Planning Meeting for the NIAID Malaria Research
and Reference Reagent Repository
November 24-25, 1997
The planning meeting for the NIAID Malaria Research and Reference Repository began with a welcome by the two chairpersons, Drs. Mary Galinski and John Adams. The welcome was followed by a brief reminder of the meeting's purpose, which was to gather ideas from investigators involved in various aspects of malaria research concerning how the new malaria repository should function. NIAID will consider the recommendations of this group in further development of plans for the repository.
Dr. Stephanie James, Chief of the Parasitology and International Programs Branch, NIAID, thanked the participants for coming to the meeting and restated its purpose - to focus on input from investigators directly involved in malaria research to determine what needs should be addressed by the new repository. Dr. James opened the general discussion by reviewing the history of the Multilateral Initiative on Malaria, and presenting an overview of current NIAID activities in malaria research including the establishment of a malaria research and reference reagent repository.
The Multilateral Initiative on Malaria is a loose federation of international research, disease control, and development assistance agencies with interests in malaria control. The goal of the MIM, as stated at a meeting held in April, 1996, is "to strengthen and sustain, through collaborative research and training, the capability of malaria endemic countries in Africa to carry out research required to develop and improve tools for malaria control." At a subsequent meeting held in Dakar in January, 1997, the issue of creating reagent repositories was raised. Specifically, scientists representing multiple aspects of malaria research (pathogenesis, epidemiology, entomology, immunology, antimalarial drugs, health systems and operational research, interventions, mechanisms of cooperation and support) stated a need for:
1) improved access to parasite, vector and human reagents; and,
2) standardization of assays using well-characterized and renewable reagents.
NIAID has attempted to respond to these stated needs of the malaria research community by establishment of a malaria reagent repository for the use of qualified scientists worldwide. The Institute has created an interim facility for one year (FY 1998), during which time the mechanism will be put in place to make a seven year contract award for a full repository.
Description of the Interim Repository
Dr. Michael Gottlieb, Program Officer, Parasitology and International Programs Branch, NIAID, described the interim repository and the responsibilities of the interim contractor, who was present at this meeting. The contractor is:
Dr. S. Ved Brat
Braton Biotech, Inc.
1 Taft Court, Suite 101
Rockville, MD 20850
The interim contract was issued in late September, 1997 and will continue until September, 1998. This contract will not perform all of the functions expected of the full contract. Interim contractor responsibilities include:
1) acquisition of reagents from collaborating scientists;
2) provision of facilities for hybridoma expansion and rabbit immunizations;
3) development of quality assurance procedures;
4) coordination of storage, inventory and distribution to approved investigators;
5) development, maintenance and utilization of an inventory and tracking database system; and
6) purchase and utilization of necessary equipment for storage. The interim repository contract provides support for acquiring (including shipping) and generating reagents.
The recipient of repository reagents will be responsible for shipping costs. The following reagents will be included in the interim repository:
1) hyperimmune sera;
2) monoclonal antibodies;
3) recombinant proteins and antigens; and
4) molecular biology reagents, including libraries, clones, constructs and probes. No parasites, mosquito vectors or human sera/tissues will be included in the interim repository.
Dr. Gottlieb enumerated the expected benefits for contributors to the repository:
1) easing the burden of distribution for those investigators who already supply reagents to others;
2) potential to generate new reagents; and
3) increased access to renewable reagents by the malaria research community.
Description of Full Repository
Dr. James described the RFP (Request for Proposals) for a "Malaria Research and Reference Reagent Repository" (RFP-NIH-NIAID-DMID-98-17), as well as an RFP for "Malaria: Clinical Research and Trial Preparation Sites in Endemic Areas" (RFP-NIH-NIAID-DMID-98-19) which contains provision for collection of field-acquired materials for access through the repository. The proposal receipt date for both RFP's is January 23, 1998. The RFP for the reagent repository contract will be subject to full and open competition and proposals will undergo the standard NIH review process. Award of the repository contract is expected before the end of FY 1998.
