Evening Ebola Update, Tues, 12/10: BSL-4 lab in Boston not open for business/Genomics in I.D. in Africa/ More on hemopurifiers   Leave a comment

12/10/14

Dear Colleagues:

l.  Boston.com, a Boston on-line magazine, has an article ‘Why Haven’t We Found A Cure for EBOV in Boston?’.  The article notes that there are several researchers in Boston (Sabeti, Conner, Muhlberger to name a few) who have done/are doing exciting research to stop the EBOV epidemic in West Africa.  Yet a Biosafety Level 4 (BSL-4) lab at Boston University is still not open for EBOV researchAll EBOV research must be done in a BSL-4 lab.  The article says that political, regulatory, and financial reasons lie behind the delay in opening this necessary BSL-4 lab.  See the article at: http://www.boston.com/health/2014/12/08/why-haven-found-cure-for-ebola-boston/MSlcjIbkLmuNCycszqhJGJ/story.html. 

2. Dr. Racaniello actually went into the Boston BSL-4 lab-to-be.  His netcast can be viewed at: http://www.twiv.tv/threading-the-neidl/

3.  Genome Biology on 7 November contains an article by Folarin, et. al. from Redeemer’s University in Nigeria which tells us about African plans to establish a genomic center for infectious disease control within Africa.  The effort will be backed by NIH, Wellcome Trust, and the World Bank.  See details of the plans at: http://genomebiology.com/2014/15/11/515

4.  CDC’s MMWR 9 December has a report on the incidence of EBOV in HCW in Sierra Leone from May through October, 2014.  Five percent of HCW contracted EBOV during this time period.  Contributing factors included: inadequate PPEs, poor triage of suspected EBOV patients, lack of ambulances, time delay between blood samples for EBOV testing and return of test results, no delineation of low risk and high risk EBOV wards, lack of control of EBOV patient movement within treatment center, no HCW training for EBOV patients, inadequate hand washing.  See more details in the MMWR at: http://www.cdc.gov/mmwr/preview/mmwrhtml/mm63e1209a2.htm?s_cid=mm63e1209a2_e

5.  Based on the rate of EBOV in HCW in Sierra Leone at the end of October, the CDC dispatched an Ebola Response Team to Sierra Leone to investigate causes of the HCW infections and provide instruction to the Ministry of Health.  The team’s findings and recommendations to the Ministry of Health are reported in the CDC MMWR 9 December by Pathmanathan, et. al.  See the report at: http://www.cdc.gov/mmwr/preview/mmwrhtml/mm63e1209a3.htm?s_cid=mm63e1209a3_e

6.  Secretary Sylvia Burwell of Health and Human Services in the U.S. issued today a memorandum giving legal immunity in the United States to the vaccine makers, distributors, and testers of the GSK, NewLink, and Johnson & Johnson vaccines soon to be tested in West Africa.  Her announcement corresponds with a WHO meeting on 12 December of the involved parties.  This one-day meeting is to determine how to expedite the testing in West Africa.  See Secretary Burwell’s document at: http://www.hhs.gov/news/press/2014pres/12/20141209a.html.  Note the legal immunity does not apply to claims in courts outside the U.S.

7.  ACAPS lists Severe Humanitarian Crisis in Sierra Leone because 537 cases of EBOV reported the week ending November 30th and DRC because 50+ camps for internally displaced persons (IDP) were closed because the government said weapons were found in these camps.  See details at: http://geo.acaps.org/

8.  KY Lai of the Hospital Authority of Hong Kong has provided us with more information on the history of devices to filter viruses from the blood, includin the Aethlon Hemopurifier discussed in a previous posting.  His comments follow:

BEGIN COMMENTS:

