Ebolavirus Intervention 20181115

50 %
50 %
Information about Ebolavirus Intervention 20181115
Education

Published on November 16, 2018

Author: kylai23

Source: authorstream.com

Slide1: Management of Human Ebola Virus Infection Theoretical Pharmacological and Non-pharmacological Approach By Targeting The Host Response Dr. Lai Kang Yiu Queen Elizabeth Hospital, H.K.S.A.R. China BIT’s 9 th World Gene Convent Holiday Inn Singapore Atrium 13 – 15 Nov 2018 Slide2: 1997 Slide3: 31/7/14 Scientific Committee for Emerging and Zoonotic Diseases Chief Infection Control Officer Dr. Dominic Tsang Ngai-chong Slide4: We can have the solution only if we know what really happened ? Mattew 7:7 Ask, and it shall be given you; seek, and ye shall find; knock, and it shall be opened unto you: Slide5: The Extinction of Dinosaurs They have found a much better boat to house them! Slide6: WHO has advised that the use of experimental medications and vaccines under the exceptional circumstances of this outbreak is ethically justifiable. However, existing supplies of all these experimental medications are either extremely limited or exhausted . Before all these agents are available, in combating such unprecedented global public-health crisis, alternative intervention that might be effective against Ebola virus, are affordable, can be stockpiled and would be available on the first pandemic day should be explored to complement conventional therapy. Slide7: A Bee To Be Or A Fly To Be ? Ebola Virus Infection WHO Hong Kong Slide9: Revelation 3:8 Behold, I have set before you an open door, which no one is able to shut. “When one door closes, another door opens.” – Alexander Graham Bell, inventor of the telephone. Slide10: Uprising RNA Virus Epidemic and Pandemic Avian Influenza Viruses SARS Coronaviruses Ebola Virus MERS Coronavirus Slide11: 變幻原是永恆 The RNA polymerase of RNA virus has no proofreading mechanism. ~1 error per replicated genome  each cell can produce 10,000 new viral mutants to infect neighbouring cells. (300x than DNA virus) RNA Viruses Slide12: 月有陰晴圓缺 RNA Viruses SARS CoV MERS CoV Ebola We are only chasing the ever-changing shadow of the moon ! 如觀諸法生滅,悟世間無常。 Slide13: 窺破月影背後湛然不動的性 從月影的萬相 Unless we can understand the moon in a different perspective… Slide14: 月影背後 的永恆世界 The eternal world behind the shadow of the moon was formed 4.4-4.6 billions years ago Slide15: GP1,2 sGP ssGP Δ-peptide NP ( 核蛋白質 ) L-polymerase N 酸性蛋白 ) ( ( 一真法界 ,諸法平等,萬 相 皆殊。 Host VP30 VP40 VP35 VP24 Enveloped Filamentous non-segmented negative-sense, single- stranded RNA virus Slide16: HIV Virus Hepatitis C Virus A Combination Drug Regime May be needed to control these rapidly mutating RNA virus Why Cocktail Therapy: Why Cocktail Therapy Ebola viruses have underwent a rapid mutation ( 50 mutations within 1 month during its spread through humans). Ebola virus is RNA virus whose replication is highly error prone with nearly one viral mutation occurs during each cycle of replication. The genetic and antigenic diversity produced allows the viral population to evolve resistance to antiviral drugs and vaccines. Therefore combination therapy are introduced in the treatment of RNA virus infection such as HIV and hepatitis C virus , to prevent the develop of drug resistance. The goal of a cocktail regime containing anti-viral medications targeting different cycles of EBOV replication is to achieve maximal suppression of viral replication to prevent the rapid develop of EBOV to Favipiravir, the currently available medication that has been shown to reduce the replication of EBOV. A proposeed cocktail regime basing on available agent is in the next slide. http://www.sciencemag.org/content/early/2014/08/27/science.1259657.full.pdf http://www.nature.com/news/ebola-virus-mutating-rapidly-as-it-spreads-1.15777 http://www.nejm.org/doi/pdf/10.1056/NEJMoa1404505 Slide18: Being a RNA virus with relatively small coding capacity, Ebola viruses have taken advantage of the host cell functions for viral replication and intracellular trafficking of viral components and during the process has led to apoptosis of the host cells and both cytokine and interferon dysregulation Teicoplanin Slide19: Innate Immune Suppression Broad cell tropism Pathogenesis of Ebola Virus Acid pH Immune Dysregulation Vasomotor Dysregulation Plasmodium falciparum coinfection in 18% Mortality increase from 58% to 86% uncommon IL6/CRP ALT/AST CPK/Amylase/Urea/Cr Slide20: Bacterial co-infection is rare in patients with Ebola virus disease Lamb L, et al. J Infect. 