CHI's Blood Product Safety & Transmissible Spongiform Encephalopathies

Corporate Sponsors:

Human blood and plasma contain many proteins, the extraction and purification of which are of great medical and economic importance. Transmission of infectious diseases via blood transfusion and the use of processed blood plasma and components have placed a high priority on the development of new strategies for safeguarding the health of the millions of patients who receive blood-derived products every year. The screening of blood for the detection of infectious agents is continuing to advance but is complicated by new and emerging pathogens. In addition, cost-effectiveness and the threat of emerging and/or crossover infective agents must also be considered. Recent advances and new strategies for the inactivation and removal of infectious agents in whole blood, cord blood, component concentrates, and plasma are the focal point of CHI's Eighth Annual Blood Product Safety conference.

SCIENTIFIC ADVISORS
Dr. Larisa Cervenakova, American Red Cross
Dr. Jeff Davies, ZLB Bioplasma AG (Switzerland)
Dr. Fred Dombrose, Consortium For Plasma Science LLC
Dr. William N. Drohan, Clearant, Inc.
Dr. David Hammond, American Red Cross
Dr. Bernard Horowitz, Horowitz Consultants, LLC
Dr. Thomas J. Lynch, Clearant, Inc.
Dr. Paul R. McCurdy, Consultant
Dr. Stephen Petteway, Bayer Corporation
Dr. Hannelore Willkommen, Paul Ehrlich Institute (Germany)

SCREENING
Implications of NAT Testing for Transfusion Services
Economics of NAT: Does the Community Get Value for Money?
The European System for Licensing Test Kits
Screening in the 21st Century: A Regulatory Perspective
Improved Screening

VIRAL INACTIVATION
U.V. Treatment of Plasma Proteins
Gamma Irradiation of Plasma Proteins
Caprylate Mediator Inactivation of Viruses in IGIV
Inactivation of Plasma Pathogens by Pressure Cycling Technology
Effect of Broad Spectrum Pulsed Light (BSPL) on Platelet Function

NEW THERAPEUTIC PRODUCTS
Plasma Fractionation Economics: A Changing Paradigm
Reconstituted HDL: A Novel Blood Product for New Indications
Discovery of New Therapeutics from Plasma: Active Plasmin as a New Thrombolytic Agent
Manufacture and Applications of Recombinant Albumin
Plasma Proteins Derived from Transgenic Sources
Butyrylcholinesterase: Purification and Therapeutic Potentials

TISSUE SAFETY
Emerging Regulations on Tissue Safety
Ensuring the Safety of Pooled Tissue Products
Sterilization of Collagen-Based Tissue

BLOOD-RELATED SURVEILLANCE OF VCJD AND REGULATORY ASPECTS OF TSES
Update on vCJD and Blood-Related Transmission Surveillance in the UK
Surveillance Center: Present and Future
International Surveillance for Human and Animal TSEs
Regulatory Aspects of TSEs in the USA

PRION PROTEIN BIOLOGY, INTERACTIONS, AND DETECTION
The Role of Copper in the Biology of PrP
Distribution of Prion Protein (PrPc) on Blood Cells in Various Species: Speculations about PrPc Function
Sources and Properties of Prion Protein (PrPc) in Human Plasma
Interactions between Prion Protein Isoforms
Diagnosis of TSE by Cyclic Amplification of Protein Misfolding: The Prospect for a Blood Test
Screening for Prion Protein

INFECTIVITY, REMOVAL, AND THERAPEUTICS
Infectivity in Blood of Experimental Mice: vCJD Update
New Data on Blood-Borne TSE Infectivity
Inactine: Evidence of Reduction of Prions in RBC Concentrates
Drug Therapy of CJD

8:00am Registration, Poster and Exhibit Viewing, and Light Continental Breakfast

9:00 Welcome by Session Chairpersons
Dr. Jeff Davies, ZLB Bioplasma AG
Dr. Hannelore Willkommen, Head, Section of Viral Safety, Paul Ehrlich Institute