Dr. James outlined the additional aspects of the full repository, which will expand on the capacity of the interim repository. The full repository will include access to Plasmodium parasites, vector reagents, human sera and human tissues. As a part of this contract, a scientific advisory committee (SAC) will be established to provide ongoing advice on issues such as prioritization of reagent acquisition, regular review of progress, etc. Capacity to provide reagent quality analysis and control (QA/QC) will also be provided, to relieve donors of the bulk of this burden. In addition, the repository will serve to provide public information and will support workshops and training activities.
Charge to the Planning Group
Dr. James gave the charge to the participants of this planning meeting. The purpose of this planning meeting was to find out more specifically what is needed by malaria researchers so that the repository can supply materials accordingly. She stated that the intent of the repository is to provide malaria reagents to the entire research community, not just NIAID-supported investigators.
She emphasized that participants were asked to serve as representatives of their various areas of interest in malaria, not as individual scientists, to advise NIAID on needs and concerns of the community regarding:
1) reagent needs and prioritization;
2) characterization requirements and quality control;
3) intellectual property rights and ethical issues;
4) training and technology transfer; and
5) publicity for the repository.
Status of Ethics Discussions on Use of Human Materials within Repositories
Dr. Roger Aamodt, Chief of the Resources and Development Branch, NCI, NIH, presented an overview of ethical considerations regarding the use of human materials in research. Dr. Aamodt serves on the Trans-NIH Working Group on Human Subjects and Tissue Resources. He began with a list of issues related to the use of human tissue. These include:
1) autonomy and beneficence;
2) risk to patient;
3) risks to investigators involved in the use of human tissue;
4) ownership of tissues and medical data;
5) purposes of use;
6) commercialization; and
7) what the tissue donor receives as compensation.
Dr. Aamodt pointed out that use of human tissue without prior consent has been typical in the past. At the present time, advance directives or blanket consent may be used for a patient population.
A number of questions have arisen concerning ethics of use of human tissue:
1) Does each and every use of stored tissue require specific consent each time it is used?
2) Should subjects be recontacted without their consent?
3) How can subjects be informed of results obtained using their tissue?
4) Should a subject be informed when results of experiments using their tissues are directly applicable to their health?
5) Exactly what is the definition of research involving living human subjects?
The federal government has established the Office for Protection from Research Risks (OPRR) to provide guidance on protection for human subjects in research. Institutional Review Boards (IRB's) have been established to review federally funded human subject research to be certain that the investigator is complying with federal regulations involving protection of human subjects. Localities/institutions may also have established IRB's, but where the research is funded by the US government these IRB's must also conform to US federal regulations, regardless of the state or foreign country of origin. Informed consent or waiver of informed consent is the approach required by IRB's.
OPRR subjects repositories of human tissue to oversight by an IRB. Use of tissues stored at these repositories involves no direct contact between tissue donors and investigators who wish to use the tissue for research purposes. The repository acts as a "broker" between the two in the following manner:
Tissue Collector - -> Repository Storage & Data Management Center - -> Recipient Investigator
Such an arrangement assures the anonymity of the tissue donor. Both the tissue collector and the repository operator must be reviewed by an IRB and comply with IRB recommendations. Informed consent must be obtained for each sample sent to the repository, unless the IRB finds that the research involves only minimal risk to the donor.
Genetic studies are becoming increasingly popular, and with them come complicated issues regarding use of human genetic material. Genetic privacy laws have been passed to protect individuals from insurance, health and employment discrimination based on genetic predisposition to certain conditions. There are currently 16 bills pending legislation in the US Congress, and another 100+ bills in at least 34 states in the last year have dealt with this important issue. In general, unless conditions for limitation or waiver are met, informed consent must be obtained for genetic studies involving humans. It has been agreed that the benefits to society must outweigh risks to the donor. The critical nature of patient privacy is pointed out by the formation of the National Bioethics Advisory Commission by President Clinton in 1996. A Genetics Subcommittee report expected in January/February, 1998, will publish guidelines for the use of stored existing and prospective human genetic materials. Any potential contractor should be prepared to keep abreast of developments in this area.