2015 Sep;71(3):406-7. A negative EBOV RT-PCR 72 hours after symptom onset was required before discharge. The use of malaria RDTs NP and GI FilmArray can help to make alternative diagnosis within 72 hours. Diagnosis of Febrile Illnesses Other Than Ebola Virus Disease at an Ebola Treatment Unit in Sierra Leone. O'Shea MK, et al. Clin Infect Dis. 2015;61(5):795-8. Slide22: 10 9 to 10 10 virion/ml Slide24: Ebola hemorrhagic Fever: novel biomarker correlates of clinical outcome. McElroy AK et al. J Infect Dis. 2014;210(4):558-66. Slide25: JIKI trial mortality, according to age and baseline RT-PCR Ct value. Sissoko D, Laouenan C, Folkesson E, et al. Experimental Treatment with Favipiravir for Ebola Virus Disease (the JIKI Trial): A Historically Controlled, Single-Arm Proof-of-Concept Trial in Guinea. PLoS Med . 2016;13(3):e1001967. Published 2016 Mar 1. doi:10.1371/journal.pmed.1001967 EBOV viral load (log10 copies/ml) 6.8 (5.8 to 7.5) 8.7 (8.1 to 9.2) 8.8 (6.7 to 9.2) Slide26: Takada A, Kawaoka Y. The pathogenesis of Ebola hemorrhagic fever. Trends Microbiol. 2001 Oct;9(10):506-11. Slide27: Misasi J, Sullivan NJ. Camouflage and misdirection: the full-on assault of ebola virus disease. Cell . 2014;159(3):477-86. Slide28: Ebola virus: unravelling pathogenesis to combat a deadly disease. Hoenen T, Groseth A, Falzarano D, Feldmann H. Trends Mol Med. 2006;12:206-15. Slide29: Evade Antibody-associated innate immunity against in EBOV infection Mohan GS, Li W, Ye L, Compans RW, Yang C. Antigenic subversion: a novel mechanism of host immune evasion by Ebola virus. PLoS Pathog. Takada A, Ebihara H, Feldmann H, Geisbert TW, Kawaoka Y. Epitopes required for antibody-dependent enhancement of Ebola virus infection. J Infect Dis. 2007 Nov 15;196 Suppl 2:S347-56. Slide30: Two mechanisms for Antibody-dependent enhancement of Ebola virus infection Takada A, Watanabe S, Okazaki K, Kida H, Kawaoka Y. Infectivity-enhancing antibodies to Ebola virus glycoprotein. J Virol. 2001 Mar;75(5):2324-30. Slide31: EBOV GP Steric Shielding Younan P, Iampietro M, Bukreyev A. Disabling of lymphocyte immune response by Ebola virus. PLoS Pathog . 2018;14(4):e1006932. Published 2018 Apr 12. doi:10.1371/journal.ppat.1006932 Overexpression of EBOV GP on cell surface membrane impaired the development of T cell response Slide32: The catalytic domain of RNA-dependent RNA polymerase (RdRp) is conserved among various types of RNA viruses (arenaviruses, bunyaviruses and filoviruses Favipiravir-RTP (Favipiravir ribofuranosyl-5′-triphosphate) RdRp Influenza virus Furuta Y, et al. Favipiravir (T-705), a broad spectrum inhibitor of viral RNA polymerase. Proc Jpn Acad Ser B Phys Biol Sci . 2017;93(7):449-463. Slide33: RNA (-) strand Filoviridae Ebola 10.5 Low-dose D0 1000 mg favipiravir BID D1-D3 400 mg BID High-dose D0 1200 mg favipiravir BID D1-D3 800 mg BID Under phase 2 study in USA Slide34: 2014 23/3/2014 30/3/2014 10/5/2014 20/7/2014 11/8/2014 Funeral of a respected traditional healer 3956/14124 (28%) Sierra Leone–China Friendship hospital 10/10/2014 Favipiravir 1600mg D1 1200mg D2 3-11 day until discharge Liberian diplomat under observation got clearance and travelled by air to Nigeria for conference. 9 outbreak in 42 years Favipiravir D0 6000mg p.o. D1-D9 2400mg p.o.  well tolerated JIKI Trial Slide35: Guedj J, et al. Antiviral efficacy of favipiravir against Ebola virus: A translational study in cynomolgus macaques. PLoS Med . 2018;15(3):e1002535. Slide36: Host cell factors in filovirus entry: novel players, new insights. Hofmann-Winkler H et al. Viruses. 2012 Dec;4(12):3336-62. The clinically approved drugs amiodarone, dronedarone and verapamil inhibit filovirus cell entry. Gehring G et al. J Antimicrob Chemother. 2014 Aug;69(8):2123-31. Su-Yang Liu et al. 2012. Interferon-Inducible Cholesterol-25-Hydroxylase Broadly Inhibits Viral Entry by Production of 25-Hydroxycholesterol. Immunity , volume 38, issue 1, 92-105; pH-dependent (Chloroquine) Cat-L inhibitor (Teicoplanin) Amiodarone Clomiphene and toremifene Ebola virus entry requires the cholesterol transporter Niemann-Pick C1 IFN inducible Cholesterol-25-hydroxylase (CH25H), an enzyme that converts cholesterol to an oxysterol called 25-hydroxycholesterol (25HC), IFN- β IFITM (interferon-induced transmembrane) Antibodies β 1 integrins, DC-SIGNs, L-SIGNs, lectins, TIM-1s, Tyro3 family proteins, heparan sulfates, or folate receptor- α . TIM-1 Slide37: Diamond MS, Farzan M. The broad-spectrum antiviral functions of IFIT and IFITM proteins. Nat Rev Immunol . 