9:15 Implications of NAT Testing for Transfusion Services
Dr. Paul V. Holland, Medical Director and Chief Executive Officer, Sacramento Medical Foundation Blood Centers; and President of the International Society of Blood Transfusion
Nucleic acid technology (NAT) is being used to screen blood donations for direct evidence of hepatitis C virus and the human immune deficiency viruses that may be missed by our licensed serologic tests. Thus, blood for transfusion is safer than ever today in our hospitals. NAT will next be implemented for detecting hepatitis B viral DNA to further reduce this risk for blood being used by hospital transfusion services.
9:45 Economics of NAT: Does the Community Get Value for Money?
Dr. Michael Busch, Vice President, Research and Scientific Services, Blood Centers of the Pacific
Recent introduction of nucleic acid amplification technology (NAT) has lowered the risk of transfusion-transmitted HIV and HCV but at a significant additional cost to the health care system. We used a Markov decision model and updated estimates of viral infection risk and NAT yield to determine the marginal cost-effectiveness of NAT. The results of the base-case analysis indicated that adding pooled NAT for HIV and HCV to the current FDA-mandated battery of whole blood donation testing is expected to cost approximately $2.7 million per quality-adjusted life year (QALY) saved (sensitivity analyses yielded a range of $1.8 to $4.1 million per QALY saved). Although the cost-effectiveness of whole blood NAT is poor, political, regulatory, and public pressures to enhance blood safety appear to dictate its implementation.

10:15 The European System for Licensing Test Kits
Dr. C. Micha Nübling, Head, Section of Molecular Pathology, Division of Virology, Paul Ehrlich Institute
The European in vitro diagnostics (IVD) directive will soon replace the different current national regulations for evaluation of diagnostic kits. The European approach foresees no assessment of IVDs by a third body for the majority of products. For few products (e.g., those to be used in blood screening) the manufacturer chooses a "Notified Body" to assist in assessment of "high-risk" IVDs.

11:25 Screening in the 21st Century: A Regulatory Perspective
Dr. Indira Hewlett, Chief, Laboratory of Molecular Biology, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration
The presentation will discuss the regulatory perspectives relative to the screening of blood and blood products and update the audience with the newest information from CBER.

11:55 Improved Screening
Dr. Yanfeng Yang, Roche Molecular Diagnostics
Summary unavailable at time of printing.

2:00 Comments by Session Chairperson
Dr. Louis Aledort, The Mary Weinfeld Professor of Clinical Research In Hemophilia,
The Mount Sinai Medical Center, The Mount Sinai Hospital, Mount Sinai School of Medicine
Dr. William N. Drohan, President, Clearant, Inc.

2:10 U.V. Treatment of Plasma Proteins
Dr. Ruth Laub, Manager of Research and Development, CVBA SCRI, Belgian Red Cross
Strategy for improving biological safety will succeed thanks to advances in the pathogen screening and inactivation/removal techniques. UVC irradiation technology is a useful supplementary inactivation step in the production process to improve safety to a large range of viruses, in particular to resistant viruses such as B19. It is a reliable and inexpensive technique, completely traceable, and easy to handle and validate. Examples will be given of processes including UVC for production of fibrinogen, FVIII, albumin, etc.

2:40 Gamma Irradiation of Plasma Proteins
Dr. David M. Mann, Director of Research, Clearant, Inc.
The Clearant Process™ is a terminal sterilization process that uses high energy ionizing irradiation under radiochemically controlled conditions to inactivate pathogens in biologics of liquid or dry powder formulations. This process has been customized for a wide variety of protein therapeutics, inactivating greater than 90% of TSE infectivity or five LOGs of viruses, representing both the lipid envelope and nonenvelope types, with a minimal effect on the functional and structural integrity of the biotherapeutic.