Function of the DAIDS Repository
Dr. Opendra Sharma of the Division of AIDS Research (DAIDS), NIAID presented an outline of the AIDS Research and Reference Reagent Program, which has served as a model for the proposed repository of malaria research reagents. The DAIDS repository contract was first awarded in 1988, beginning with 60 reagents, and was recompeted in 1993. A catalog is now published annually, containing over 2500 reagents, including 135 AIDS virus isolates, biologically active clones, recombinant peptides and proteins, drug resistant virus samples and cell culture viral samples. Also included in the DAIDS catalog are reference panels of monoclonal antibodies, polyclonal antibodies, PCR probes for HIV envelope genetic subtyping and drug resistant viruses. In addition, human body fluids containing virus/antibodies and organisms of opportunistic infection (in AIDS patients) are available. The current breakdown of reagents is approximately 22% virus samples, 16% antibodies and 27% recombinant clones. Repository users (registrants) include 60% from academia, 9% industrial and 10% government research. At this time, 39 countries are using reagents from the DAIDS repository.
The DAIDS reagent program has accomplished much since its origin in 1988. It represents a unique resource to the scientific community through which small amounts of reagents are supplied free of charge to requestors. The repository also dispenses information and technology transfer through publication of a newsletter, which contains updates of protocols and announcements of research initiatives to its recipients. New reagent classifications have resulted from commercial developments by the repository, and workshops have been conducted, e.g., Heteroduplex Mobility Assay and Shipping of Infectious Materials. The repository has been cited in over 1300 scientific publications. Two reagent kits have been produced for distribution: a Heteroduplex Mobility Assay kit and an HIV-1 p24 detection kit.
Dr. Sharma briefly reviewed the Registration Form used by the DAIDS repository (included in the catalog). Each requestor must complete forms (and obtain institutional officer[s'] signature[s]) regarding such information as compliance with safety standards, shipping company account number for billing purposes, agreement for use in animals only except where permission is obtained prior to request, reporting agreement, assumption of shipping costs, and agreement for indemnification of the US government, the contractor and reagent donors. In addition, the requestor must send a biographical sketch or CV.
The Registration Form also contains an agreement concerning commercial discoveries resulting from the use of a reagent from the repository. Five Release Categories have been established for reagent classification for promotion of commercial development:
A) Recipient may use reagent without prior agreement ~35% of recipients
B) Recipient shares income with donor institution ~20-25% of recipients
C Recipient signs agreement for commercial use prior to receiving reagent ~20-25% of recipients
D) Recipient selects category B) or C) prior to receiving reagent~20-25% of recipients
E) Recipient must not use or incorporate reagent for commercial purposes.
Sometimes a donor delegates only one time use per requestor, after which the requestor must contact the donor for permission to use the reagent again; certain institutions (e.g. biotech companies) consider all requests as having commercial potential, assuming there is no such thing as research without commercial prospects.
Recipients are responsible for payment of shipping charges. Dr. Sharma noted that 70% of reagents are requested by NIH-funded investigators. He also pointed out that shipping reagents to overseas sites is often a problem because of difficulties in obtaining approvals for shipping from the countries to which reagents are to be sent. This may present a problem for the malaria repository, since many research sites are located outside the US.
Dr. Haldar raised the subject of how complaints to the repository were dealt with. Dr. Sharma indicated that these cases were rare, but pointed out one specific example of a reagent recall in which all recipients were sent a letter of apology by the contractor. He also stated that it sometimes turns out that the complaining requestor hasn't followed the protocol recommended by the donor or contractor as carefully as he/she should have.
