Perspectives on the Intracellular Bacterium Chlamydia pneumoniae in Late-Onset Dementia

Purpose of Review Chronic diseases remain a daunting challenge for clinicians and researchers alike. While difficult to completely understand, most chronic diseases, including late-onset dementias, are thought to arise as an interplay between host genetic factors and environmental insults. One of the most diverse and ubiquitous environmental insults centers on infectious agents. Associations of infectious agents with late-onset dementia have taken on heightened importance, including our investigations of infection by the intracellular respiratory bacterium, Chlamydia pneumoniae (Cpn), in late-onset dementia of the Alzheimer’s type. Recent Findings Over the last two decades, the relationship of this infection to pathogenesis in late-onset dementia has become much clearer. This clarity has resulted from applying contemporary molecular genetic, biochemical, immunochemical, and cell culture techniques to analysis of human brains, animal models, and relevant in vitro cell culture systems. Data from these studies, taken in aggregate form, now can be applied to evaluation of proof of concept for causation of this infection with late-onset disease. In this evaluation, modifications to the original Koch postulates can be useful for elucidating causation. Summary All such relevant studies are outlined and summarized in this review, and they demonstrate the utility of applying modified Koch postulates to the etiology of late-onset dementia of the Alzheimer’s type. Regardless, it is clear that even with strong observational evidence, in combination with application of modifications of Koch’s postulates, we will not be able to conclusively state that Cpn infection is causative for disease pathogenesis in late-onset dementia. Moreover, this conclusion obtains as well for the putative causation of this condition by other pathogens, including herpes simplex virus type 1, Borrelia burgdorferi, and Porphyromonas gingivalis

Chlamydia pneumoniae Infection of Monocytes in vitro Stimulates Innate and Adaptive Immune Responses Relevant to those in Alzheimer\u27s Disease.

Background: Alzheimer\u27s disease (AD) is a progressive neurodegenerative disorder in which infection with Chlamydia pneumoniae (Cpn) has been associated. Cpn is an obligate intracellular respiratory pathogen that may enter the central nervous system (CNS) following infection and trafficking of monocytes through the blood-brain barrier. Following this entry, these cells may secrete pro-inflammatory cytokines and chemokines that have been identified in the AD brain, which have been thought to contribute to AD neurodegeneration. The objectives of this work were: (i) to determine if Cpn infection influences monocyte gene transcript expression at 48 hours post-infection and (ii) to analyze whether pro-inflammatory cytokines are produced and secreted from these cells over 24 to 120 hours post-infection. Methods: Gene transcription was analyzed by RT-PCR using an innate and adaptive immunity microarray with 84 genes organized into 5 functional categories: inflammatory response, host defense against bacteria, antibacterial humoral response, septic shock, and cytokines, chemokines and their receptors. Statistical analysis of the results was performed using the Student\u27s t-test. P-values ≤ 0.05 were considered to be significant. ELISA was performed on supernatants from uninfected and Cpn-infected THP1 monocytes followed by statistical analysis with ANOVA. Results: When Cpn-infected THP1 human monocytes were compared to control uninfected monocytes at 48 hours post-infection, 17 genes were found to have a significant 4-fold or greater expression, and no gene expression was found to be down-regulated. Furthermore, cytokine secretion (IL-1ß, IL-6, IL-8) appears to be maintained for an extended period of infection. Conclusions: Utilizing RT-PCR and ELISA techniques, our data demonstrate that Cpn infection of THP1 human monocytes promotes an innate immune response and suggests a potential role in the initiation of inflammation in sporadic/late-onset Alzheimer\u27s disease

Evaluation of Smell and miRNA Biomarkers in Alzheimer’s Disease (AD)

Abstract and poster under embargo until May 31, 2019

Role of Microbes in the Development of Alzheimer\u27s Disease: State of the Art - An International Symposium Presented at the 2017 IAGG Congress in San Francisco.

