Chlamydophila (Chlamydia) pneumoniae promotes Ab 1-42 amyloid processing in Neuronal Cells: A Pathogenic Trigger for Alzheimer\u27s Disease

Background: Previously, our laboratory identified Chlamydophila (Chlamydia) pneumoniae (Cpn) in autopsied sporadic AD brains. Furthermore, we have developed a BALB/c mouse model that demonstrated infection-induced amyloid plaques similar to those found in AD, and demonstrated that Cpn infection of neuronal cells inhibited apoptotic pathways of cell death. Hypothesis: Our current studies address whether infection with Cpn in neuronal cells triggers abnormal cleavage of the beta amyloid precursor protein (bAPP) into Ab1-42, thereby contributing to amyloid plaque formation characteristic of the pathology identified in AD. Materials and Methods: Human neuroblastoma cells were infected with the respiratory strain AR39 Cpn in vitro, then amyloid processing was analyzed and quantitated using immunocytochemistry, Western blotting and ELISA assays. Results: Cpn was shown to infect neuronal cells and induce intracellular amyloid processing. Cpn infection yielded cytoplasmic labeling of Ab 1-42 that was increased relative to uninfected cells. The ELISA assay revealed that in neuronal cell lysates, Ab 1-42 in the infected cells was increased 3 to 16-fold over the uninfected cells, from 24 to 72hr post infection. Western blot analysis confirmed an increase in Ab 1-42 in the infected neuronal cell lysates. Conclusions: These data suggest that infection of neuronal cells with Chlamydophila (Chlamydia) pneumoniae alters the processing of bAPP, thereby producing Ab1-42. Therefore, these studies and previous research reported by our laboratory support the implication of Cpn as a pathogenic agent in perpetuating the hallmark amyloid plaque formations observed in AD. This concept holds major therapeutic considerations for future studies.https://digitalcommons.pcom.edu/posters/1004/thumbnail.jp

Reducing the stigma of mental illness among medical students

Background: The American Osteopathic Association House of Delegates Resolution 205 recommends “increased awareness of depression amongst U.S. medical students” due to the increasing body of research describing the rise of depression, burn-out and suicide ideation among medical students. There is consequently a need to understand mental health issues as a component of professional development. Hypothesis: A student-led symposium addressing mental and emotional health topics relevant to medical students will reduce the stigma associated with mental illness. Materials and Methods: A 2-hour student-run “Patient Perspective” session was held during the second year neuroscience block in the PCOM DO program. One week before the program, a student-developed, online Wellness Survey measured prevalence of mental illness, common feelings during medical school, coping mechanisms used for stress, and use of mental health resources. Immediately before and after the program, students were asked to report their familiarity with mental illness and their feelings regarding a vignette about a mentally ill woman. Pre- and post-activity surveys were provided by the University of California San Francisco School of Medicine and adapted for the event. During the program, data from the online survey were shared, student organizers discussed emotional wellness and positive coping mechanisms in the context of the profession, and student panelists shared their experiences with mental health issues. A faculty psychiatrist spoke about mental health resources, and attendees received pamphlets listing these resources. The event concluded with student-led breakout sessions in which stress during medical school and strategies for promoting positive coping mechanisms were discussed, followed by administration of the post-activity survey. Results: 113 students completed the pre-activity survey; 89 completed the post-activity survey. For these 89, differences between responses were universally in the direction of increasing acceptance and decreasing stigma of those with mental illness; all differences were statistically significant. The largest shift regarded students’ reluctance to disclose their own theoretical mental illness to colleagues. Conclusion: Incorporating an emotional health symposium into medical students’ training may increase understanding and acceptance of those who may have mental illness and reduce stigma associated with mental illness

Infection with Chlamydia Pneumoniae Alters Calcium-associated Gene Regulation and Processes in Neuronal Cells and Monocytes: Implications for Alzheimer’s Disease

Background: First proposed by Khachaturian in 1994, the calcium hypothesis postulates that sustained disturbance of intracellular calcium is the leading cause of neurodegenerative disorders. Studies showing alteration in calcium signaling in both sporadic and familial Alzheimer’s disease (AD) support this hypothesis. Intracellular calcium signaling is tightly regulated in time, intensity, and space, and is responsible for a variety of neuronal functions. Calcium influx from the extracellular environment modulates calcium levels, as do intracellular stores in the endoplasmic reticulum. The focus of this study was to test various calcium related genes in both monocytes and neuronal cells. Previous studies have shown that cells infected with Chlamydia pneumoniae (Cpn) exhibit altered protein processing, such as amyloid and tau modification, consistent with those found in AD. We expect to see significant alterations in calcium genes, as well as their protein products in Cpn infected cells. Every calcium gene has a unique function in the cell. Determining which genes are up or down regulated following infection may provide insight into how the neurodegeneration process observed in AD is initiated by Cpn infections

Immunohistological detection of Chlamydia pneumoniae in the Alzheimer's disease brain