2012;13(1):46-57 IFN- β IFITM3 SNP rs12252-C Slide38: Wrensch F, Karsten CB, Gnirss K, Hoffmann M, Lu K, Takada A, et al. Interferon-induced transmembrane protein-mediated inhibition of host cell entry of ebolaviruses. J Infect Dis (2015) 212(Suppl 2):S210–8. Slide39: Interferon- β therapy prolongs survival in rhesus macaque models of Ebola and Marburg hemorrhagic fever. Smith LM et al. J Infect Dis. 2013;208:310-8. 5 mg/kg monoclonal antibody to the IFNAR1 chain of the human type I IFN receptor administered 2 and 5 days after infection (n = 4) Recombinant human IFN-β 10.5 µg/kg, 18 hours and 1, 3 d, 5, 7, and 9 days after infection (n = 5; red line) Slide40: Filovirus entry: a novelty in the viral fusion world. Hunt CL et al. Viruses. 2012;4(2):258-75. &4 TIM-1&4 GP independent GP dependent Slide41: TAM TIM-1 (epithelial cells) (liver kidney adrenal gland and brain)  Cytokine dysregulation of T cell TIM-4 and TLR4 (macrophage and dendritic cells) Phosphatidylserine (PtdSer) receptors are for clearance of dying cells Phosphatidylserine receptors: enhancers of enveloped virus entry and infection. Moller-Tank S, Maury W. Virology. 2014 Nov;468-470:565-580. Slide42: Kuroda M, Fujikura D, Nanbo A, et al. Interaction between TIM-1 and NPC1 Is Important for Cellular Entry of Ebola Virus. J Virol . 2015;89(12):6481-93. Slide43: Ebola Viral Glycoprotein Bound to Its Endosomal Receptor Niemann-Pick C1. Wang H, Shi Y, Song J, Qi J, Lu G, Yan J, Gao GF. Cell. 2016 Jan 14;164(1-2):258-268. NPC1 pH 5-6 Slide44: Ebola Viral Glycoprotein Bound to Its Endosomal Receptor Niemann-Pick C1. Wang H, Shi Y, Song J, Qi J, Lu G, Yan J, Gao GF. Cell. 2016 Jan 14;164(1-2):258-268. Amiodarone: Amiodarone Block Ebola viral entry (99% reduction) at late endosome phase by inducing a Niemann-Pick C-like phenotype. Significant inhibition seen in most endothelial and epithelial cells ( macrophage, monocyte, endothelial cells ) except primary hepatocyte and fibroblast. IC 50 of amiodarone for EBOV is 0.25 ug/ml and serum level of amiodarone 1.5-2.5 ug/ml at dose for management of arrhythmia. Inhibitory effect on EBOV entry dose dependent and reversible upon removal of drug. Prolonged exposure to amiodarone will not lead to compensatory change in host cell. Available in both oral and intravenous preparation and both long term and short term side effect known. As a generic drug, its use would be feasible in both resource-rich and resource-poor setting. Slide46: Gehring G et al. J Antimicrob Chemother. 2014 Aug;69(8):2123-31. Serum level of amiodarone is 1.5-2.5 ug/ml at dose for management of arrhythmia Slide47: Amiodarone markedly reduces filoviral GP-mediated cell entry. at drug concentration 1.5 µg/mL and in 2.3 µM, respectively. Gehring G et al. J Antimicrob Chemother. 2014 Aug;69(8):2123-31. Slide48: Gehring G et al. J Antimicrob Chemother. 2014 Aug;69(8):2123-31. Filoviral GP-mediated entry into endothelial cells and in macrophages derived from primary human monocytes was significantly inhibited at amiodarone concentrations below 1 µg/mL while amiodarone had no effect on the transduction of primary hepatocytes and cell of fibroblast origin. Clomiphene and toremifene: Clomiphene and toremifene Clomiphene and toremifene does not disrupt the interaction between primed GP 1 and NPC1 but mediate the entry block indirectly through NPC1 by targeting other endosomal/lysosomal proteins involved in the cholesterol uptake pathway whose function may be regulated by NPC1. Clomiphene and toremifene at 60 mg/kg every other day have been shown to produce a 90% and 50% survival respectively in EBOV infected mice compared with 100% mortality in the control group in an in vivo murine Ebola infection model. They are effective in both male and female mice. Therapeutic dose against EBVO with tolerable side effect can be achieved with toremifene at an oral dose used in human trial for the treatment of advanced carcinoma of breast. However therapeutic dose against EBOV cannot be achieved with oral clomiphene. The peak concentration after oral admisinstration of toremifene is 4 hour. Toremifene undergoes extensive demethylation and hydroxylation to active and inactive metabolites via hepatic mixed function oxidases Hence amiodarone and toremifene may have a complementary role in protection against Ebola virus. Slide50: Johansen LM, Brannan JM, Delos SE, Shoemaker CJ, Stossel A, Lear C, Hoffstrom BG, Dewald LE, Schornberg KL, Scully C, Lehár J, Hensley LE, White JM, Olinger GG. FDA-approved selective estrogen receptor modulators inhibit Ebola virus infection. Sci Transl Med. 2013 Jun 19;5(190):190ra79. Clomiphene and toremifene at 60 mg/kg every other day have been shown to produce a 90% and 50% survival respectively in EBOV infected mice. Slide51: Both clomiphene and toremifene have anti-Ebola virus activity in both the Vero E6 (interferon-deficient African green monkey kidney epithelial cells) and HepG2 (human hepatocellular carcinoma) cell lines. Johansen LM wt al.FDA-approved selective estrogen receptor modulators inhibit Ebola virus infection. Sci Transl Med. 2013 Jun 19;5(190):190ra79. Slide52: Could estrogen-receptor antagonists treat Ebola? Josh Farkas http://www.pulmcrit.org/2014/08/could-estrogen-receptor-antagonists.html Slide53: Cat L Hood CL, Abraham J, Boyington JC, Leung K, Kwong PD, Nabel GJ. Biochemical and structural characterization of cathepsin L-processed Ebola virus glycoprotein: implications for viral entry and immunogenicity. J Virol . 