This article reviews research results and ideas presented at a special symposium at the International Association of Gerontology and Geriatrics (IAGG) Congress held in July 2017 in San Francisco. Five researchers presented their results related to infection and Alzheimer\u27s disease (AD). Prof. Itzhaki presented her work on the role of viruses, specifically HSV-1, in the pathogenesis of AD. She maintains that although it is true that most people harbor HSV-1 infection, either latent or active, nonetheless aspects of herpes infection can play a role in the pathogenesis of AD, based on extensive experimental evidence from AD brains and infected cell cultures. Dr. Miklossy presented research on the high prevalence of bacterial infections that correlate with AD, specifically spirochete infections, which have been known for a century to be a significant cause of dementia (e.g., in syphilis). She demonstrated how spirochetes drive senile plaque formation, which are in fact biofilms. Prof. Balin then described the involvement of brain tissue infection by th

Detection of Bacterial Antigens and Alzheimer\u27s Disease-like Pathology in the Central Nervous System of BALB/c Mice Following Intranasal Infection with a Laboratory Isolate of Chlamydia pneumoniae

Pathology consistent with that observed in Alzheimer’s disease (AD) has previously been documented following intranasal infection of normal wild-type mice with Chlamydia pneumoniae (Cpn) isolated from an AD brain (96-41). In the current study, BALB/c mice were intranasally infected with a laboratory strain of Cpn, AR-39, and brain and olfactory bulbs were obtained at 1-4 months post-infection (pi). Immunohistochemistry for amyloid beta or Cpn antigens was performed on sections from brains of infected or mock-infected mice. Chlamydia-specific immunolabeling was identified in olfactory bulb tissues and in cerebrum of AR-39 infected mice. The Cpn specific labeling was most prominent at 1 month pi and the greatest burden of amyloid deposition was noted at 2 months pi, whereas both decreased at 3 and 4 months. Viable Cpn was recovered from olfactory bulbs of 3 of 3 experimentally infected mice at 1 and 3 months pi, and in 2 of 3 mice at 4 months pi. In contrast, in cortical tissues of infected mice at 1 and 4 months pi no viable organism was obtained. At 3 months pi, only 1 of 3 mice had a measurable burden of viable Cpn from the cortical tissues. Mock-infected mice (0 of 3) had no detectable Cpn in either olfactory bulbs or cortical tissues. These data indicate that the AR-39 isolate of Cpn establishes a limited infection predominantly in the olfactory bulbs of BALB/c mice. Although infection with the laboratory strain of Cpn promotes deposition of amyloid beta, this appears to resolve following reduction of the Cpn antigen burden over time. Our data suggest that infection with the AR-39 laboratory isolate of Cpn results in a different course of amyloid beta deposition and ultimate resolution than that observed following infection with the human AD-brain Cpn isolate, 96-41. These data further support that there may be differences, possibly in virulence factors, between Cpn isolates in the generation of sustainable AD pathology

Effect of Age and Vaccination on Extent and Spread of Chlamydia pneumoniae Infection in C57BL/6 Mice

BACKGROUND: Chlamydia pneumoniae is an obligate intracellular respiratory pathogen for humans. Infection by C. pneumoniae may be linked etiologically to extra-respiratory diseases of aging, especially atherosclerosis. We have previously shown that age promotes C. pneumoniae respiratory infection and extra-respiratory spread in BALB/c mice. FINDINGS: Aged C57BL/6 mice had a greater propensity to develop chronic and/or progressive respiratory infections following experimental intranasal infection by Chlamydia pneumoniae when compared to young counterparts. A heptavalent CTL epitope minigene (CpnCTL7) vaccine conferred equal protection in the lungs of both aged and young mice. This vaccine was partially effective in protecting against C. pneumoniae spread to the cardiovascular system of young mice, but failed to provide cardiovascular protection in aged animals. CONCLUSIONS: Our findings suggest that vaccine strategies that target the generation of a C. pneumoniae-specific CTL response can protect the respiratory system of both young and aged animals, but may not be adequate to prevent dissemination of C. pneumoniae to the cardiovascular system or control replication in those tissues in aged animals

Comparison of Chlamydia antigen and AD-like pathology in the brains of BALB/c mice following intranasal infection with Chlamydia muridarum or Chlamydia pneumoniae

