This document summarizes research on viral promoter polymorphisms in HIV disease. It discusses how approximately 80% of HIV-infected individuals experience infection of the central nervous system, leading to neurological disorders. The research focuses on identifying single nucleotide polymorphisms (SNPs) in the HIV-1 long terminal repeat (LTR) that correlate with clinical parameters and neurological impairment. Nine SNPs were found to associate with changes in CD4 count and viral load. Further analysis found that these same SNPs identified in peripheral blood samples were also present in HIV-infected brain tissue. [END SUMMARY]
The document discusses the neurological effects of COVID-19. It begins by explaining that COVID-19 can lead to neurological problems in addition to being a respiratory illness. Some symptoms include loss of smell, headaches, fatigue, and brain fog. Studies have shown the virus can enter the brain and cause changes in brain metabolites. COVID-19 patients have shown reductions in N-acetyl-aspartate and elevations in choline and myo-inositol, indicating white matter abnormalities and damage from oxygen deprivation. The virus may directly infect the brain or induce injury indirectly through immune responses or hypoxia.
The document summarizes a study of 13 patients with herpes simplex encephalitis. All patients underwent clinical exams, CSF analysis, and MRI or CT scans of the brain. CSF analysis showed lymphocytic pleocytosis and elevated proteins in all patients, and PCR testing detected HSV-1 DNA in the CSF of all patients. The most common radiological feature was involvement of the temporal lobes, seen bilaterally in many patients. Some patients also showed lesions in other brain regions. All patients received acyclovir treatment, with outcomes to be presented later in the document.
This document provides an overview of HIV/AIDS, including:
- HIV is caused by the human immunodeficiency virus (HIV) which is a retrovirus.
- As of 2016, there were approximately 36.7 million people living with HIV globally.
- HIV diagnosis involves ELISA and Western blot tests to detect HIV antibodies and viral proteins.
- HIV treatment involves the use of antiretroviral drugs from several classes including nucleoside/nucleotide reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, protease inhibitors, fusion inhibitors, CCR5 co-receptor antagonists, and integrase inhibitors.
- Co-infections with tuberculosis require specialized treatment reg
Epigenetic Mechanisms Regulating the Cellular Response to HypoxiaMAASTRO clinic
Lecture by Prof. Lorenz Poellinger in the course: "Tumour Hypoxia: From Biology to Therapy III". For the complete e-Course see http://www.myhaikuclass.com/MaastroClinic/metoxia
This study found that the herpesvirus VP1/2 protein associates with the cellular dynein/dynactin microtubule motor complex and promotes retrograde transport of viral capsids. Specifically:
1) VP1/2 interacts with capsids through binding to the capsid protein pUL25. This binding "unmasks" VP1/2's ability to localize to the nuclear membrane and engage in microtubule-dependent transport.
2) VP1/2 is able to move itself and surrogate cargo (mitochondria) in a microtubule-dependent manner when not bound to capsids. This demonstrates VP1/2 can actively transport cargo.
3) VP1/2 directly interacts
Human Immunodeficiency Virus (HIV) infects cells by attaching to the CD4 receptor with its gp120 glycoprotein. It requires a co-receptor, CCR5 or CXCR4, for fusion and entry into the cell. Once inside, the viral RNA is converted to DNA by reverse transcriptase. The DNA integrates into the host genome with the help of integrase. The virus hijacks the cell's machinery to produce new viral proteins and RNA, which are assembled into new virus particles that bud from the cell.
Slideshow is from the University of Michigan Medical
School's M1 Infectious Disease / Microbiology sequence
View additional course materials on Open.Michigan:
openmi.ch/med-M1IDM
The document discusses the neurological effects of COVID-19. It begins by explaining that COVID-19 can lead to neurological problems in addition to being a respiratory illness. Some symptoms include loss of smell, headaches, fatigue, and brain fog. Studies have shown the virus can enter the brain and cause changes in brain metabolites. COVID-19 patients have shown reductions in N-acetyl-aspartate and elevations in choline and myo-inositol, indicating white matter abnormalities and damage from oxygen deprivation. The virus may directly infect the brain or induce injury indirectly through immune responses or hypoxia.
The document summarizes a study of 13 patients with herpes simplex encephalitis. All patients underwent clinical exams, CSF analysis, and MRI or CT scans of the brain. CSF analysis showed lymphocytic pleocytosis and elevated proteins in all patients, and PCR testing detected HSV-1 DNA in the CSF of all patients. The most common radiological feature was involvement of the temporal lobes, seen bilaterally in many patients. Some patients also showed lesions in other brain regions. All patients received acyclovir treatment, with outcomes to be presented later in the document.
This document provides an overview of HIV/AIDS, including:
- HIV is caused by the human immunodeficiency virus (HIV) which is a retrovirus.
- As of 2016, there were approximately 36.7 million people living with HIV globally.
- HIV diagnosis involves ELISA and Western blot tests to detect HIV antibodies and viral proteins.
- HIV treatment involves the use of antiretroviral drugs from several classes including nucleoside/nucleotide reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, protease inhibitors, fusion inhibitors, CCR5 co-receptor antagonists, and integrase inhibitors.
