prevalence and demographics associated with CalHV3 infection in the NINDS colony in order to further explore the utility of CalHV3 infection as a model of chronic herpesvirus infection and its potential role in neurodegenerative disease. Using the marmoset model of experimental HHV-6 infection and natural CalHV3 infection, we are investigating the effects of herpesvirus infection on biomarkers of AD in a translationally relevant comparative animal model. We are identifying and quantifying levels of AD-related hallmarks, such as beta-amyloid and tau, in our extensive marmoset tissue brain and blood bank specimens, including both herpesvirus infected and uninfected marmosets to determine if viral infection enhances biomarker levels, which would suggest a role for herpesviruses in AD pathogenesis and help dictate future preventative measures and disease therapeutics.

我们已经检查了人类疱疹病毒在中枢神经系统疾病中的作用。特别是，我们专注于无处不在的病毒HHV-6（和HHV6B），该病毒与多种神经系统疾病有关，包括多发性硬化症，脑炎，癫痫和脑癌。 HHV-6感染的动物模型的产生将允许研究该病毒引起神经系统疾病的潜力。人类对HHV-6的暴露最有可能通过粘膜发生，我们已经表明鼻粘液是该病毒的储层。啮齿动物缺乏适当的病毒结合受体。但是，摩尔果会容易受到HHV6感染的影响。我们已经成功地用HHV-6A在鼻内感染了Marmosets，以检查与病毒相对较高的免疫原性和神经病的病毒的更生理的暴露途径。与我们从静脉接种的观察到的相反，与HHV-6A室内接种的摩尔糖不会表现出临床症状，没有固定强大的抗HHV-6抗体反应，并且在外围（Plasma，PBMC，saliva）中有明显的可检测病毒增加。这些数据表明，抗体产生与外围病毒DNA的循环之间存在反相关性，此外，在静脉接种的果胶中观察到的临床症状可能与稳健的抗体反应有关。使用该模型，我们生成了一个系统，可以独立研究这两种HHV-6物种的生物学。由于HHV-6A和HHV-6B之间的同源性很高，早期暴露于HHV-6B以及未知的HHV-6A暴露时间，这是该领域的持续问题。这也使我们能够将神经系统疾病的发展与实验性自身免疫性脑脊髓炎（EAE）进行比较，这是MS的众所周知的既定模型。 EAE是一种自身免疫性炎症介导的中枢神经系统的模型，它是通过用中枢神经系统（通常是白质）肽或蛋白质免疫动物而创建的。尽管通常在啮齿动物中研究了EAE，但Marmoset EAE与MS具有更大的放射学和病理相似性，反映了Marmosets与人类的遗传，免疫学和中枢神经系统的解剖结构。 公共摩尔果中的EAE与MS中发现的皮质和白质病变具有显着的免疫学和病理相似性。我们已经成功地建立了在马莫斯特（Marmoset），使用人类白质匀浆作为抗原，以促进炎症性中枢神经系统脱髓鞘，并具有渐进式或复发的表型。使用这些诱导方法，我们能够实施临床和MRI参数，从而使我们能够测试新的疾病修饰疗法。我们已经证明，HHV-6A和HHV-6B的鼻内接种加速了MS样神经炎性疾病，EAE。尽管这些动物与HHV-6内鼻内接种了无症状，但与对照动物相比，它们表现出显着加速的临床EAE。此外，促炎的CD8+ T细胞子集的扩展与病毒/EAE果marmoset中的-ee后存活相关，这表明可能发生了外周（病毒？）抗原驱动的扩张。病毒/EAE Marmoset中的HHV-6病毒抗原显着升高并集中在脑损伤中，类似于先前报道的HHV-6在MS脑病变中的局部局部定位。总体而言，我们证明了无症状的鼻内病毒采集会在MS的非人类灵长类动物模型中加速随后的神经炎症。 最近，对AD的感染假设的兴趣表明，诸如疱疹病毒之类的微生物可能有助于AD发作或进展。在啮齿动物的临床前踪迹中成功的AD的新型AD治疗方法在人类的临床试验中常常失败，这表明需要更紧密地模拟人类AD的动物模型。 AD病理学的标志，包括斑块和缠结，以前被描述为老年棉花糖的自发病理。我们正在探索病毒感染在Marmoset模型中与AD相关生物标志物的作用，包括病理标志，流体神经退行性蛋白水平以及MRI和PET/CT成像特征。除了具有实验性HHV-6感染能力之外，Marmoset还可以自然地感染EBV相关的伽马鞘病毒和淋巴结病毒。与人类EBV感染相似，CALHV3与慢性持续感染有关，在极少数情况下，与Marmoset中的淋巴瘤和淋巴细胞增生性疾病有关。我们已经开发了一种液滴数字PCR（DDPCR）测定法，该测定法可以定量受感染动物的CALHV3病毒负荷。我们已经表征了与CALHV3感染相关的患病率和人口统计学，以便进一步探索CalHV3感染作为慢性疱疹病毒感染及其在神经退行性疾病中的潜在作用的模型。使用实验性HHV-6感染和天然CALHV3感染的Marmoset模型，我们正在研究疱疹病毒感染对翻译相关的比较动物模型中AD生物标志物的影响。我们正在我们广泛的果绿色组织脑和血库标本中确定和量化与广告相关的标志水平，例如β-淀粉样蛋白和tau，包括感染和未感染的疱疹病毒，以确定病毒感染能够增强生物标志物的水平，这会提高生物标志性的疾病，并有助于培养疱疹病毒疾病。