2010;84(6):2972-82 Teicoplanin Chloroquine Slide54: Zhou N, Pan T, Zhang J, et al. Glycopeptide Antibiotics Potently Inhibit Cathepsin L in the Late Endosome/Lysosome and Block the Entry of Ebola Virus, Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV). J Biol Chem . 2016;291(17):9218-32. Trough concentration of Teicplanin at therapeutic dose 15 – 40 mg/L ( 8.78 μ m – 23.4 μ m) The IC 50 value of teicoplanin on the entry of Ebola trVLPs is as low as 330 nm, Slide55: After the completion of the loading dose regimens for most Gram-positive bacterial infections, the serum concentrations of teicoplanin are at least 15 mg/liter (8.78 μm), which is ∼27, 14, and 2 times higher than the IC50 values of teicoplanin against the entry of Ebola viruses, MERS-CoV/HIV-1, and SARS-CoV/HIV-1 pseudotyped viruses. Zhou N, Pan T, Zhang J, et al. Glycopeptide Antibiotics Potently Inhibit Cathepsin L in the Late Endosome/Lysosome and Block the Entry of Ebola Virus, Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV). J Biol Chem . 2016;291(17):9218-32. Influenza viruses SARS-CoV MERS-CoV Ebola HIV-1 HCV Flaviviruses FIPV Slide56: Zhou N, Pan T, Zhang J, et al. Glycopeptide Antibiotics Potently Inhibit Cathepsin L in the Late Endosome/Lysosome and Block the Entry of Ebola Virus, Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV). J Biol Chem . 2016;291(17):9218-32. Convalescent Plasma/Blood: Convalescent Plasma/Blood WHO issued a consensus statement that the use of whole blood therapies and convalescent blood serums needs to be considered as a matter of priority in the recent EBOV outbreak in West African countries. The development of neutralizing antibody and T cell response are important for recovery from Ebola virus infection. Patients who are able to mount an immune response to the virus will begin to recover in 7 to 10 days and start a period of prolonged convalescence. In survivors, early and increasing levels of IgG, directed mainly against the nucleoprotein and the VP40, were followed by clearance of circulating viral antigen and activation of cytotoxic T cells. Convalescent blood has been shown to improve survival of EBOV infected patient during the outbreak of EBOV in Kikwit in 1995. Investigations of antibodies against Ebola virus showed that non-neutralizing antibodies recognized a GP epitope in the non-essential mucin-like domain while neutralizing antibody were specific for GP1. In addition, neutralizing antibodies from survivors has impaired recognition for sGP and binding was dependent on presence of GP2 residues which are not present in sGP. These studies have laid the foundation for subsequent clinical research on the use of monoclonal antibodies such as Zmapp in the treatment of Ebola virus infection. Slide59: 4/14 has neutralizing antibodies (4/4 GP 2/4 VP40) Since first identified almost 40 years ago, the genome of the Ebola has shown remarkable stability, an unusual features for RNA virus with an error prone polymerase. Misasi J, Sullivan NJ. Camouflage and misdirection: the full-on assault of ebola virus disease. Cell . 2014;159(3):477-86. GP (12/14) NP (13/14) VP40 (3/14) Slide60: Cook JD, Lee JE. The secret life of viral entry glycoproteins: moonlighting in immune evasion. PLoS Pathog. 2013;9(5):e1003258. West BR, Moyer CL, King LB, et al. Structural Basis of Pan-Ebolavirus Neutralization by a Human Antibody against a Conserved, yet Cryptic Epitope. MBio . 2018;9(5):e01674-18. Published 2018 Sep 11. doi:10.1128/mBio.01674-18 ADI-15878 ADI-15878 targeting GP internal fusion loop (IFL) is able to neutralize all 5 strains of Ebola virus “ Base-binding” and fusion loop-binding epitope antibodies HR1 & HR2 regions RBS Internal fusion loop The Tetherin Antagonism of the Ebola Virus Glycoprotein Requires an Intact Receptor-Binding Domain and Can Be Blocked by GP1-Specific Antibodies. Slide61: Murin CD, Bruhn JF, Bornholdt ZA, Copps J, Stanfield R, Ward AB. Structural Basis of Pan-Ebolavirus Neutralization by an Antibody Targeting the Glycoprotein Fusion Loop. Cell Rep . 