The use of extracorporeal hemo-sorbent and hemodialysis for the treatment of human Marburg virus infection due to laboratory exposure in Russia was first reported in 1994.[1] Despite positive outcome, the actual value of this treatment approach is questioned.[2] Because of the concern on bioterrorism after 911,[3],[4] the physical removal of virus and soluble viral glycoproteins based on affinity adsorption was resurrected as part of the broad-spectrum treatment countermeasure against category “A” bioterror threats.[5],[6] Many bacteria and viruses depends on cell-surface heparan sulfate for their attachment to host cell.[7],[8] Ebola and Marburg virus have been shown to bind to both surface-bound (immobilized) and soluble heparin.[9] Hence haemofiltration filter which has heparin covalently coated to the filter membrane surface has been advocated as an adsorption media to remove intravenous Ebola virus particles and their soluble glycoproteins.[10] More targeted therapy using anti-viral antibody coated membrane filter has been studied in the treatment of HIV infection.[11],[12] The viral load, histologic abnormalities and liver damages of HCV infection is observed to be milder in hemodialysis patients despite the immune compromise caused by chronic uremia. A severe clinical course of chronic hepatitis C is unusual in most hemodialysis (HD) patients.[13],[14] This was attributed to a reduction in HCV viral load during each HD session.[15],[16] A cocktail drug therapy is needed in the control of human infection with chronic hepatitis C virus, an encapsulated RNA virus similar to Ebola virus. HCV viral load reduction via haemofiltration therapy has the advantage of reducing viral load without adding the risk of drug toxicity and drug interaction. This may be of advantageous in patient not tolerating multi-medications or for the management of multi-drug resistant strains. Hepatitis C is around 50 nm in size[17] and hence can be filtered out by most high flux and high efficiency haemofilter filter.

 

Continuous venovenous hemodialysis using high flux, high efficiency haemofilter and regional citrate anticoagulation has been employed in Emory University Hospital for a patient with Ebola virus infection complicated by acute renal failure. Despite the detection of virus inside the urine and blood, all three samples of CRRT efflux tested by aRT-PCR has no detectable viral particles indicating that there is no evidence of transfer of genetic material across the CRRT membrane to the spent effluent. Ebola virus is 80nm in diameter and 800 to 1000 nm long with a molecular size of 4200 kD. The high flux and high efficiency high flux filters used are impermeable to substance above a molecular mass of 60-70kD. No pre-filter and post-filter viral load was tested in the study.[18] Viral load reduction has been demonstrated by the use of an Aethlon Hemopurifier in one Ebola infected patient, Michael Mawanda, in Germany.[19] Aethlon Hemopurifier is a selective filtration device containing affinity agents that tightly bind to high-mannose structures unique to the surface of exosomes (30-100nm) produced by cancer and glycoproteins residing on the envelope of viruses. While most dialysis filters can pull out molecules that are less than 4 nanometers in diameter, this filter allow rapid extracorporeal capture and selective retention of target particles less than 200 nanometers from the entire circulatory system. This Aethlon ADAPT (adaptive dialysis-like affinity platform technology) system consists of immobilized affinity agents in the outer capillary space of hollow-fiber plasma separator cartridges that integrate into standard dialysis unit or continuous renal replacement therapy (CRRT) machines. 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 to which target molecules are selectively adsorbed while blood cells and non-bound serum components pass through the device. The matrices can be customized with one or two more affinity substances comprising monoclonal antibodies, lecithins, aptamers or other affinity agents to specificially capture and remove tumour-derived exosomes and other soluble viral glycoproteins and viral particles.[20] 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. 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. Doctor Geiger. Michael Connor Jr. and Harold Franch, who have treated Ebola patients in Emory University, commented that survival can’t be attributed to the Hemopurifier as the patient has also received other therapy.

 

This may be another alternative measures for the management of the profound viraemia and cytokine dysregulation in advanced EBOV infection with high viral load. The cytokine dysregulation of EBOV is due to both ROS-dependent NFkB activation of EBOV infected cells[21],[22],[23],[24],[25],[26] and the TLR-4 dependent activation of uninfected cells by the shed GP1-GP2 particles.[27] The molecular mass of shed GP is ~160 kDa and 7-10 nm long. High dose N-acetylcysteine can probably suppress the ROS-dependent NFkB activation of EBOV infected cells but probably unable to control the TLR4-dependent cytokine dysregulation of the uninfected cell induced by the shed GP1-GP2. These shed GP1-GP2 protein can be effectively removed by the Aethlon’s Hemopurifier. This may be important when the Ebola virus become resistant to the target therapy under development due to rapid viral mutation during an EBOV pandemic. In this worst case scenario, treatment targeting host cell factors and Aethlon’s Hemopurifier may have a complementary role as these are independent of EBOV mutation.

END COMMENTS.

REFERENCES:

 1] Nikiforov VV, Turovskii Iu I, Kalinin PP, et al. [A case of a laboratory infection with Marburg fever] Zh. Mikrobiol. Epidemiol. Immunobiol. 1994;(3):104–106.

[2] Mehedi M, Groseth A, Feldmann H, Ebihara H. Clinical aspects of Marburg hemorrhagic fever. Future Virol. 2011 Sep;6(9):1091-1106.