- Co-infections with tuberculosis require specialized treatment reg
Epigenetic Mechanisms Regulating the Cellular Response to HypoxiaMAASTRO clinic
Lecture by Prof. Lorenz Poellinger in the course: "Tumour Hypoxia: From Biology to Therapy III". For the complete e-Course see http://www.myhaikuclass.com/MaastroClinic/metoxia
This study found that the herpesvirus VP1/2 protein associates with the cellular dynein/dynactin microtubule motor complex and promotes retrograde transport of viral capsids. Specifically:
1) VP1/2 interacts with capsids through binding to the capsid protein pUL25. This binding "unmasks" VP1/2's ability to localize to the nuclear membrane and engage in microtubule-dependent transport.
2) VP1/2 is able to move itself and surrogate cargo (mitochondria) in a microtubule-dependent manner when not bound to capsids. This demonstrates VP1/2 can actively transport cargo.
3) VP1/2 directly interacts
Human Immunodeficiency Virus (HIV) infects cells by attaching to the CD4 receptor with its gp120 glycoprotein. It requires a co-receptor, CCR5 or CXCR4, for fusion and entry into the cell. Once inside, the viral RNA is converted to DNA by reverse transcriptase. The DNA integrates into the host genome with the help of integrase. The virus hijacks the cell's machinery to produce new viral proteins and RNA, which are assembled into new virus particles that bud from the cell.
Slideshow is from the University of Michigan Medical
School's M1 Infectious Disease / Microbiology sequence
View additional course materials on Open.Michigan:
openmi.ch/med-M1IDM
HIV is a lentivirus which can not only infect actively dividing cells but also non-dividing cells such as macrophages. AIDS is the last stage of HIV infection. HIV primarily attacks T- helper cells resulting into low activated T-cytotoxic cells and suppression of immune system. thus leading to AIDS.
This document discusses the neuropsychiatric manifestations of COVID-19. It notes that SARS-CoV-2 can invade the brain through direct routes like hematogenous spread or neural pathways, or indirectly through immune activation and cytokine release. Common neuropsychiatric symptoms reported in COVID-19 patients include headaches, dizziness, impaired consciousness, strokes, and psychiatric conditions like depression and psychosis. The document explores several mechanisms by which the virus may cause neuroinflammation and neuroinvasion, resulting in neuropsychiatric complications.
This document provides information on the phenotype and genotype of COVID-19. It discusses that coronaviruses are RNA viruses that cause respiratory infections in humans. COVID-19 is caused by a novel betacoronavirus. The virus particle is spherical with spikes and enters cells by binding to the ACE2 receptor. It contains a positive-sense RNA genome that is translated and replicated via a replicase-transcriptase complex to make structural proteins and more virus particles.
This document provides an outline for a presentation on HCV genotyping methods. It discusses the morphology and characteristics of HCV, its history and structure. It describes the six genotypes of HCV and their geographical distribution. It also discusses mutants of HCV, clinical importance of genotypes, laboratory diagnosis, and different methods for genotyping including direct sequencing, RFLP typing of the 5'UTR, and line probe assay (LiPA). The advantages of LiPA include it being easy, less time consuming and capable of reliably genotyping HCV RNA directly from clinical samples.
this presentation covers the detailed information about the non-infectious meningitis and its pathology, epidemiology, causes, mechanism and its clear pathophysiology. have a glance to know more about it . thank you...
This thesis investigates the role of defective interfering particles (DIs) in the induction of interferon-β (IFN-β) following infection with parainfluenza virus 5 (PIV5). Using A549 reporter cells containing a GFP gene under control of the IFN-β promoter, the author shows that only a limited number of infected cells induce IFN-β, demonstrating a heterocellular response. Knockdown of RIG-I, MDA5 and LGP2 is used to determine the pattern recognition receptors involved in IFN induction by paramyxoviruses. Results support previous findings that DIs generated during errors in viral replication, rather than intact viral genomes, are the primary pathogen associated molecular patterns
Summerlin Asset Management seeks to purchase bank-owned real estate portfolios at a steep discount. They will acquire first-position mortgage notes and deeds of trust secured by residential properties. Their goal is to service these assets and make the mortgages performing again, then resell them at a profit. This provides investors an opportunity to invest in discounted mortgages and real estate notes through Summerlin. They have relationships with major banks to purchase large portfolios of non-performing loans.
This document summarizes research being conducted on the neurological consequences of HIV infection. It discusses how:
1) HIV infection of the central nervous system occurs in approximately 80% of infected individuals and around 50% will experience some form of neurological disorder.
2) While antiretroviral therapy has reduced some HIV-associated neurocognitive disorders, their prevalence is increasing due to improved survival of HIV patients.
3) The research aims to investigate how specific HIV-1 long terminal repeat single nucleotide polymorphisms in patient blood samples correlate with alterations in clinical indicators of HIV disease progression in the era of antiretroviral therapy.
The document outlines Erik Erikson's 5 stages of psychosocial development, including Basic Trust vs. Mistrust in infancy, Autonomy vs. Shame and Doubt in early childhood, Initiative vs. Guilt in preschool age, Industry vs. Inferiority in school age children, and Identity vs. Role Confusion in adolescence. It provides a brief description of each stage and the potential psychological impacts of not developing properly during each respective life phase. The document appears to be class notes outlining Erikson's theory of psychosocial development for EDTE 314 taught by Dr. Love during spring 2013.
This document discusses the challenges of disconnected work activities and tools, including random work requests, poor visibility of work and resources, and inefficient use of time. It promotes a single system of engagement solution that provides total visibility, efficiency, and productivity through connected tools and activities. The solution is said to help reduce costs, improve relationships between teams, and allow organizations to see what's happening and plan accordingly.