2018;24(10):2723-2732.e4. The HR1 and IFL on filoviral GPs are highly conserved in sequence and structure across genera ADI-15878 targeting GP internal fusion loop is able to neutralize all 5 strains of Ebola virus Slide62: In the ebolaviruses, the IFL is tucked beneath the cathepsin cleavage loop, which is loosely tucked in between GP protomers. Murin CD, Bruhn JF, Bornholdt ZA, Copps J, Stanfield R, Ward AB. Structural Basis of Pan-Ebolavirus Neutralization by an Antibody Targeting the Glycoprotein Fusion Loop. Cell Rep . 2018;24(10):2723-2732.e4. Internal fusion loop Slide63: West BR, Moyer CL, King LB, et al. Structural Basis of Pan-Ebolavirus Neutralization by a Human Antibody against a Conserved, yet Cryptic Epitope. MBio . 2018;9(5):e01674-18. Published 2018 Sep 11. doi:10.1128/mBio.01674-18 KZ52 and other base-binding antibodies lock down the GP2 N terminus and prevent it from untethering. In contrast, ADI-15878 uses a strategy to bind underneath the GP2 N-terminal tail and replace the hydrophobic tail-pocket interactions with its own hydrophobic antibody-pocket interactions, potentially inhibiting any as yet undiscovered downstream steps in the GP fusion pathway. Slide64: Phoolcharoen W1, Bhoo SH, Lai H, Ma J, Arntzen CJ, Chen Q, Mason HS.Expression of an immunogenic Ebola immune complex in Nicotiana benthamiana. Plant Biotechnol J. 2011 Sep;9(7):807-16 Slide65: Phoolcharoen W1, Bhoo SH, Lai H, Ma J, Arntzen CJ, Chen Q, Mason HS.Expression of an immunogenic Ebola immune complex in Nicotiana benthamiana. Plant Biotechnol J. 2011 Sep;9(7):807-16 Icon Genetics Provides Technology for Possible Ebola Treatment Indicator proteins glow under ultraviolet light on the leaves of the nicotiana benthamiana plant, which is a close relative of tobacco. Nicotiana benthamiana is used in the production of antibodies for Zmapp Slide66: Cytokine dysregulation and apoptosis of lympocytes Slide67: HA /GP mucin domain Activation of NF- κ B is caused by the accumulation of proteins in the endoplasmic recticulum membrane (ER) , a condition called ER-overload. Both the release of Ca2+ from the ER and the subsequent production of reactive oxygen intermediates are required for ER-overload-mediated NF- κ B activation. The ER-overload response: activation of NF-kappa B. Pahl HL. Baeuerle PA. Trends in Biochemical Sciences. 22(2):63-7, 1997 Feb. Signal Transduction From the Endoplasmic Reticulum to the Cell Nucleus Heike L. Pahl Physiological Reviews, Vol. 79, No. 3, July 1999, pp. 683-701 Role of NAC / Vit C Slide68: Cytokine Storm TNF-alpha, IL-1beta, IL-6, and IL-8 . Expression of influenza virus hemagglutinin activates transcription factor NF-kappa B HL Pahl and PA Baeuerle J. Virol., Mar 1995, 1480-1484, Vol 69, No. 3 The cytokines storm of EBOV is through the activation of NF-ķB by reactive oxygen species Slide69: 傳播 繁殖 Ebola virus selectively inhibits responses to interferons, but not to interleukin-1beta, in endothelial cells. Harcourt BH et al. J Virol. 1999 Apr;73(4):3491-6. Ebola virus Cytopathic damage and cytokine dysregulation lead to necrosis of cells to facilitate nuclear export of vRNP Replication Propagation Slide70: Ebola Virus ER-overload induces ROS Cytosol IRF-3 NF κβ Anti-viral activity (IFN α / β ) Anti-oxidants block ROS Pro-inflammatory activity IFN α / β , TNF- α & IL-1 β hTIM-1 GP mucin domain NF κ B-induced Cytokine Dysregulation of Ebola Virus infection Ibuprofen Block NF κ B activation EBOV and Shed GPs Remove by haemofilter TLR4 PtdSer Slide71: Pleet ML, DeMarino C, Lepene B, Aman MJ, Kashanchi F. The Role of Exosomal VP40 in Ebola Virus Disease. DNA Cell Biol. 2017;36(4):243-248. T cell Monocyte Slide72: Pleet ML, DeMarino C, Lepene B, Aman MJ, Kashanchi F. The Role of Exosomal VP40 in Ebola Virus Disease. DNA Cell Biol. 2017;36(4):243-248. T cell Monocyte VP40 within the Ebola-infected host cell will interact with the ESCRT pathway and be packaged into exosomes. Exosomes containing EBOV VP40 exit the cell and downregulate miRNA machinery (i.e., Dicer, Drosha, Ago 1) in recipient cells, including T-cells and monocytes/macrophages (MØ). Subsequent bystander lymphocyte apoptosis is increased, potentially resulting in immune system repression and increased viral replication in various compartments of the host.: VP40 within the Ebola-infected host cell will interact with the ESCRT pathway and be packaged into exosomes. Exosomes containing EBOV VP40 exit the cell and downregulate miRNA machinery (i.e., Dicer, Drosha, Ago 1) in recipient cells, including T-cells and monocytes/macrophages (MØ). Subsequent bystander lymphocyte apoptosis is increased, potentially resulting in immune system repression and increased viral replication in various compartments of the host. Oxytetracycline Ebola VP40 in Exosomes Can Cause Immune Cell Dysfunction. Pleet ML et al. Front Microbiol. 