Dr. Krishna asked whether most donations to the DAIDS repository are solicited or unsolicited. Dr. Sharma replied that donations are largely unsolicited, due to the great extent of advertisement of the repository at major meetings and by the reagent reporting system. New products are constantly being announced.
Dr. Taylor asked if there are any particular requirements that must be met in order to donate reagents to the repository. The answer was that once a new reagent has appeared in a publication, donors are encouraged to send samples to the repository. When a reagent is sent by a donor to the repository, a reagent form (stating how it was produced and a protocol for its use or a brief description of how the reagent may be used) accompanies the shipment.
Dr. Leke asked what can be done to ease shipping problems to African countries, where time required for shipping and high costs can create prohibitive concerns for both donors and recipients of repository reagents. Dr. Sharma explained that the January, 1995 "tightening" of infectious agent shipping regulations by the US Department of Transportation and Air Transportation has loosened somewhat in succeeding years. The use of liquid nitrogen shipments has improved the safe transport of infectious materials from areas as far away as Siberia, Estonia and Malawi, in the case of AIDS reagents. Shipping problems which still remain to be solved include shipping for arrival on weekends or holidays and the policy that no infectious materials are allowed in the airspace over India. Dr. Sharma pointed out that the NIH Office of Safety works closely with DAIDS to ensure shipping safety.
Other queries had to do with storage of the repository materials once they are received. In the case of the DAIDS repository, all storage facilities are located in the U.S., but satellite laboratories could potentially be opened in other countries for temporary storage, especially when a country wishes to send only part of its supply of a reagent to the central repository. Discussants agreed that such satellites would be particularly useful in the case of malaria, since most disease occurs in Africa and shipping materials to/from Africa is often complicated. Satellite repositories could also be used for reagents belonging to investigators who are unable to continue to support a project, but do not want to dispose of reagents collected under that project because of their potential utility to others.
Presently, the DAIDS repository collaborates with the MRC, UNAIDS, WHO and the UK. The MRC reagents program is smaller than the DAIDS repository, but MRC has unique virus reagents which could be made available to the research community. MRC publishes a small catalog of reagents which are shared with the DAIDS Repository. UNAIDS is responsible for isolation of field isolates of the AIDS virus. WHO assists developing nations, especially where officials are unaware of safety regulations for infectious materials.
As an addendum to these discussions, Dr. Collins of CDC pointed out that a special permit is needed for blood samples from endangered species (e.g. certain species of monkeys), which will be something to remember when the full repository is up and running.
Formation of Interest Groups for Discussion
The following breakout groups were formed, and the following spokespersons chosen: Vaccines and Immunology - Dr. James Kazura; Vector Biology - Dr. Frank Collins; and Molecular and Cell Biology - Dr. John Dame. Interest groups were asked to discuss major issues related to the formation of the full repository (from their group's perspective), according to eight criteria:
Criteria for prioritization of reagent acquisition/development
Minimum requirements for reagent characterization upon donation to the repository
Minimum requirements for QA/QC for different types of reagents
Distribution mechanism for field-acquired reagents
How to handle proprietary concerns
How to deal with ethical issues regarding use of human tissues
Publicity/solicitation of reagents
Group Discussion Following Breakout Groups
Following the breakout session, each interest group presented responses to the above list of issues for further discussion by the reconvened group.