This document provides an overview of becoming an agent for Summerlin Asset Management (SAM) to sell real estate investment notes. It outlines the benefits of being a SAM agent including residual income from commissions with minimal work. The note buying process involves SAM purchasing notes from banks and reselling them to investors. Marketing involves using SAM's website and materials to connect agents directly to investors via calls or meetings. The document addresses common investor objections and provides responses, and outlines the next steps an agent would take to set up a note purchase for an investor through SAM.
This document discusses rabies virus, the immune response to rabies, and rabies vaccines. It first provides background on rabies virus, describing its structure, life cycle, and pathogenesis. It then examines the immune response to rabies in the periphery and central nervous system, highlighting both the expected immune response and viral evasion mechanisms. Finally, it reviews current rabies vaccines for humans and animals as well as new vaccine approaches under development.
A detailed description of HIV covering virology, morphology, pathogenesis, clinical stages and manifestations, laboratory diagnosis, and diagnostic strategy, and therapeutic options and prevention.
This document provides an overview of Hepatitis C. It begins with an introduction stating that over 71 million people worldwide are chronically infected with HCV. It then covers the virology of HCV including its structure, genome, replication cycle, genotypes/quasispecies. The epidemiology section discusses the global prevalence and incidence. Pathogenesis outlines how HCV evades the immune system to cause chronic infection. Clinical features are separated into acute hepatitis C and chronic hepatitis C. Extrahepatic manifestations associated with HCV are also summarized.
A LECTURE ON AIDS FOR FIRST MBBS STUDENTS, DEPT OF BIOCHEMISTRY.
A CLASS ON EPIDEMIOLOGY, VIROLOGY,HIV-MORPHOLOGY, GENOME, LIFE CYCLE,MODE OF TRANSMISSION, IMMUNOLOGY, PATHOPHYSIOLOGY AND PATHOGENESIS, LABORATORY DIAGNOSIS AND MANAGEMENT.
HIV infection
Mode of transmission, pathogenesis, clinical manifestations, laboratory diagnosis, treatment, prevention, prognosis, scope of AIDS vaccine.
Fifth Annual Mitchell Memorial Lecture, October 6, 2014, at UC San Diego, featuring Dr. Jonathan Karn of Case Western Reserve University speaking on "Lessons Learned from models for HIV latency helping to formulate virus eradication strategies."
This document summarizes HIV and retroviruses. It discusses that retroviruses possess reverse transcriptase which converts viral RNA to DNA. HIV is classified as a lentivirus that causes AIDS. HIV was discovered in 1983 and is the causative agent of AIDS. It is a retrovirus with an RNA genome and envelope. The virus enters host cells using envelope proteins gp120 and gp41 to bind CD4 receptors. The virus has structural and regulatory genes. Infection progresses from primary infection to clinical latency to AIDS as CD4 counts decline. Opportunistic infections occur when CD4 counts drop below 200. Common infections include PCP, CMV, TB, candidiasis and cancers like Kaposi's sarcoma.
HIV is a lentivirus which can not only infect actively dividing cells but also non-dividing cells such as macrophages. AIDS is the last stage of HIV infection. HIV primarily attacks T- helper cells resulting into low activated T-cytotoxic cells and suppression of immune system. thus leading to AIDS.
This document discusses the neuropsychiatric manifestations of COVID-19. It notes that SARS-CoV-2 can invade the brain through direct routes like hematogenous spread or neural pathways, or indirectly through immune activation and cytokine release. Common neuropsychiatric symptoms reported in COVID-19 patients include headaches, dizziness, impaired consciousness, strokes, and psychiatric conditions like depression and psychosis. The document explores several mechanisms by which the virus may cause neuroinflammation and neuroinvasion, resulting in neuropsychiatric complications.
This document provides information on the phenotype and genotype of COVID-19. It discusses that coronaviruses are RNA viruses that cause respiratory infections in humans. COVID-19 is caused by a novel betacoronavirus. The virus particle is spherical with spikes and enters cells by binding to the ACE2 receptor. It contains a positive-sense RNA genome that is translated and replicated via a replicase-transcriptase complex to make structural proteins and more virus particles.
This document provides an outline for a presentation on HCV genotyping methods. It discusses the morphology and characteristics of HCV, its history and structure. It describes the six genotypes of HCV and their geographical distribution. It also discusses mutants of HCV, clinical importance of genotypes, laboratory diagnosis, and different methods for genotyping including direct sequencing, RFLP typing of the 5'UTR, and line probe assay (LiPA). The advantages of LiPA include it being easy, less time consuming and capable of reliably genotyping HCV RNA directly from clinical samples.
this presentation covers the detailed information about the non-infectious meningitis and its pathology, epidemiology, causes, mechanism and its clear pathophysiology. have a glance to know more about it . thank you...
This thesis investigates the role of defective interfering particles (DIs) in the induction of interferon-β (IFN-β) following infection with parainfluenza virus 5 (PIV5). Using A549 reporter cells containing a GFP gene under control of the IFN-β promoter, the author shows that only a limited number of infected cells induce IFN-β, demonstrating a heterocellular response. Knockdown of RIG-I, MDA5 and LGP2 is used to determine the pattern recognition receptors involved in IFN induction by paramyxoviruses. Results support previous findings that DIs generated during errors in viral replication, rather than intact viral genomes, are the primary pathogen associated molecular patterns
Summerlin Asset Management seeks to purchase bank-owned real estate portfolios at a steep discount. They will acquire first-position mortgage notes and deeds of trust secured by residential properties. Their goal is to service these assets and make the mortgages performing again, then resell them at a profit. This provides investors an opportunity to invest in discounted mortgages and real estate notes through Summerlin. They have relationships with major banks to purchase large portfolios of non-performing loans.