2016;7:1765. Model for Ebola VP40 role in pathogenesis Slide74: Escudero-Pérez B, Volchkova VA, Dolnik O, Lawrence P, Volchkov VE. Shed GP of Ebola virus triggers immune activation and increased vascular permeability. PLoS Pathog. 2014 Nov 20;10(11):e1004509. Exosome VP40 Slide76: Salvador B, Sexton NR, Carrion R Jr, Nunneley J, Patterson JL, Steffen I, Lu K, Muench MO, Lembo D, Simmons G. Filoviruses utilize glycosaminoglycans for their attachment to target cells. J Virol. 2013 Mar;87(6):3295-304. A rapid device-based therapy for treatment of Ebola and Marburg infections.http://extheramedical.com/images/EbolaandMarburg.pdf Filoviruses utilize glycosaminoglycans for their attachment to target cells. Ebola and Marburg virus have been shown to bind to both surface-bound (immobilized) and soluble heparin Physical Removal of Ebola virus To Reduce Viral Load ! Slide77: Aethlon ADAPT (adaptive dialysis-like affinity platform technology) System https://rglmd.wordpress.com/ Reduce viral load and cytokine dysregulation Aethlon’s Hemopurifier device: Aethlon’s Hemopurifier device The cartridge is filled with about 2,800 hollow fibers that blood flows through, Joyce said. The fibers are porous to particles smaller than 250 nanometers. Ebola virus particles are 80 nanometers wide , so they pass through the fiber into the space between the cartridge shell and the fibers. Blood cells are too large to pass through, so they remain inside the fibers. Outside the fibers, proteins called lectins are attached to the cartridge. These particular lectins have an affinity for viral glycoproteins, a complex of proteins and sugars, especially one called mannose. The viruses stick to the lectins, while fluids and other particles return to the fibers and ultimately back into the patient’s bloodstream. Viral glycoprotein fragments which induce immune evasion and cytokine dysregulation also are trapped by the Hemopurifier. Aethlon ADAPT (adaptive dialysis-like affinity platform technology) system: Aethlon ADAPT (adaptive dialysis-like affinity platform technology) system Bind exosomes (30-100nm), glycoproteins residing on the envelope of viruses and viral particles . As the patient’s blood passes through the device, plasma component < 200 nm is size travel through the porous fibers and interact with the immobilized affinity agents Dr. Helmut Geiger, the chief of nephrology at Frankfurt University Hospital in Germany has presented his finding at the nephrology society’s conference in Philadelphia. Before treatment with Aethlon’s Hemopurifier, the patient had 400,000 virus particles per milliliter of blood and the “viral load” dropped to 1,000 per milliliter after the dialysis (6.5hours) . There was no further rise above that level again. To prove the Hemopurifier actually removed Ebola viruses from Mawanda’s bloodstream, the device was flushed after the treatment course. Researchers at Philipps University of Marburg — in Marburg, Germany, then measured the virus particles they found. Those researchers said 242 million Ebola viruses were captured by the Hemopurifier. In general, ebolavirions are 80 nanometers (nm) in width and may be as long as 14,000 nm. Slide80: Severe Ebola virus disease with vascular leakage and multiorgan failure: treatment of a patient in intensive care. Wolf T, et al. Lancet. 2015;1428-35. Slide81: Severe Ebola virus disease with vascular leakage and multiorgan failure: treatment of a patient in intensive care. Wolf T, et al. Lancet. 2015;1428-35. RRT Slide82: Management of Vascular Dysregulation of Ebola virus Infection Slide83: Interleukin-6 regulates the zinc transporter Zip14 in liver and contributes to the hypozincemia of the acute-phase response. Liuzzi JP et al. Proc Natl Acad Sci U S A. 2005:6843-8. Slide84: Kimura T; Kambe T. International Journal of Molecular Sciences. 17(3):336, 2016 50% 30-40% 10% Zrt- and Irt-like proteins Zn transporters Metallothioneins (MTs) Slide85: Augusto C. Montezano et al. ROS and endothelial cell function Basic & Clinical Pharmacology & Toxicology, 110, 87–94 Slide86: Mechanisms of group A Streptococcus resistance to reactive oxygen species. Henningham A, Döhrmann S, Nizet V, Cole JN. FEMS Microbiol Rev. 2015;39:488-508. Slide88: Mechanism of action of vitamin C in sepsis: ascorbate modulates redox signaling in endothelium. Biofactors . 