Combined Recommendations from Molecular and Cell Biology/Vaccines and Immunology Focus Groups: (combined because of overlap in several areas)
I. Prioritization of reagent acquisition/development.
A. General Recommendations
B. Make available "state of the art" technologies when they arise
- Include reagents for perceived needs of the malaria community
- Follow recommendations of the Advisory Committee
- Conduct surveys and maintain interactive database
- Work to contact remote areas
C. Assess regularly whether the needs of those with limited resources are met
- Include research and training in endemic areas
- Include satellite repositories for distribution in endemic countries
- Priority should be given to include items with public health impact in endemic countries
- Country specific issues should be tied in with MIM activities
II. Priority parasite species
A. Human malaria
B. Mouse malaria
- Plasmodium falciparum
- Plasmodium vivax
- Plasmodium ovale
- Plasmodium malariae
C. Monkey malaria
- Plasmodium berghei
- Plasmodium yoelii
- Plasmodium chabaudi
D. Avian malaria
- Plasmodium knowlesi
- Plasmodium cynomolgi
- Plasmodium fragile
III. Priority for obtaining reagents (not ranked in order)
- Plasmodium gallinaceum
A. Antigens & Antibodies
B. Parasite lines and clones
- Recombinant proteins as Reference antigens
- Monospecific antisera
- Monoclonal antibodies
- Diagnostic reagents/ Immunogen-specific identification assay
- Pooled serum from endemic countries, pooled by age group, parasite species
- Individual serum with phenotype information attached, but encrypted
C. Antimalarial compounds
- Lab maintained
- From humans from various areas with different histories
- From animal tissues
D. Molecular typing kits
- Drug sensitivity assays
- Drugs with restricted availability
- Kits - for training component especially in endemic areas
IV. Minimum requirements for reagent characterization upon donation to the repository.
- Parasite DNA
- Gene libraries - genomic, cDNA, YAC
- Characterized molecular gene clones
- Transformation vectors
- Oligonucleotide primers
- łKits" - for training component especially in endemic areas
- Chip" technology when available
A. DNA clones and vectors
B. Gene libraries
- Restriction map
- Sequence tag
C. Parasite lines
- Assay for host contamination (or other contaminants)
- Insert size
- Primary or amplified
D. Field acquired parasite isolates
- Geographic origin
- Clinical profile
- Isolation history
- Passage history
- Cloning method
- Genotype & Fingerprint (e.g., MSP-1, MSP-2, Glurp - 3 & other polymorphic loci)
- Test for mycoplasma
- Geographic location (GPS coordinates if possible)
- Date of recovery
- Description of study
- Phenotype - if known from study
- Clinical status of the patient - treatment history and clinical outcome
- DNA - fingerprinting for genomic
- Individual serum with phenotype information
- Patient phenotype attached, but encrypted
F. Monoclonal antibodies
- Immunogen or antigen source for immunization and titer assay
- Animal and strain
G. Recombinant immunogens, peptides
- Immunogen or antigen source for immunization and titer assay
- Selection process
- Antibody isotype
V. Minimum requirements for QA/QC for different types of reagents
- Sequence & Strain
- A "checklist" for immunological characterization
- Immunogen-specific identification assay
A. HIV and Hepatitis B testing of parasite isolates and other samples containing human blood or sera
B. Test for Mycoplasma contamination and other identified problem microbial contaminants
C. See RFP for others
VI. Distribution mechanism for field-acquired reagents
A. Some form of lab certification for receipt of materials
B. Restrict shipment of transformed or drug resistant parasites to an endemic area; they should not be sent to endemic countries under any circumstances
VII. How to handle proprietary concerns
A. Adopt DAIDS model
B. Donor assigns release requirements
C. Acknowledgment of individual donor as well as the malaria repository when publishing data based on materials received from the repository
VIII. How to deal with ethical issues regarding use of human tissues
A. Ethical issues were covered adequately by Dr. Roger Aamodt of NCI
B. Contractor will need to keep apprised of developments in this area.
IX. Training/technology transfer
A. Satellite repositories for training purposes - capacity building and serve as local source in endemic area
B. Training offered by repository personnel on use of reagents and kits
X. Publicity/solicitation of reagents
A. Word of mouth among workers in the field
C. Scientific meetings
D. Repository staff should participate in those meetings
E. Staff should offer workshops/training in "hot" new methods and use of reagents
F. Web sites
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