[3] Bray M. Defense against filoviruses used as biological weapons. Antiviral Res. 2003 Jan;57(1-2):53-60.

[4] Peters CJ. Marburg and Ebola–arming ourselves against the deadly filoviruses. N Engl J Med. 2005 Jun 23;352(25):2571-3.

[5] A Platform Technology to Address Bioterrorism http://www.aethlonmedical.com/assets/001/5040.pdf

[6] Ambrus JL Sr, Ambrus JL Jr, Ambrus CM. Therapy of newly emerging mutant viral disorders and role in bioterrorism. Med Hypotheses. 2005;64(6):1248-9.

[8] Rostand KS, Esko JD. Microbial adherence to and invasion through proteoglycans. Infect Immun. 1997 Jan;65(1):1-8.

[9] 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.

[10] A rapid device-based therapy for treatment of Ebola and Marburg infections. http://extheramedical.com/images/EbolaandMarburg.pdf

[11] Tullis RH, Duffin RP, Zech M, Ambrus JL. Affinity hemodialysis for antiviral therapy. II. Removal of HIV-1 viral proteins from cell culture supernatants and whole blood. Blood Purif. 2003;21(1):58-63.

[12] Tullis RH, Duffin RP, Zech M, Ambrus JL Jr. Affinity hemodialysis for antiviral therapy. I. Removal of HIV-1 from cell culture supernatants, plasma, and blood. Ther Apher. 2002 Jun;6(3):213-20.

[13] Fabrizi F, Messa P, Martin P. Impact of hemodialysis therapy on hepatitis C virus infection: a deeper insight. Int J Artif Organs. 2009 Jan;32(1):1-11.

[14] Fabrizi F, Dixit V, Messa P, Martin P. Hepatitis C-related liver disease in dialysis patients. Contrib Nephrol. 2012;176:42-53.

[15] Tullis RH, Duffin RP, Handley HH, Sodhi P, Menon J, Joyce JA, Kher V. Reduction of hepatitis C virus using lectin affinity plasmapheresis in dialysis patients. Blood Purif. 2009;27(1):64-9.

[16] Fabrizi F, Bunnapradist S, Lunghi G, Martin P. Kinetics of hepatitis C virus load during hemodialysis: novel perspectives. J Nephrol. 2003 Jul-Aug;16(4):467-75.

[18] Connor MJ Jr, Kraft C, Mehta AK, Varkey JB, Lyon GM, Crozier I, Ströher U, Ribner BS, Franch HA. Successful Delivery of RRT in Ebola Virus Disease. J Am Soc Nephrol. 2014 Nov 14.

[20] Marleau AM, Chen CS, Joyce JA, Tullis RH. Exosome removal as a therapeutic adjuvant in cancer. J Transl Med. 2012 Jun 27;10:134.

[21] Wahl-Jensen V, Kurz SK, Hazelton PR, Schnittler HJ, Ströher U, Burton DR, Feldmann H: Role of Ebola virus secreted glycoproteins and virus-like particles in activation of human macrophages. J Virol 2005, 79:2413–2419.

[22] Yang ZY, Duckers HJ, Sullivan NJ, Sanchez A, Nabel EG, Nabel GJ: Identification of the Ebola virus glycoprotein as the main viral determinant of vascular cell cytotoxicity and injury. Nat Med 2000, 6:886–889.

[23] Martinez O, Valmas C, Basler CF: Ebola virus-like particle-induced activation of NF-kappaB and Erk signaling in human dendritic cells requires the glycoprotein mucin domain. Virology 2007, 364:342–354.

[24] Volchkov VE, Volchkova VA, Muhlberger E, Kolesnikova LV, Weik M, Dolnik O, Klenk HD: Recovery of infectious Ebola virus from complementary DNA: RNA editing of the GP gene and viral cytotoxicity. Science 2001, 291:1965–1969.

[25] Pahl HL, Baeuerle PA: The ER-overload response: activation of NF-kappa B. Trends Biochem Sci 1997, 22:63–67.

[26] Pahl HL, Baeuerle PA: Activation of NF-kappa B by ER stress requires both Ca2+ and reactive oxygen intermediates as messengers. FEBS Lett 1996, 392:129–136.

[27] 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, 10:e1004509.

My thanks to Dr. Lai for his contribution to this blog.

RGL, MD

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Posted December 10, 2014 by levittrg in Ebola

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