Global Networking Against Malaria
NIAID Malaria Research and Reference Reagent Repository
Recommendations from the Vector Biology Interest Group:
Perceived needs and general comments:
40-50 mosquito colonies are currently available, including ~30 strains of An. gambiae. Aedes species should not be included.
The group agreed that establishing the colonies within the repository should not be delayed
Backup colonies should be available at a different site
Quality control will be the responsibility of the subcontractor(s)
Number of insect colonies, especially Anopheles, required will rise with time as transgenic lines are established
Specimens of non-colonizable (field collected) specimens should be stored as well
Kits and probes
Species identification of cryptic species
Infection status of field specimens collected
Clones, libraries, etc. (See list above from Vaccines and Immunology/Molecular and Cell Biology groups)
Arrayed libraries of Anopheles gambiae on microtiter plates (BAC) are available - the group questioned whether these could be provided.
g) Other services/training needs:
Infection of mosquitoes
Use of kits and probes
h) Research on cryopreservation
Responses to the Eight Issues
By historical use
ELISA kits for species identification, clones
2/3. QA and QC
Generally the same as DAIDS
Mosquito colony QC by subcontractor
QC should be done only by someone with experience in this area.
Currently, there are only a few insectories with sufficient experience.
Distribution of live mosquitoes
Send to no part of the world (i.e., endemic countries) where they could potentially extablish and create a transmission problem
PHS approval for receipt
Safety/ethical issues must be considered
Oversight by insectory manager (subcontractor) and contractor
Collaborating institutions could serve as sources
Field caught mosquito specimens
Both Project Officer and specimen donor should have input into distribution decisions
Proprietary concerns - none
Training/technology transfer (see above list)
Publicity - AnoDB (Anopheles Data Base)
Discussion followed Dr. Kazura's presentation for the Vaccines and Immunology Focus Group. Dr. Galinski lead with a question concerning storage of archival material - is it appropriate to keep archival human sera from past studies in case someone should want to reexamine it; how is it classified? Dr. Ved Brat, interim repository contractor, explained that human serum can be classified either as a human product or or reagent, depending upon whether or not identifiers are available. Identifiers for human material tracing it back to a particular individual would necessitate its classification as a human product. A sample with no link to an individual would be classifiable as a reagent. A set of samples linked to a particular ethnic group, parentage (general) or sex would also be classified as reagents (but this narrows the classification somewhat).
Solutions for assurance of confidentiality of donor tissues were also discussed, including:
1) encryption of individual data;
2) informed consent at the start of a study, HIV testing (since the presence of HIV can confuse serologic testing data) - Dr. Leke pointed out that in areas of low HIV transmission in parts of Africa people don't really believe that HIV exists and may refuse testing for that reason, whereas in areas of higher HIV transmission people tend to cooperate more readily in being tested;
3) banks of human sera could be pooled from vaccine trials, rather than storing individually labeled serum samples. Serum from vaccine studies, either failed or successful, can be difficult to store because of stability problems.
Dr. Sharma indicated that sharing with contributors information on what is being done with reagents that are donated to the repository keeps a spirit of openness and assures contributors that their contributions are valuable.
The remainder of the meeting was largely given over to general discussion about the recommendations of the three interest groups. This discussion coalesced around three major topics.
Discussion began regarding whether the repository would be supplying reagents or reference materials, since reagent production is very costly. Dr. Dame wondered what happens when a supply of a reagent is exhausted - does the repository go back to the donor or produce more material at the repository? Drs. Brat and Sharma commented that the purpose of a repository is not to produce large quantities of anything, but to produce ėgold standardî reference material for comparison of the quality of another reagent. Large volume scale-up is too costly and time consuming a venture for the repository to assume. The group agreed that, up to a point, the repository would have to make certain that enough of a reference reagent would be available to fill requests, but that the intent of the full repository is not to produce large amounts of a given reagent (e.g., amounts sufficient for longterm experiments or clinical trials). Instead, the repository would send a ėseedî amount of a particular reagent to a requestor, and it would be that requestor's responsibility to produce the amount needed to complete the work or form a collaboration with the original donor if greater quantities are required.
Dr. Dame asked whether there were a separate budget recommended for support of each area (e.g., parasite strains, clones, monoclonals, etc.); Carl Henn, of NIAID Contracts Management Branch (CMB), replied that the scientific advisory committee will be established when the full repository contract is awarded to help to select funding priorities. This planning meeting was convened to start the process, since the committee will not be in effect during the year of the interim repository.
Suggestions for priority reagents, including parasites and insects, were then made. Dr. Walliker reminded everyone that WHO is a source for some parasites; he felt that DNA and PCR kits would be a priority for the NIAID repository. Dr. Collins of CDC indicated that he presently supplies researchers with a large number of mosquitoes, mosquito eggs, human blood, and human serum by shipping from his CDC laboratory to requestors, and he sees this as a probable ongoing demand from the repository. Dr. James asked the group whether previous suppliers of specific malaria reagents should be entirely relieved of these responsibilities by the new repository, or if previous suppliers should remain in charge of their respective reagents while the repository functions to fill in the current gaps in reagent supply. Almost unanimously, the group agreed that they would be glad to turn over these types of duties to the repository, which would allow them more time for their own research. Producing bulk reagents, vialing them, ėQCîing, etc., is very time consuming. Dr. Sharma agreed, stating that the DAIDS repository has taken over supplying reagents which were formerly sent out by individual labs, and the repository sends back a report to the previous supplier regarding the processing of their reagent(s). It was agreed that the repository would not take over functions which others are willing or able to continue in terms of supplying reagents to the research community.