3:10 Viral Validation, Pathogen Safety & Research, Biological Products
Dr. Kathryn Martin Remington, Bayer Corporation
A novel intravenous immunoglobulin process, based on chromatography, has been developed, which utilizes the fatty acid caprylate to precipitate lipid and lipoprotein impurities away from the product and to inactivate enveloped viruses. Under the process operating conditions, enveloped viruses were reduced to the limit of detection within minutes. In robustness studies, a wide safety margin was demonstrated with respect to relevant operating parameters. Caprylate treatment provides a useful approach for effective inactivation of enveloped viruses in biological products.

4:25 Inactivation of Plasma Pathogens by Pressure Cycling Technology
Dr. Mark Manak, Senior Vice President and General Manager, Boston Biomedica, Inc.
The repetitive application of high hydrostatic pressure at low temperatures (Pressure Cycling Technology-PCT) provides a novel approach to inactivation of blood plasma pathogens in a closed container without inactivation of the therapeutic properties of the plasma. Recent findings show that this approach is effective for inactivating both enveloped (HIV, HSV) and nonenveloped viruses (parvovirus) without damage to coagulation factors and immunoglobulins.

4:55 Effect of Broad Spectrum Pulsed Light (BSPL) on Platelet Function
Dr. William H. Cover, Vice President, Clinical and Regulatory Affairs, PurePulse Technologies, Inc.
Preliminary experimental results indicate that platelets exposed to BSPL retain activity. Exposure of platelets spiked with gram-positive bacteria to BSPL resulted in complete inactivation of the bacteria with retention of platelet function as measured by standard biochemical assays. Experimental methods developed to avoid shadowing of BSPL by the platelets are discussed.

5:45-7:00 Networking Reception (sponsored by Cambridge Healthtech Institute)

8:30 Comments by Session Chairpersons
Dr. Bernard Horowitz, Horowitz Consultants, LLC
Mr. Chris Lamb, Vice President, Plasma Operations, American Red Cross

8:40 Plasma Protein Therapeutics: A Changing Environment
Mr. Jan Bult, President, Plasma Protein Therapeutics Association
This presentation will summarize the challenges of bringing a new products to market. Challenges include financial, technical, regulatory and market acceptance.

9:10 Reconstituted HDL: A Novel Blood Product for New Indications
Dr. Peter Lerch, Head of Research, Research and Development, ZLB Bioplasma AG
This presentation will discuss biochemical, preclinical, and clinical data for reconstituted HDL.

9:40 Discovery of New Therapeutics from Plasma: Active Plasmin as a New Thrombolytic Agent
Dr. Valery Novokhatny, Senior Research Scientist II, Biological Products, Bayer Corporation
Human plasma is a unique source for production of various therapeutic proteins. Its significance as a platform for discovery of new therapeutics and their expeditious development cannot be overestimated. An example of this approach is the development of locally delivered active plasmin that is being considered by the Bayer Corporation. Data will be presented to demonstrate a therapeutic potential of plasmin for treatment of long, retracted clots found in patients suffering from limb-threatening peripheral arterial occlusion. (Coauthor: Dr. Stephen Petteway)

10:50 Manufacture and Applications of Recombinant Albumin
Dr. Werner Merkle, General Manager, Delta Biotechnology Ltd.
Summary unavailable at time of printing.

11:20 Plasma Proteins Derived from Transgenic Sources
Dr. Ian Cottingham, Assistant Director and Head of Protein Chemistry, PPL Therapeutic
Recombinant technology can provide an alternative source of plasma proteins where supply capacity or low abundance limits availability from plasma. Examples include alpha-1 antitrypsin from transgenic milk, protein C from tissue culture, and, in future, polyclonal antibodies from genetically engineered animals.