This document summarizes research being conducted on the neurological consequences of HIV infection. It discusses how:
1) HIV infection of the central nervous system occurs in approximately 80% of infected individuals and around 50% will experience some form of neurological disorder.
2) While antiretroviral therapy has reduced some HIV-associated neurocognitive disorders, their prevalence is increasing due to improved survival of HIV patients.
3) The research aims to investigate how specific HIV-1 long terminal repeat single nucleotide polymorphisms in patient blood samples correlate with alterations in clinical indicators of HIV disease progression in the era of antiretroviral therapy.
The document outlines Erik Erikson's 5 stages of psychosocial development, including Basic Trust vs. Mistrust in infancy, Autonomy vs. Shame and Doubt in early childhood, Initiative vs. Guilt in preschool age, Industry vs. Inferiority in school age children, and Identity vs. Role Confusion in adolescence. It provides a brief description of each stage and the potential psychological impacts of not developing properly during each respective life phase. The document appears to be class notes outlining Erikson's theory of psychosocial development for EDTE 314 taught by Dr. Love during spring 2013.
This document discusses the challenges of disconnected work activities and tools, including random work requests, poor visibility of work and resources, and inefficient use of time. It promotes a single system of engagement solution that provides total visibility, efficiency, and productivity through connected tools and activities. The solution is said to help reduce costs, improve relationships between teams, and allow organizations to see what's happening and plan accordingly.
This document provides an overview of becoming an agent for Summerlin Asset Management (SAM) to sell real estate investment notes. It outlines the benefits of being a SAM agent including residual income from commissions with minimal work. The note buying process involves SAM purchasing notes from banks and reselling them to investors. Marketing involves using SAM's website and materials to connect agents directly to investors via calls or meetings. The document addresses common investor objections and provides responses, and outlines the next steps an agent would take to set up a note purchase for an investor through SAM.
This document discusses rabies virus, the immune response to rabies, and rabies vaccines. It first provides background on rabies virus, describing its structure, life cycle, and pathogenesis. It then examines the immune response to rabies in the periphery and central nervous system, highlighting both the expected immune response and viral evasion mechanisms. Finally, it reviews current rabies vaccines for humans and animals as well as new vaccine approaches under development.
A detailed description of HIV covering virology, morphology, pathogenesis, clinical stages and manifestations, laboratory diagnosis, and diagnostic strategy, and therapeutic options and prevention.
This document provides an overview of Hepatitis C. It begins with an introduction stating that over 71 million people worldwide are chronically infected with HCV. It then covers the virology of HCV including its structure, genome, replication cycle, genotypes/quasispecies. The epidemiology section discusses the global prevalence and incidence. Pathogenesis outlines how HCV evades the immune system to cause chronic infection. Clinical features are separated into acute hepatitis C and chronic hepatitis C. Extrahepatic manifestations associated with HCV are also summarized.
A LECTURE ON AIDS FOR FIRST MBBS STUDENTS, DEPT OF BIOCHEMISTRY.
A CLASS ON EPIDEMIOLOGY, VIROLOGY,HIV-MORPHOLOGY, GENOME, LIFE CYCLE,MODE OF TRANSMISSION, IMMUNOLOGY, PATHOPHYSIOLOGY AND PATHOGENESIS, LABORATORY DIAGNOSIS AND MANAGEMENT.
HIV infection
Mode of transmission, pathogenesis, clinical manifestations, laboratory diagnosis, treatment, prevention, prognosis, scope of AIDS vaccine.
Fifth Annual Mitchell Memorial Lecture, October 6, 2014, at UC San Diego, featuring Dr. Jonathan Karn of Case Western Reserve University speaking on "Lessons Learned from models for HIV latency helping to formulate virus eradication strategies."
This document summarizes HIV and retroviruses. It discusses that retroviruses possess reverse transcriptase which converts viral RNA to DNA. HIV is classified as a lentivirus that causes AIDS. HIV was discovered in 1983 and is the causative agent of AIDS. It is a retrovirus with an RNA genome and envelope. The virus enters host cells using envelope proteins gp120 and gp41 to bind CD4 receptors. The virus has structural and regulatory genes. Infection progresses from primary infection to clinical latency to AIDS as CD4 counts decline. Opportunistic infections occur when CD4 counts drop below 200. Common infections include PCP, CMV, TB, candidiasis and cancers like Kaposi's sarcoma.
This document summarizes key information about HIV/AIDS, including its history, virology, diagnosis, treatment, and prevention. It describes how HIV was first identified in 1981 as the cause of AIDS, belongs to the retrovirus family, and has two types, HIV-1 and HIV-2. Over 30 million people have died of AIDS since 1981, and approximately 2.5 million people are newly infected with HIV each year.
This document provides an overview of HIV/AIDS, including its classification, transmission, epidemiology, virology, pathogenesis, and treatment. Key points include:
- HIV is a lentivirus that infects CD4+ cells and integrates into the host genome. It has high genetic diversity and can lie dormant for years.