Dr. Sharma reminded everyone that the key to a repository is providing ėStandard Reference Reagentsî. When asked if the DAIDS repository had ėleveled outî in popularity, Dr. Sharma responded that about 10,000 samples/year are requested and supplied, proof that the AIDS research community relies on the repository as intended. The group agreed that the establishment of ėgold standardî reference reagents, immunogens and monoclonal antibodies is absolutely critical. This means that a single source should be considered for each type of reagent, and similar reagents subsequently acquired through expansion or donations should be compared to that ėgold standardî, so that investigators will be performing assays using the same standard of comparison.
Dr. Leke asked the group what would be the repository's expectation of people in endemic regions. Dr. Adams addressed the question by stating that probably investigators in endemic countries would be expected to donate parasite isolates, human tissues (blood, serum, etc.) from population studies, and other reagents which may be new markers for pathogenesis, drug resistance, etc. In turn, these investigators could expect, through the repository, to gain accessibility to typing reagents, reference sera, monoclonal antibodies, genomic DNA and primers for diagnostic PCR, as well as training in the use of various assays. In other words, the benefits would be reciprocal. Dr. Leke added that P. ovale and P. malariae are occurring more frequently in Africa and that standardized monoclonal antibodies against these two species should be added to the repository along with those against P. falciparum and vivax.
Regarding maintenance of insect colonies, Dr. Collins reiterated that the vector group felt insectories are a priority which should not be delayed until the third year of the contract and wondered whether this could be accommodated. Mr. Henn of CMB replied that a pre-award amendment to the contract can be made to establish an insectory component before the third year. NIAID program staff stated that they would look into the possibility of obtaining additional funding for earlier startup of vector production by the full repository.
For advertising the repository throughout the malaria research community, the group consensus was that the primary methods should be:
1) to announce its formation at major tropical disease meetings, such as the upcoming American Society of Tropical Medicine and Hygiene meeting in December; and,
2) to spread the word through various scientific newsletters. It will be more difficult to reach those investigators involved in field research in endemic areas. E-mail is said to reach about one half its intended recipients, so the other half would have to be contacted by regular mail. Information sent to all attendees at the January Dakar meeting would potentially reach those in the field.
Dr. Galinski suggested that the Malaria Foundation's web site and specifically its Malaria Research Network and WorldWide Malaria Directory could be utilized to inform investigators about the repository and encourage them to contribute reagents and participate in the development of a functional repository. She offered her assistance in engaging the global scientific community in this process through utilization of these electronic resources, which can be found at: http://www.malaria.org.
Dr. Oduola suggested that the three breakout groups reconvene to take their initial lists and discuss additions of reagents which are of particular interest to the respective groups, keeping in mind the subjects of ėdonationî and ėreceiptî as priorities for discussion. Breakout groups were again formed and were asked to generate two lists: one of prioritized (potential) donations and the other of prioritized reagents that are likely to be requested as soon as they are available. At the end of the breakout period the groups reconvened to discuss deletions/additions to the previous list of issues. These will form the basis of a ėsurveyî to be circulated to the malaria research community to begin the process of reagent acquisition and repository utilization. Everyone agreed that a one page introduction explaining the purpose of the repository should be added to any list which is circulated throughout the malaria community. There was agreement that a list of participants of this meeting should also be circulated, so that investigators with an interest in the repository have contact information should they have questions or suggestions.
Dr. Kazura reported an update from further discussions of the Vaccine Interest Group. It was decided that thinking should be redirected toward the development of a repository which is particularly accessible to investigators and clinicians from endemic countries, since it is generally more difficult for them to obtain research materials. Of particular importance would be ėGold Standardî monoclonal antibodies, recombinant antigens and primers (or genotyping). These would be especially useful if they were available in assay kits which could be used by endemic scientists to identify parasite species or measure immune responses of infected individuals.
A discussion followed concerning the inclusion of reagents generated as a result of unpublished data. Varying individual opinions were expressed regarding releasing such reagents for general use, with the result that the group agreed that the ėcomfort zoneî of donating laboratories should be respected and that individual labs should decide whether or not to include unpublished materials. There was general agreement, however, that reagents which are used as tools to accomplish a specific task could easily be made available to the repository even though they are unpublished and may never be (e.g., constructs made as part of an experiment which was not published).
Dr. Leke suggested that, in addition to making reagents available, it would be useful for the repository to sponsor workshops and training in the use of these reagents in the endemic countries. She added that training in DNA extraction and cryopreservation would be especially helpful. Dr. Gottlieb reminded the group that discussing training activities will be part of the scientific advisory committee's duties when the full repository is established. In addition, Dr. James stated that some of the endemic country training is included in the RFP for Clinical Research and Trial Preparation Sites in Endemic Areas (RFP-NIH-NIAID-DMID-98-19) and will be covered by that contract. In addition, some of the recently awarded Fogarty International Center training supplements contain malaria training initiatives. Thus, the repository will not be totally responsible for this aspect.
Dr. Gottlieb summarized the priorities of donor information to be supplied to the repository along with each contribution:
1) the nature of the reagent;
2) the method(s) of expansion or production of the reagent;
3) type and results of quality control measures;
4) storage and distribution requirements and restrictions;
5) aliquot type (vial, dried pellet, etc.) and amounts. recommended; and
6) references in publications, if any have resulted at the time of donation.
The meeting was adjourned with the reminder that this represents a beginning which will be expanded upon as the repository plans develop, and increasing numbers of investigators add both their ideas and reagents.
[Note added after the meeting: As recommended, the RFP has been modified to include insectory facilities in the first year of the contract. NIAID is using lists generated by the planning group to prepare a survey of malaria research reagents which will be placed on the Institute home page in January.]