2009;35(1):5-13. Slide89: Ebola virus modulates transforming growth factor β signaling and cellular markers of mesenchyme-like transition in hepatocytes. Kindrachuk J et al. J Virol. 2014 ;88(17):9877-92. EBOV infection modulated TGF- β -mediated signaling responses and promoted “mesenchyme-like” phenotypic changes Inhibition of TGF- β production inhibited viral replication in vitro and reduced EBOV pathogenesis in vivo. Slide90: Pentoxifylline (NFkB/TNF- α inhibitor) Transforming growth factor-beta (TGF- β ) Inhibit both fibroblast proliferation Inhibit collagen synthesis Zinc Enhanced re-epithelialization of respiratory epithelium Selenium Glutathione peroxidase (GPx) is Selenium dependent Glutathione reductase (GR) require riboflavin (FAD) GPx GR Prevent peroxidative damage Slide91: Transforming growth factor-beta (TGF- β ) Endothelial-mesenchymal transdifferentiation Epithelial-mesenchymal transdifferentiation Slide92: Alveolar epithelial disintegrity in pulmonary fibrosis. Kulkarni T, et al. Am J Physiol Lung Cell Mol Physiol. 2016:L185-91. Structural disruption of TJs with subsequent loss of alveolar epithelial integrity play an important role in the development of pulmonary fibrosis. Slide93: Alveolar epithelial disintegrity in pulmonary fibrosis. Kulkarni T, et al. Am J Physiol Lung Cell Mol Physiol. 2016:L185-91. TGF-β1 causes disruption of TJs in human alveolar epithelial cells and induce epithelial-to-mesenchymal transition (EMT). TJ: Barrier function and alveolar fluid homeostasis NF κ B-induced TNF- α reduces claudin essesntial for restoring alveolar epithelial barrier function. TNF- α Slide94: vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 (Flk-1) SARS-CoV nucleocapsid (N) protein inhibits apoptosis in favor of virus packaging and replication at the early stage of SARS development. N protein potentiates TGF- β 1 induced PAI-1 expression leading to development of lung fibrosis at the late stage . TGF- β downregulate the expression of extracellular matrix proteases and stimulate the expression of extracellular matrix protease inhibitors. PAI-1, which is the primary inhibitor of both tissue-type and urokinase-type plasminogen activator is a well established target of TGF- β via the Smad pathway and plays a pivotal role in TGF- β -promoted tissue fibrosis. The mean concentration of serum TGF- β 1 in SARS patients was higher than that of the control group during all clinical courses. Although TGF- β 1 in serum decreased in remission and recovery stage in SARS patients, the average was still higher than that of the control group (P < 0.01). Treating the Host Response to Ebola Virus Disease with Generic Statins and Angiotensin Receptor Blockers ? 98% survival rate : Treating the Host Response to Ebola Virus Disease with Generic Statins and Angiotensin Receptor Blockers ? 98% survival rate Around 100 patients with laboratory-confirmed Ebola virus disease at the 34 Military Hospital in Freetown, the Port Loko Government Hospital, the Hastings Ebola Treatment Centre, and other sites in Sierra Leone. Patients were given atorvastatin (40 mg/day) and irbesartan (150 mg/day). Reports indicate that rapid clinical improvement was seen in almost all patients, and only two who were inadequately treated are known to have died (O.M.R.,unpublishedobservations).One was critically ill when first seen and died soon thereafter. The other initially responded to 3 days of combination treatment, but treatment was stopped and he was given an antiviral agent, he relapsed and died. Fedson DS, Jacobson JR, Rordam OM, Opal SM. Treating the Host Response to Ebola Virus Disease with Generic Statins and Angiotensin Receptor Blockers. MBio . 2015;6(3):e00716. Slide98: Quarantine (Hippocrates) ( 希波克拉底 ) Leviticus 13/4-5.  The priest is able to isolate person with suspected leprosy for 7 + 7 days Venice :1348 A council of three the power to detain ships, cargoes, and individuals in the Venetian lagoon for up to 40 days . Mass Plague Graves on Venice "Quarantine" Island Lazzaretto Nuovo 1377 Law of Ragusa “Trentina” and “Quaranta” ) 1423 Venice 1 st “Lazaretto” (Quarantine station) Rat guard Rat Guard Slide99: Isolation And Quarantine Policy Slide100: Thank you for your attention References: References Sissoko D, et al. Experimental Treatment with Favipiravir for Ebola Virus Disease (the JIKI Trial): A Historically Controlled, Single-Arm Proof-of-Concept Trial in Guinea. PLoS Med . 2016;13(3):e1001967. Furuta Y, Komeno T, Nakamura T. Favipiravir (T-705), a broad spectrum inhibitor of viral RNA polymerase. Proc Jpn Acad Ser B Phys Biol Sci . 