11:50 Butyrylcholinesterase: Purification and Therapeutic Potentials
Dr. Annemarie Ralston, Scientist, Plasma Derivatives, American Red Cross
Butyrylcholinesterase (BChE) is an enzyme that is naturally found at low concentrations in human plasma and whose physiological role is not known. However, BChE has been shown to rapidly inactivate cocaine and neuroblocking agents, such as succinylcholine and mivacurium. In addition, in vivo animal studies have demonstrated its efficacy as an antidote to chemical warfare nerve agents. This presentation will discuss the purification of BChE from plasma Cohn fraction IV-4 and the therapeutic potentials and limitations of BCHe for the treatment of cocaine overdose and apnea and as a preventative of organophosphate poisoning.

2:10 Emerging Regulations on Tissue Safety
Dr. Antonio Pereia, Medical Officer, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration
This presentation will outline the newest regulations governing tissue safety, with a look forward to emerging regulations.

2:40 Ensuring the Safety of Pooled Tissue Products
Dr. Randall Mills, Director of Regulatory Affairs and Technology, Regeneration Technologies, Inc.
This presentation will discuss Regeneration Technologies' program for ensuring the safety of pooled tissue products.

3:30 Sterilization of Collagen-Based Tissue
Dr. Teri Grieb, Scientist, Clearant, Inc.
Clearant has developed novel methods for the use of gamma irradiation to inactivate pathogens in therapeutic protein preparatives. These methods involve, in part, controlling the damaging effects of free radicals and reactive oxygen species generated during conventional gamma irradiation. Collagen-based tissues are particularly sensitive to the damage caused by these compounds. Current studies are designed to test the hypothesis that methods we have developed using gamma irradiation to inactivate pathogens in individual therapeutic proteins can also be optimized for the treatment of complex therapeutics such as devitalized human and animal tissues.

8:00am Registration, Poster and Exhibit Viewing, and Light Continental Breakfast

9:00 Welcome by Session Chairperson
Dr. Larisa Cervenakova, Scientist, Plasma Derivatives Department, American Red Cross

9:10 Update on vCJD and Blood-Related Transmission Surveillance in the UK
Dr. Robert Will, Consultant Neurologist, CJD Surveillance Unit, Western General Hospital
By September 2001, 106 cases of variant CJD had been identified in the UK, with an increasing incidence with time. Some vCJD cases have donated blood prior to the onset of clinical illness, but there is currently no evidence of blood-related transmission.

9:40 Surveillance Center: Present and Future
Dr. Pierluiggi Gambetti, Professor and Director, Division of Neuropathology, Case Western Reserve University
In 1997 the National Prion Disease Pathology Surveillance Center (NPDPSC) was established in the U.S. with partial funding from the Centers for Disease Control and Prevention (CDC) and the sponsorship of the American Association of Neuropathologists. The Surveillance Center' purpose is to help monitor for the occurrence of vCJD, as well as other acquired new variants, and augment surveillance for number and type of all prion diseases in the U.S. Tissues acquired are used for determining presence and characteristics of the scrapie PrP by immunohistochemistry and Western blot, which are diagnostic of prion disease, and for sequencing of the PrP gene. Cerebrospinal fluid samples are examined for the presence of 14-3-3 protein, a marker of prion disease. From January 1997 through June 15, 2001, a total of 566 U.S. cases have been examined. Three hundred and fifty cases (62%) had a prion disease. Thirty-six were familial. They included novel mutations and previously unreported phenotypes. Three cases were iatrogenic and 311 sporadic. The U.S. cases of prion disease will be reviewed along with other aspects of the activities carried out at the National Prion Disease Pathology Surveillance Center.