- HIV is transmitted sexually, through blood exposure, or mother-to-child. Factors like behavior patterns, condom use, and availability of treatment impact spread.
- HIV evades the immune system through rapid mutation, hiding in sanctuaries, and immune activation leading to exhaustion. This allows the virus to persist despite immune responses.
- HIV causes
This document discusses immunodeficiency disorders. It describes how deficiencies in antibodies, phagocytes, or complement components can lead to infections by extracellular bacteria. Deficiencies in cell-mediated immunity are associated with recurrent viral, protozoal, and fungal infections. Primary immunodeficiencies are caused by genetic defects, while secondary immunodeficiencies result from infection, toxicity, radiation, or other acquired factors. Specific primary immunodeficiencies that affect B cells, T cells, phagocytes, or the complement system are described. The document also discusses acquired immunodeficiency syndrome (AIDS) caused by HIV, outlining the virus structure, pathogenesis, clinical features, and diagnosis.
Structure of Virus, modes of transmission, pathogenesis, clinical features, biochemical basis of clinical symptoms, laboratory diagnosis, treatment and prevention.
- Retroviridae are a family of enveloped RNA viruses that include HIV and have been associated with cancers and AIDS. HIV is classified as a lentivirus within this family.
- There are two main theories for the origin of HIV in humans - natural transfer from chimpanzees through hunting or a contaminated oral polio vaccine.
- HIV infects CD4+ T cells using its envelope proteins to bind host receptors, then integrates into the host genome and uses host cell machinery to replicate. This evades immune detection and allows the virus to persist.
Human immunodeficiency virus (HIV) is a retrovirus that causes acquired immunodeficiency syndrome (AIDS). HIV infects and kills CD4+ T cells, leading to a weakened immune system. There are two types of HIV - HIV-1 and HIV-2. HIV-1 is more virulent and prevalent globally. HIV enters cells by binding to CD4 receptors and either CCR5 or CXCR4 coreceptors and fusing with the cell membrane. Inside the cell, HIV converts its RNA to DNA using reverse transcriptase and integrates into the host cell's DNA.
JC Virus of the CNS classically presents as progressive multifocal leukoencephalopathy, but on rare occasion can manifest as septic meningitis. Slides compares the presentation, workup and treatment in both forms.
The document summarizes key information about HIV/AIDS, including:
- HIV is transmitted sexually, through shared needles, or mother-to-child. It causes AIDS by destroying CD4 cells.
- The disease was first recognized in 1981 in the US. The virus was isolated in 1983-1984.
- High risk groups for HIV infection include men who have sex with men, intravenous drug users, and heterosexual contact.
- HIV progresses from acute infection to asymptomatic latency to full-blown AIDS as CD4 cell counts decline below 200.
- Opportunistic infections define AIDS as the immune system is compromised.
- Diagnosis involves detecting antibodies or viral components. Treatment aims to suppress viral
Human Retroviruses are RNA viruses that contain the enzyme reverse transcriptase, allowing them to convert their RNA genome into DNA. The two major genera that affect humans are Lentiviruses, which include HIV-1 and HIV-2, and HTLV-BLV group, which includes HTLV-1 and HTLV-2. HIV binds host cells via gp120, enters via fusion, reverse transcribes into DNA then integrates into the host genome. It replicates using host cell machinery. Infection can lead to AIDS as CD4+ T cells are depleted. Opportunistic infections are treated with antiretrovirals that target reverse transcriptase and protease.
This document discusses the epidemiological features and modes of transmission of HIV. It describes the virus itself, including that it is an enveloped retrovirus with two RNA strands. It spreads primarily through sexual contact, blood transfusions, needle sharing, and from mother to child. The incubation period from infection to AIDS symptoms is highly variable, ranging from less than a year to over 15 years. Key host factors are age 20-49 years and belonging to high-risk groups like men who have sex with men.
This document provides an overview of recent advances in neurotropic viruses. It begins with definitions of neurotropic viruses and examples of how they can present clinically. It then lists many common and less common neurotropic viruses, including herpesviruses, enteroviruses, arboviruses, and retroviruses. The document discusses epidemiology, pathogenesis, laboratory diagnosis including newer methods like PCR and serology, specific viruses like HSV, CMV, enteroviruses and poliovirus. It covers diagnosis, treatment and vaccination strategies for these neurotropic viruses.
1. Viral Promoter Polymorphisms
in HIV Disease
Gregory C. Antell
2013 Sigma Xi Research Showcase
March 15, 2013
Graduate Student
Drexel University
School of Biomedical Engineering, Science, and Health Systems
2. The HIV epidemic has neurological consequences
An average of 6,800 new HIV infections and 5,700 HIV-related
deaths occur daily worldwide
Infection of the central nervous system occurs in approximately
80% of infected individuals
Approximately 50% of HIV-infected adults and children will
demonstrate a neurological disorder at one time
The advent of anti-retroviral therapy has diminished the incidence
of HIV-associated neurocognitive disorders to a lesser extent than
other AIDS-related diseases
Prevalence of neurocognitive diseases has actually increased
due to the prolonged survival of HIV infected individuals
Neuropathology of HIV disease remains largely unknown and a
critical area of current and future research
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
3. Multiple factors influence HIV-1 pathogenesis and HIV-1-
associated neurocognitive disorders (HAND)
Host & Therapy Cellular
Pathogenesis is shaped A spectrum of cellular targets are
by the host immune vulnerable to infection, which may lead to
response and genetics, physiological compartmentalization and
drug therapy, drug tissue-specific selective pressures.
abuse, and aging.