2017;93(7):449-463. Zhou N, Pan T, Zhang J, et al. Glycopeptide Antibiotics Potently Inhibit Cathepsin L in the Late Endosome/Lysosome and Block the Entry of Ebola Virus, Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV). J Biol Chem . 2016;291(17):9218-32. Hood CL, Abraham J, Boyington JC, Leung K, Kwong PD, Nabel GJ. Biochemical and structural characterization of cathepsin L-processed Ebola virus glycoprotein: implications for viral entry and immunogenicity. J Virol . 2010;84(6):2972-82. References: References Mechanisms of Filovirus Entry. Davey RA, Shtanko O, Anantpadma M, Sakurai Y, Chandran K, Maury W. Curr Top Microbiol Immunol. 2017;411:323-352. Filovirus entry: a novelty in the viral fusion world. Hunt CL, Lennemann NJ, Maury W. Viruses. 2012 Feb;4(2):258-75. Structural Basis of Pan-Ebolavirus Neutralization by an Antibody Targeting the Glycoprotein Fusion Loop. Murin CD, Bruhn JF, Bornholdt ZA, Copps J, Stanfield R, Ward AB. Cell Rep. 2018 Sep 4;24(10):2723-2732.e4. doi: 10.1016/j.celrep.2018.08.009. Characterization of Human and Murine T-Cell Immunoglobulin Mucin Domain 4 (TIM-4) IgV Domain Residues Critical for Ebola Virus Entry. Rhein BA, Brouillette RB, Schaack GA, Chiorini JA, Maury W. J Virol. 2016 Jun 10;90(13):6097-6111. Phosphatidylserine receptors: enhancers of enveloped virus entry and infection. Moller-Tank S, Maury W. Virology. 2014 Nov;468-470:565-580. Bornholdt ZA, Ndungo E, Fusco ML, et al. Host-Primed Ebola Virus GP Exposes a Hydrophobic NPC1 Receptor-Binding Pocket, Revealing a Target for Broadly Neutralizing Antibodies. MBio . 2016;7(1):e02154-15. Published 2016 Feb 23. doi:10.1128/mBio.02154-15 West BR, Moyer CL, King LB, et al. Structural Basis of Pan-Ebolavirus Neutralization by a Human Antibody against a Conserved, yet Cryptic Epitope. MBio . 2018;9(5):e01674-18. Published 2018 Sep 11. doi:10.1128/mBio.01674-18 Murin CD, Bruhn JF, Bornholdt ZA, Copps J, Stanfield R, Ward AB. Structural Basis of Pan-Ebolavirus Neutralization by an Antibody Targeting the Glycoprotein Fusion Loop. Cell Rep . 2018;24(10):2723-2732.e4. Keck ZY, Enterlein SG, Howell KA, et al. Macaque Monoclonal Antibodies Targeting Novel Conserved Epitopes within Filovirus Glycoprotein. J Virol . 2015;90(1):279-91. Published 2015 Dec 17. doi:10.1128/JVI.02172-15 References: References Holtsberg FW, Shulenin S, Vu H, et al. Pan-ebolavirus and Pan-filovirus Mouse Monoclonal Antibodies: Protection against Ebola and Sudan Viruses. J Virol . 2015;90(1):266-78. Published 2015 Dec 17. doi:10.1128/JVI.02171-15 Rimoin AW, Lu K, Bramble MS, et al. Ebola Virus Neutralizing Antibodies Detectable in Survivors of theYambuku, Zaire Outbreak 40 Years after Infection. J Infect Dis . 2017;217(2):223-231. Khanna M, Sharma S, Kumar B, Rajput R. Protective immunity based on the conserved hemagglutinin stalk domain and its prospects for universal influenza vaccine development. Biomed Res Int . 2014;2014:546274. . Kirkpatrick E, Qiu X, Wilson PC, Bahl J, Krammer F. The influenza virus hemagglutinin head evolves faster than the stalk domain. Sci Rep . 2018;8(1):10432. Published 2018 Jul 11. doi:10.1038/s41598-018-28706-1 Takada A, Feldmann H, Ksiazek TG, Kawaoka Y. Antibody-dependent enhancement of Ebola virus infection. J Virol . 2003;77(13):7539-44. Takada A, Watanabe S, Okazaki K, Kida H, Kawaoka Y. Infectivity-enhancing antibodies to Ebola virus glycoprotein. J Virol . 2001;75(5):2324-30. Younan P, Iampietro M, Bukreyev A. Disabling of lymphocyte immune response by Ebola virus. PLoS Pathog . 2018;14(4):e1006932. Published 2018 Apr 12. doi:10.1371/journal.ppat.1006932 References: References McElroy AK, Erickson BR, Flietstra TD, et al. Von Willebrand factor is elevated in individuals infected with Sudan virus and is associated with adverse clinical outcomes. Viral Immunol . 2015;28(1):71-3. The Ebola virus soluble glycoprotein (sGP) does not affect lymphocyte apoptosis and adhesion to activated endothelium. Wolf K, et al. J Infect Dis. 2011:S947-52. Misasi J, Sullivan NJ. Camouflage and misdirection: the full-on assault of ebola virus disease. Cell . 2014;159(3):477-86. Mechanism of action of vitamin C in sepsis: ascorbate modulates redox signaling in endothelium. Biofactors . 2009;35(1):5-13. Fedson DS, Jacobson JR, Rordam OM, Opal SM. Treating the Host Response to Ebola Virus Disease with Generic Statins and Angiotensin Receptor Blockers. MBio . 2015;6(3):e00716. Published 2015 Jun 23. doi:10.1128/mBio.00716-15 Generic Statins and Angiotensin Receptor Blockers: Are They Really Useful in Ebola? Viroj Wiwanitkit mBio. 2016 Jan-Feb; 7(1): e02228-15.

Add a comment

Related presentations