10:50 International Surveillance for Human and Animal TSEs
Dr. Paola Pergami, Medical Advisor, World Health Organization
At least 22 countries of the world operate surveillance systems for human TSEs resulting in important information regarding the epidemiology of humans, but particularly vCJD. Surveillance systems capable of identifying cases of vCJD require intensive resources. The WHO, in collaboration with the EuroCJD and NeuroCJD surveillance systems, is working to extend surveillance into a number of countries worldwide; these efforts and the problems involved will be described. Surveillance for BSE has undergone recent rapid transition from a passive case-reporting system to active case finding using sensitive diagnostic tests. The number of reports of BSE from countries using these systems indicates that countries that are not using active surveillance may not be finding cases even if they exist at this time. Risk assessment can guide a country as to the level of surveillance required. A review of the known epidemiology of BSE and some analysis of which countries may be most at risk will be provided.

11:20 Regulatory Aspects of TSEs in the USA
Dr. Dorothy Scott, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration
Regulatory aspects of TSEs will be discussed, including new insights into infectivity and FDA protocols to prevent the spread of these infections.

11:50 Panel Discussion with All Morning Speakers with Questions from the Audience

1:30 Comments by Session Chairperson
Dr. Byron Caughey, Senior Investigator, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health

1:40 The Role of Copper in the Biology of PrP
Dr. David Harris, Professor, Department of Cell Biology and Physiology, Washington University School of Medicine
Several lines of evidence have suggested that copper ions play a role in the biology of both PrPC and PrPSc, the normal and pathogenic forms of the prion protein. To further investigate this intriguing connection, we have analyzed how copper ions affect the biochemical properties and cellular trafficking of PrPC. Copper dramatically stimulates delivery of cell-surface PrPC to early endosomes and the Golgi and also transforms the protein into a protease-resistant form that is distinct from PrPSc. We do not confirm previous reports that the brains of PrP-null mice have a lower content of copper ions and a lower activity of Cu-Zn superoxide dismutase than normal mice.

2:10 Distribution of Prion Protein (PrPc) on Blood Cells in Various Species: Speculations about PrPc Function
Dr. Karel Holada, Visiting Fellow, Laboratory of Cellular Hematology, Division of Hematology, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration
Blood transfusion has on occasion transmitted prion diseases in experimental animals. However, large differences exist in the expression of prion protein on blood cells of experimental animals and their human counterparts. We speculate that those discrepancies may serve as a protective function against transfusion transmission of human prion diseases. (Coauthors: Jan Simak and Jaroslav G. Vostal)

2:40 Sources and Properties of Prion Protein (PrPc) in Human Plasma
Dr. Ian MacGregor, Lead Scientist, Products and Components Research Group, National Science Laboratory, Scottish National Blood Transfusion Service
Application of a DELFIA immunoassay for PrPc shows that a major portion of the total pool of human blood PrPc resides in the plasma. Studies on isolated megakaryocytes, platelets, and endothelial cells indicate that they all contain and can release PrPc. However, measurements of circulating PrPc in bloods from patients with a range of haematological disorders lead us to conclude that vascular endothelium may be the chief source of plasma PrPc. Grossly elevated levels of plasma PrPc in patients with end-stage renal failure suggest that renal catabolism may be an important route of clearance, and this may be of relevance to recent reports of an abnormally folded isoform of PrP in urine of patients suffering from prion diseases.

3:50 Interactions between Prion Protein Isoforms
Dr. Byron Caughey
Important biological aspects of TSE diseases such as strains and species barriers appear to be controlled, at least in part, by specificities in the interactions between various normal and TSE-associated forms of PrP. Studies of these interactions have also provided insight into the mechanism of the conversion between the normal and protease-resistant forms. New classes of inhibitors of the PrP conversion have been identified that may serve as lead compounds in the search for anti-TSE therapies.