HAND
Viral
Molecular diversity emerges in the
virus as it adapts to selective HIV-associated neurocognitive
pressures. Particular variants may disorders include HIV-associated
serve as biomarkers of advanced dementia (HAD) and minor
neurological disease. cognitive motor disorder (MCMD).
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
4. HIV-1 pathogenesis and associated neurological dysfunction
CNS viral evolution Brain
Blood
Brain
Barrier
CNS viral entry
Extra-CNS viral evolution Blood
HIV-1 likely enters the brain during acute infection and during the absence
of effective therapy or immune dysfunction
In the brain resident microglial cells and perivascular macrophages are the
major cellular targets for infection
Release of viral and cellular neurotoxic mediators results in the alteration of
the blood-brain barrier and neuronal dysfunction
Acute Infection Clinical Latency AIDS / Dementia
CD4 count > 500 200-500 < 200
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
5. HIV-1 CNS entry and infection of resident cell populations
Peripheral Blood Brain
Mucosal compartment HIV-1-infected
perivascular
macrophage
Lymphoid compartment
resident
microglia
cells
Bone marrow compartment Viral gene products have
neurotoxic effects on
astrocytes and neurons
Blood-
Other end organs Brain
Barrier astrocytes neurons
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
6. HIV-1 replication scheme
HIV-1 replication is controlled by the viral promoter, termed the long terminal repeat (LTR), as
well as the regulatory genes Tat and Vpr
While HIV-1 is known to have an entry phenotype, it is hypothesized that it may also have
distinct replication phenotypes that associates with particular host cell phenotypes and/or
physiological compartments
Absorption and entry
gp120
CD4 Budding
Coreceptors
CCR5
CXCR4 genomic Assembly
Reverse RNA
Transcription
Protein synthesis
Nuclear Transport Viral Gene and processing
and Integration Expression
integrated
proviral DNA mRNA
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
7. The HIV-1 genome and 5’-LTR organization
tat
vpr
rev
5’LTR gag vif 3’LTR
pol vpu env nef
455 552 638
nt1 leader
U3 R U5
nuc-0 HS2 HS3
+1
nuc-1 HS4
-405 -245 +20 +165
Modulatory region Core region
Enhancer region
The HIV-1 genome is flanked by two LTR sequences: the 5’-LTR and the 3’-LTR
The 5’-LTR acts as the promoter for viral gene expression
The LTR contains a high concentration of binding sites for cellular transcription factors, which
can vary according to the host cell phenotype
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
8. Background and demographics of the DREXELMED HIV/AIDS
Genetic Analysis Cohort
Patients enrolled in the DrexelMed Cohort are recruited from the Patients
Philadelphia region and are scheduled to return every six months. Visit
Seen
At each visit, a patient interview is conducted, a blood sample is
collected, and a neurological exam is performed. Initial Visit 503
First Return 298
Demographic Count (%)/Mean (+/- SD) Second Return 202
Categories
Variables with clinical variables
Male 332 (66%) Third Return 136
Gender
Female 169 (33.6%) Fourth Return 95
Black/AA 418 (83.1%)
Fifth Return 67
White 62 (12.3%)
Race Other (AI/AN, multiple, Sixth Return 43
16 (3.2%)
asian)
Seventh Return 29
Unknown 7 (1.4%)
cH 424 (84.3%) Eighth Return 17
HAART status dH 43 (8.5%) Ninth Return 7
nH 34 (6.7%)
Tenth Return 2
Age 45.43 (± 8.569)
Years since diagnosed 11.916 (± 7.312) Total 1399
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
9. Research Focus #1:
Do specific HIV-1 LTR single nucleotide
polymorphisms (SNPs) derived patient
peripheral blood samples correlate with
alterations in clinical HIV disease
parameters in the HAART era?
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
10. Viromic analysis of DREXELMED HIV/AIDS Genetic Analysis
Cohort in the HAART era
Ficoll-Pacque Qiagen DNEasy
Whole Blood Plus gradient Tissue Kit
Serum & PBMC
separation Luminex Human PCR amplify
BSL-3 Facility Cytokine 30 plex proviral DNA
Separate on
Serum and cell banking agarose gel
Clinical and virus/host genomic data management
HIV-1 Sequence Database
Sequence PCR product
analysis sequencing
Gel extraction
pGL3 Basic pCR4-TOPO
Incubate with
PCR amplify/ Taq to add A
Functional overhang
analysis clone proviral DNA
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
11. HIV-1 LTR SNP densities in patients from the DREXELMED
HIV/AIDS Genetic Analysis Cohort
900 700
Sequence Coverage
800
SNP Density (Number of mutations)
SNP Density 600
Coverage (Number of Sequences)
700
500
600
500 400
400 300
300
200
200
100
100
0 0
115
381
400
134
153
172
191
210
229
248
267
286
305
324
343
362
419
438
457
476
495
514
533
552
571
590
609
628
20
39
58
77
96
1
Nucleotide Position on ConB (Jan 2002) Reference Sequence
• LTR SNP coverage and frequency was calculated for 461 patients and 1127
sequences
• SNPs are observed throughout the LTR sequence and can be mapped to
transcription factor binding sites
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
12. Nine HIV-1 LTR SNPs associate with change in CD4 count and log
viral load away from the average of the cohort
The single nucleotide polymorphisms (SNPs) identified from patient peripheral
blood samples can be plotted according to base pair position in the LTR and
association with CD4+ T cell count and log viral load
Data is adjusted for patient age, sex, and race
INSTITUTE FOR MOLECULAR MEDICINE AND INFECTIOUS DISEASE
13. Significant LTR SNPs
Phenotype Position Ref./Mut. Mutant Freq Effect p-value
108 A/CGT 38.0% -41.228 0.0176
120 C/AT 6.2% 72.950 0.0200
CD4 Count
181 A/CG 8.3% -72.320 0.0173
293 G/ACT 11.4% -46.920 0.0452
108 A/CGT 38.0% 184.4% 0.0010
115 A/GT 18.5% 60.7% 0.0301
Viral Load 160 C/AG 6.3% 46.7% 0.0278
168 G/ACT 14.8% 60.2% 0.0282
251 G/ACT 8.8% 53.9% 0.0315
A total of 9 SNPs, located at 8 distinct nucleotide positions, were identified to associate
with the clinical parameters of CD4+ T cell count and/or viral load at a statistically
significant level (p-vale < 0.05). The effect in this case is defined as the change in
CD4+ T cell count or the percent change in viral load away from the average.