Previous research indicates BALB/c mice inoculated with Chlamydia pneumoniae (Cpn) demonstrated AD-like pathology which suggests that this mouse model is valid for studying the pathogenesis implicated in Alzheimer’s disease (AD). Studies have demonstrated that Chlamydia trachomatis (Ctr) can disseminate from its primary site of infection and plays a major role in the induction of reactive arthritis. The objectives of this lab are: (1) to identify and localize Chlamydia antigens in the brains of BALB/c mice infected with C. muridarum and (2) to determine if infection with C. muridarum induces AD-like pathology comparable to Cpn. Using mouse adapted respiratory isolates of C. muridarum, we investigated whether C. muridarum disseminated from the respiratory tract to the brain. Mice were intranasally infected with plaqued C small Weiss (CSW) or plaqued mouse pneumonitis Weiss (MoPn Weiss). Brain tissue was isolated at 2 months post-infection. Serial sections from brains infected mice were analyzed for amyloid or Chlamydia antigens. Preliminary analysis of brain tissue demonstrated no detectable difference in C. muridarum antigen between mice receiving 1 x105 IFU and mice receiving 1 x101 IFU, whereas a small but detectable difference was identified in amyloid-specific labeling between these two experimental groups. In contrast, prominent Chlamydia-specific labeling was identified in the brains of Cpn-infected mice as well as substantial amyloid deposition at 2 months p.i.. These data suggest that, relative to Cpn AR-39 infection, C. muridarum infection is a weaker stimulus for inflammation, resulting in decreased amyloid deposition in the brains of BALB/c mice

Chlamydia pneumoniae: An Etiologic Agent for Late-Onset Dementia

The disease known as late-onset Alzheimer\u27s disease is a neurodegenerative condition recognized as the single most common form of senile dementia. The condition is sporadic and has been attributed to neuronal damage and loss, both of which have been linked to the accumulation of protein deposits in the brain. Significant progress has been made over the past two decades regarding our overall understanding of the apparently pathogenic entities that arise in the affected brain, both for early-onset disease, which constitutes approximately 5% of all cases, as well as late-onset disease, which constitutes the remainder of cases. Observable neuropathology includes: neurofibrillary tangles, neuropil threads, neuritic senile plaques and often deposits of amyloid around the cerebrovasculature. Although many studies have provided a relatively detailed knowledge of these putatively pathogenic entities, understanding of the events that initiate and support the biological processes generating them and the subsequent observable neuropathology and neurodegeneration remain limited. This is especially true in the case of late-onset disease. Although early-onset Alzheimer\u27s disease has been shown conclusively to have genetic roots, the detailed etiologic initiation of late-onset disease without such genetic origins has remained elusive. Over the last 15 years, current and ongoing work has implicated infection in the etiology and pathogenesis of late-onset dementia. Infectious agents reported to be associated with disease initiation are various, including several viruses and pathogenic bacterial species. We have reported extensively regarding an association between late-onset disease and infection with the intracellular bacterial pathoge

Proof of Concept Studies of Chlamydia Pneumoniae Infection as a Trigger for Late-onset Alzheimer Disease

Aims Objectives: Chronic infection(s) may be initiators for Alzheimer disease pathogenesis. Our laboratory has shown that Chlamydia pneumoniae infects cells in vitro, can survive intracellularly for long periods of time, and is detectable in Alzheimer disease brain tissues. Proof of concept studies are being performed to determine how infection may enter the brain, influence protein and gene regulation, and activate neuroinflammation observed in Alzheimer disease. Method Animal (BALB/c mice) and cellular studies are evaluating entry of Chlamydia pneumoniae into the brain and human monocytic cells, respectively, and the consequences of infection with regards to gene and protein expression. Proteins and genes involved with AD pathology and neuroinflammation are being analyzed using commercial human ELISA and qRTPCR Array approaches. Results Amyloid turn-on (Abeta 1-42) occurs in both animal brains and human cells following Chlamydia pneumoniae infection. Genes and proteins corresponding to neuroinflammatory markers and inflammasomes from infectied cells (eg, THP1 monocytes) are up-regulated when compared with uninfected monocytes. Of those genes and proteins up-regulated, CCL2, IL-1beta, AIM2, and IDO are significantly changed and reflect the activated pro-inflammatory state of infected monocytes. Conclusion Our data suggest that Chlamydia pneumoniae introduced intranasally to normal BALB/c mice triggers the accumulation of Abeta 1-42 in the brain. Further, upon infection of THP-1 monocytes, significant changes occur in the regulation of gene transcripts and protein expression that correlates to the inflammatory signature observed in late-onset Alzheimer disease. Thus, our data suggest that infection can trigger specific protein and gene changes that correlate to Alzheimer disease pathogenesis