<p>Abstract</p> <p>Background</p> <p>Sporadic late-onset Alzheimer's disease (AD) appears to evolve from an interplay between genetic and environmental factors. One environmental factor that continues to be of great interest is that of <it>Chlamydia pneumoniae </it>infection and its association with late-onset disease. Detection of this organism in clinical and autopsy samples has proved challenging using a variety of molecular and histological techniques. Our current investigation utilized immunohistochemistry with a battery of commercially available anti-<it>C. pneumoniae </it>antibodies to determine whether <it>C. pneumoniae </it>was present in areas typically associated with AD neuropathology from 5 AD and 5 non-AD control brains.</p> <p>Results</p> <p>Immunoreactivity for <it>C. pneumoniae </it>antigens was observed both intracellularly in neurons, neuroglia, endothelial cells, and peri-endothelial cells, and extracellularly in the frontal and temporal cortices of the AD brain with multiple <it>C. pneumoniae</it>-specific antibodies. This immunoreactivity was seen in regions of amyloid deposition as revealed by immunolabeling with two different anti-beta amyloid antibodies. Thioflavin S staining, overlaid with <it>C. pneumoniae </it>immunolabeling, demonstrated no direct co-localization of the organism and amyloid plaques. Further, the specificity of <it>C. pneumoniae </it>labeling of AD brain sections was demonstrated using <it>C. pneumoniae </it>antibodies pre-absorbed against amyloid β 1-40 and 1-42 peptides.</p> <p>Conclusions</p> <p>Anti-<it>C. pneumoniae </it>antibodies, obtained commercially, identified both typical intracellular and atypical extracellular <it>C. pneumoniae </it>antigens in frontal and temporal cortices of the AD brain. <it>C. pneumoniae</it>, amyloid deposits, and neurofibrillary tangles were present in the same regions of the brain in apposition to one another. Although additional studies are required to conclusively characterize the nature of Chlamydial immunoreactivity in the AD brain, these results further implicate <it>C. pneumoniae </it>infection with the pathogenesis of Alzheimer's disease.</p

Reducing the Stigma of Mental Illness Among Medical Students

Abstract: The American Osteopathic Association House of Delegates Resolution 205 recommends “increased awareness of depression amongst U.S. Medical students” due to the increasing body of research describing the rise of depression, burn-out and suicide ideation among medical students. There is consequently a need to understand mental health issues as a component of professional development. Hypothesis: A student-led symposium addressing mental and emotional health topics relevant to medical students would reduce the stigma associated with mental illness. Materials and Methods: A 2-hour student-run “Patient Perspective” was held during the second neuroscience block at an osteopathic medical school in the northeastern United States. One week before the program, a student-developed online Wellness Survey measured prevalence of mental illness, common feelings during medical school, coping mechanisms used for stress, and use of mental health resources. Immediately before and after the program, students were asked to report their familiarity with mental illness and their feelings regarding a vignette about a mentally ill woman using “Mental Illness Among Us” pre and post surveys provided by the University of California San Francisco School of Medicine and adapted for the event. During the program, data from the online survey were shared, student organizers discussed emotional wellness and positive coping mechanisms in the context of the profession, and student panelists shared their experiences with mental health issues. A faculty psychiatrist spoke about mental health resources, and attendees received pamphlets listing these resources. The event concluded with student-led breakout sessions at which stress during medical school and strategies for promoting positive coping mechanisms were discussed, followed by the post survey. Results: 113 students completed the pre survey, 89 of whom completed the post survey. For these 89, differences between post and pre responses were universally in the direction of increasing acceptance and decreasing stigma of those with mental illness; all differences were statistically significant. The largest shift regarded students’ reluctance to disclose their own theoretical mental illness to colleagues. Conclusion: Incorporating an emotional health symposium into medical students’ training may increase understanding and acceptance of those who may have mental illness and reduce stigma associated with mental illness.https://digitalcommons.pcom.edu/posters/1002/thumbnail.jp

Herpes Simplex Virus 1 and Chlamydophila (Chlamydia) pneumoniae promote Ab 1-42 amyloid processing in murine astrocytes linking an infectious process to Alzheimer\u27s disease

Background: Several studies have suggested an infectious etiology for Alzheimer\u27s disease (AD). Previously, our laboratory identified Chlamydia pneumoniae (Cpn) from autopsied sporadic AD brains, as well as developed a BALB/c mouse model that demonstrated infection-induced amyloid plaques similar to those found in AD. Hypothesis: We propose that an additional pathogen such as herpes simplex virus type 1 (HSV1), also may be a contributing factor in toin the pathology seen in AD. HSV1, in addition to Cpn, may be triggering the abnormal cleavage of the beta amyloid precursor protein (bAPP) into Ab1-42 , thereby contributing to amyloid plaque formation. Our current study examines amyloid processing following infection of primary and C8-DIA murine astrocytes with Cpn and HSV1. Materials and Methods: Immunocytochemistry and western analysis was used to analyze the outcome of infection by these two pathogens. Results: Cpn infection resulted in an increase in cytoplasmic labeling of Ab 1-42 relative to uninfected cells, while increased nuclear labeling of Ab 1-42 was observed following HSV1 infection. Co-infections with Cpn and HSV1 resulted in amyloid labeling resembling that of HSV1 infection alone, though Ab 1-42 labeling appeared decreased specifically in Cpn-infected cells of the co-infected monolayers. Conclusions: These data suggest that infection of astrocytic cells by HSV1 and (Cpn) alter the processing of bAPP, thereby producing Ab1-42. Therefore, these studies, inaddition to the previous research reported by our laboratory, support an emerging linkage of the infectious processs to the neuropathology characteristic of Alzheimer\u27s disease.https://digitalcommons.pcom.edu/posters/1008/thumbnail.jp

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