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14. Research Focus #2:
Are these significant peripheral blood
HIV-1 LTR single nucleotide
polymorphisms (SNPs) also found in
HIV-infected brains?
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15. Isolation of HIV-1 brain-derived LTRs for sequence analysis
Nested PCR amplifies LTR
Autopsy tissue punches QIAGEN DNeasy
from proviral DNA
Tissue Procedure
PCR
Isolation of amplify
genomic DNA proviral
DNA
HIV-1 Brain LTR Sequence Database Separate on agarose gel
Preparation for
sequencing and
sequence analysis
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16. Number of brain and spleen sequences used in analysis
Patient samples available Brain
HAD 6 Normal Subsyndromic MCMD HAD
MCMD 16 Sequence
Subsyndromic 3 Number 16 18 95 38
Normal 2 Patient
Uninfected 1 Number 2 2 14 6
TOTAL 28 Spleen
Tissue regions available Normal Subsyndromic MCMD HAD
Cerebellum 28 Sequence
Deep White Matter 28 Number 3 2 19 7
Head of Caudate 28 Patient
Midfrontal Gyrus 28 Number 2 2 14 4
Parietal 28
Thalamus 27 Autopsy tissue samples were collected from multiple
Spleen 23 brain sites, as well as spleen, from patients with
varying degrees of neurological impairment.
National NeuroAIDS Tissue Consortium – University of Texas
Director: Ben Gelman, M.D., Ph.D.
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17. Prevalence of clinically significant peripheral blood LTR SNPs
in HIV-1 infected brain tissue
Nucleotide TF Number of Total in Total in Neuro. Neuro.
Position Site individuals Spleen Brain Normal Impaired
COUP/
108 20 11 75 13 73
AP1
COUP/
115 5 1 6 0 7
AP1
120 COUP 5 1 10 1 10
160 AP1 2 0 3 0 3
168 unk 8 1 10 0 11
181 unk 4 4 0 0 4
251 unk 10 4 13 0 17
293 USF 8 5 8 0 13
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18. Clinically significant peripheral blood HIV-1 LTR SNPs are found in
all regions of the HIV-1-infected brain except for SNP 181
16
14
12
Number of SNPs
Cerebellum
10
Deep White Matter
8 Head of Caudate
Midfrontal Gyrus
6
Parietal
4 Thalamus
2
0
108 115 120 160 168 181 251 293
LTR SNP Position
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19. HIV-1 LTR SNPs identified in the peripheral blood are also
found to associate with neurologic impairment in the brain
Nucleotide Found in
TF Site Texas Cohort Notes DREXELMED PBMC Notes
Position Brain?