4:20 Diagnosis of TSE by Cyclic Amplification of Protein Misfolding: The Prospect for a Blood Test
Dr. Claudio Soto, Senior Scientist and Head, Department of Molecular Neuropathology, Serono Pharmaceutical Research Institute
"Protein Misfolding Cyclic Amplification (PMCA)" mimics in the test tube the process of prion replication in a "fast-forward" mode. Undetectable amounts of PrPSc are incubated with a large excess of PrPC to allow the conversion of the normal protein. The process is amplified by cycles of sonication in order to multiply exponentially the amount of PrPSc, simplifying its detection. PMCA has enormous potential in allowing current diagnostic tools to detect BSE and vCJD at a much earlier stage, even during the presymptomatic period, and may open the door to detecting the disease in living people using blood or peripheral tissue samples. In addition, PMCA represents a good tool to help understand the underlying biology of prions, to identify other factors implicated in prion protein conversion, and to discover novel drug targets for prion diseases.

4:50 Industry Strategies for Addressing CJD Risks
Mr. Chris Healey, Executive Director - North America, Plasma Protein Therapeutics Association
The presentation will identify steps taken by the plasma therapeutics industry to address stakeholder concerns and understanding of the risks CJD may present to plasma therapeutics.

5:20 Panel Discussion with All Afternoon Speakers with Questions from the Audience

5:45-7:00 Networking Reception (sponsored by Cambridge Healthtech Institute)

9:00 Comments by Session Chairperson
Dr. David Hammond, American Red Cross

9:10 Infectivity in Blood of Experimental Mice: vCJD Update
Dr. Larisa Cervenakova
Previously, we reported on an experimental study in mice to investigate the presence and distribution of vCJD infectivity in their blood. The study is in progress. To date, the disease has been transmitted upon intracerebral and intravenous inoculation of mice with buffy coat collected from vCJD-infected animals during the incubation period and at the clinical phase of the disease. Intracerebral inoculation of plasma from clinically ill animals has transmitted the disease as well. The levels of infectivity in blood components tested for vCJD have not been greater than the levels of infectivity identified for the Fukuoka-1 strain.

9:40 New Data on Blood-Borne TSE Infectivity
Dr. Robert Rohwer, Director of Molecular and Neurovirology, Veterans Affairs Medical Center-Baltimore
We will present the results of four large studies: (1) the titer of blood-borne BSE infectivity in the 301v mouse model, (2) the dynamics of the TSE infection of blood during the preclinical phase of infection, (3) the effect of inoculation route on blood-borne TSE infectivity, and (4) the contribution of spleen and spleen cells to the TSE infectivity of circulating white blood cells.

10:50 A Novel Approach to improve Product Safety: the Gradiflow as a tool to remove Prions and pathogen
Dr. Hari Nair, CEO,Gradipore Inc.
Currently available separations technologies for pathogen removal are limited and there has been a global emphasis on a search for a new technology which can both enhance product yield and safety. An important aspect of product safety centers on the ability of separation technologies to remove prions.Recent studies have shown that a novel membrane-based separationtechnology, the Gradiflow, has the potential to provide a solution bysuccessfully purifying proteins while concurrently removing prions andboth enveloped and non-enveloped viruses from blood/plasma. Thissession will provide an overview of this new separation technology and its applications for prion, endotoxin and pathogen removal.

11:20 Drug Therapy of CJD
Dr. Paul W. Brown, Laboratory of Central Nervous Studies, National Institute of Neurological Disorders and Stroke, National Institutes of Health
Attempts to treat transmissible spongiform encephalopathies began over 30 years ago and have yet to register a significant success. However, recent studies have emphasized the importance of a chemical structure containing both water-soluble and lipid-soluble components capable of interacting with the membrane-bound proteinase-resistant PrP (ÔprionÕ protein). A number of compounds shown to eliminate PrP and/or infectivity in scrapie-infected tissue cultures are the subject of ongoing studies in animals and are under consideration for human drug trials. As with other recalcitrant infections, combinations of drugs with different modes of action are likely to be necessary for any effective therapy. Also, very recent work in developing antibodies that can neutralize in vitro infection (and, in conjunction with genetic engineering, in vivo infection) has renewed interest in the strategy of prophylactic vaccination.

11:50 Panel Discussion with All Morning Speakers with Questions from the Audience