Decreases with Decreased CD4 count
108 Yes COUP/AP1
impairment Increased viral load
115 Yes COUP/AP1 Only in impairment Increased viral load
120 Yes COUP Mostly in impairment Increased CD4 count
Rare, only found in
160 Yes AP1 Increased viral load
brain and impairment
168 Yes unk Only in impairment Increased viral load
181 No unk Only found in spleen Decreased CD4 count
251 Yes unk Only in impairment Increased viral load
293 Yes USF Only in impairment Decreased CD4 count
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20. Frequency of LTR position 108 polymorphism (A to G) with
respect to neurocognitive status
100% 92% 100% 90%
90%
80%
70% 63%
60% 54%
50% 50% BRAIN
50% 46%
40% SPLEEN
30%
20%
10%
0%
NORMAL SUBSYNDROMIC MCMD HAD
BRAIN SPLEEN
Nucleotide Normal Subsyndromic MCMD HAD Nucleotide Normal Subsyndromic MCMD HAD
A (reference) 1 1 24 13 A (reference) 0 1 7 2
G (mutation) 11 9 40 15 G (mutation) 2 1 6 2
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21. Frequency of LTR position 168 polymorphism (G to A) with
respect to neurocognitive status
25%
20% 18%
15%
10% 8% 8%
5%
0% 0% 0% 0% 0% BRAIN
0%
SPLEEN
BRAIN SPLEEN
Normal Subsyndromic MCMD HAD Normal Subsyndromic MCMD HAD
G (reference) 12 10 54 24 G (reference) 2 2 10 4
A (mutation) 0 0 10 4 A (mutation) 0 0 3 0
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22. Frequency of LTR position 251 polymorphism (G to A/C) with
respect to neurocognitive status
25% 23%
20%
16%
15% 14%
BRAIN
10%
SPLEEN
5%
0% 0% 0% 0% 0%
0%
NORMAL SUBSYNDROMIC MCMD HAD
BRAIN SPLEEN
Normal Subsyndromic MCMD HAD Normal Subsyndromic MCMD HAD
G (reference) 12 10 54 24 G (reference) 2 2 10 4
A/C (mutation) 0 0 10 4 A/C (mutation) 0 0 3 0
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23. Brain-derived HIV-1 LTR vSNPs at positions 115, 120, 160,
and 293 associated with neurocognitive impairment
50% 50%
SNP 115 SNP 120
40% 40%
A to G/T C to T
30% 30%
25%
20% 20%
11% 11%
8% 8% 7%
10% 5%
10%
0% 0% 0% 0% 0% 0% 0% 0% 0%
0% 0%
NORMAL SUBSYNDROMIC MCMD HAD NORMAL SUBSYNDROMIC MCMD HAD
50% 50%
SNP 160 SNP 293 38%
40% 40%
C to A/G G to A/C
30% 30%
20% 20%
13%
10% 5% 10%
0% 0% 0% 0% 0% 0% 0% 0% 0% 0% 0% 0% 0%
0% 0%
NORMAL SUBSYNDROMIC MCMD HAD NORMAL SUBSYNDROMIC MCMD HAD
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24. Summary of major findings
• Eight HIV-1 LTR SNPs derived from peripheral blood
mononuclear cells associate with change in CD4 count and/or
log viral load away from the average of the cohort
• Clinically significant peripheral blood HIV-1 LTR SNPs are found
in all regions of the HIV-1-infected brain except for SNP 181
• HIV-1 LTR SNPs identified in the peripheral blood are also
found to associate with neurologic impairment in the brain,
particularly SNPs 108, 168, and 251
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25. Future directions
• Identify SNPs in PBMC-derived LTR sequence that correlate
with neurological disease and determine if they are present in
HIV-1-infected brain tissue
• Identify SNPs in brain-derived LTR sequences that associate
with neurological impairment, and assess their presence in
PBMC-derived LTRs
• Analyze additional HIV genes that contribute to proviral
transcription, such as Tat and Vpr, for single nucleotide
polymorphisms that correlate with clinical parameters
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26. Ultimate objective of this research
To identify a panel of genetic variants in the
proviral HIV-1 LTR (or other parts of the genome)
derived from PBMCs that are predictive of
neurologic decline
We envision a scenario where a simple blood test and diagnostic
PCR can cue physicians about potential problems and treatment
strategies. This viral SNP marker panel would be used in tandem
with other neurocognitive biomarkers.
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27. Brian Wigdahl, Ph.D., Professor & Chair
Department of Microbiology & Immunology
Drexel University College of Medicine
Director
William Dampier, Ph.D. Olimpia Meucci, M.D., Ph.D. Betty Condran Renzo Perales
Rui Feng, Ph.D. Sonia Navas-Martin, Ph.D. Jessica Cross Matt Rimbey
Jeffrey Jacobson, M.D. Michael Nonnemacher, Ph.D. Satinder Dahiya Germaine Rival
Pooja Jain, Ph.D. Vanessa Pirrone, Ph.D. David Downie Fiorella Rossi
Steve Jennings, Ph.D. Laura Steel, Ph.D. Brian Frantz Sonia Shah
Zafar Khan, Ph.D. Nirzari Parikh, M.S. Archana Gupta Luz Jeanette Sierra
Sandhya Kortagere, Ph.D. Shendra Passic, M.S. Nneka Ikpeze Marianne Strazza
Fred Krebs, Ph.D. Benjamas Aiamkitsumrit Shawn Keogan Gokul Swaminathan
Michele Kutzler, Ph.D. Greg Antell Christina Kollias Ken Thompson
David Libon, M.D. Brandon Blakey Sharon Lewis Cristian Valencia
Julio Martin-Garcia, Ph.D. Jessica Brown Raphael Lukov Jean Williams
Brian Moldover, Ph.D. Natalie Chen Andrea Partridge Wen Zhong
NINDS NIMH NCI NIDA NIAID
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Editor's Notes
1. Thinking about HIV: the intersection of virus, neuroinflammation and cognitive dysfunction2. Immune privilege and HIV-1 persistence in the CNS
So to address our question, we need to isolate viral sequences from our patients and compare them to clinical parameters indicative of disease progression. To do this we draw whole blood from patients participating in our cohort, which then enters our BSL-3 facility where blood cells are separated from the serum. These cells are lysed and we PCR amplify the proviral DNA, which is then subject to gel electrophoresis we send the product for sequencing analysis to identify SNPs, as well as functional analysis to quantify transcription activation.
add
So far we have a total of 91 brain samples: 15 normal, 46 minor cognitive motor disorder, 30 HADEarly trends show there is in fact a correlation between genetic variation with the LTR and neurocognitive statusa higher frequency of base pair changes in spleen derived tissue as compared to the brain 3T/4G compartmentalizes in specific brain regions such as Thalamus but and increases prevalence with disease severityOpp trend observed in 3T/6G which compartmentalizes with cerebellum but decreases in prevalence with disease severity