(Onderstepoort, South Africa)

24^th-25^th April 2002

1. First 12-monthly scientific report required in Brussels by 31^st January 2003.

Please send directly to Philip Mellor at IAH-Pirbright to arrive no later than 31^st December 2002. I will then consolidate your contributions into one comprehensive scientific report & will submit it to Brussels.

2. First 12-monthly cost statement required in Brussels by 31^st January 2003. Please send directly to Philip Mellor at IAH-Pirbright to arrive no later than 31^st December 2002. I will pass these on to Mr Andrew Grice (Institute Secretary) or Mr John Mottram (Institute Finance Officer) who will consolidate your statements & submit them to Brussels. Should your own finance officer anticipate difficulties in completing your cost statement then s/he should contact Mr Mottram directly (tel. 00 44 1635 578 411, fax 00 44 1635 577 237, e-mail john.mottram@bbsrc.ac.uk).

3. After acceptance of the scientific reports & cost statements the Commission will allocate the second tranche of money (approx 22.5% of the total) within 2 months to IAH.

4. The anticipated rates of financial allocation from the Commission are as follows:

40% advance.

22.5% at end of 1^st year after acceptance of the relevant scientific reports & cost statements.

22.5% at end of 2^nd year after acceptance of the relevant scientific reports & cost statements.

15% after acceptance of the final reports & cost statements.

 

1. WP 1 - To attempt to establish laboratory colonies of potential European vector species of Culicoides (e.g. C. imicola, C. obsoletus gp, C. pulicaris gp, C. nubeculosus) for use as a tool in BT epidemiological studies.

PARTNERS involved in this area of work:

Partner 2 (OVI) - Responsible partner

Partner 1 (IAH)

Deliverables/tasks wholly or partially scheduled for the 1^st year of the project:

By month 24 - Methods for the collection/rearing of 1 or more potential vector Culicoides species devised. Blood feeding in the laboratory achieved & eggs deposited in simulated breeding sites by Partners 1 & 2.

By Month 36 - Methods for the rearing of eggs through to adults of 1 or more potential vector Culicoides species devised & provisional colonies established by Partners 1 & 2.

What has been achieved so far in these areas & with which species?

Following Mr Meiswinkel's departure who will lead this work at Partner 2?

Which species should we concentrate on in the remainder of the first year of the project?

What do we know about the breeding sites of these species?

What should/do we need to do during the remainder of 2002 to ensure achieving our set deliverables?

PARTNERS involved in this area of work:

Partner 1 (IAH) - Responsible partner

Partner 2 (OVI)

Deliverables/tasks wholly or partially scheduled for the 1^st year of the project:

By Month 12 - Sufficient quantities of all required monovalent vaccine viruses (serotypes 4, 9 & 16) produced by partner 2 & supplied to partner 1.

Two of these viruses to have been passaged at least once (intrathoracically & orally) through vector insects, inoculated into sheep & clinical signs recorded by Partners 1 & 2.

By Month 24 - All 3 vaccine viruses to have been passaged at least once (intrathoracically & orally) through vector insects, inoculated into sheep & clinical signs recorded by Partners 1 & 2.

By Month 36 - All 3 vaccine viruses to have been passaged twice (intrathoracically & orally) through vector insects, inoculated into sheep & clinical signs recorded by Partners 1 & 2.

What has been achieved so far in these areas of work?

Do we have sufficient amounts of the necessary monovalent vaccine viruses to do the work?

What potential vector species of Culicoides should Partner 1 concentrate on (obsoletus, pulicaris, nubeculosus)?

Do we have a reliable scoring system for the clinical signs of BT elicited in sheep, so that the results of partners 1 & 2 can be made directly comparable?

What should/do we need to do during the remainder of 2002 to ensure achieving our objectives?

PARTNERS involved in this area of work:

Partner 1 (IAH) - Responsible partner

Partner 2 (OVI)

Deliverables/tasks wholly or partially scheduled for the 1^st year of the project:

By Month 12 - Selection of suitable parental strains of BTV for reassortment studies by Partners 1 & 2.

By Month 24 - Initial characterisation of the parental (wild type & vaccine) strains of BTV by Partners 1 & 2.

By Month 24 - Carry out dual infection experiments in insect & mammalian hosts, & in cell culture, followed by plaque purification of progeny viruses by Partners 1 & 2.

What if anything has been done so far in this area of work?

Have the suitable parental strains yet been selected & if they have what are they?

What stage are we at, in terms of the initial characterisation of the parental strains?

What is the projected timetable over the first 2 years of the project for carrying out the dual infection experiments?

Are any interactions with other partners, visits, etc required to facilitate this work?

4. WP 4 - To develop efficacious inactivated whole virus vaccines, to measure their immunogenicity in comparison with the existing live virus vaccines & existing experimental sub-unit vaccines.

PARTNERS involved in this area of work:

Partner 3 (MERIAL) - Responsible partner Partner 1 (IAH)

Partner 2 (OVI) Partner 4 (GVS)

Partner 5 (LSHTM) Partner 6 (LSPHA)

Deliverables/tasks wholly or partially scheduled for the 1^st year of the project:

By Month 1 - Field isolates of BTV types 2, 4, 9 & 16 supplied to Partner 3 by Partners 1 & 4.

By Month 9 - Master Virus Seeds of BTV types 2, 4, 9 & 16 produced by Partner 3.

By Month 12 - Growth & purification characteristics of selected European BTV types determined by Partner 1.

By Month 15 - Large volumes of BTV suspensions produced by Partner 3.

By Month 16 - Method validated for quantification of the antigen by Partner 3.

By Month 17 - Methods validated for inactivation of the 4 types of BTV by Partner 3.

By Month 23 - Production of industrial batches of concentrated purified BT antigen by Partner 3.

By Month 24 - VLPs (& CLPs?) of EU BTV types/strains generated for evaluation as a basis for inactivated vaccines by Partner 5.

By Month 24 - Slow release formulations generated by Partner 6 using expressed particles, inactivated native particles & individually expressed BTV proteins provided by Partner 1.

What if anything has been done so far?

Do we have sufficient examples of each of the European BTV field strains (2, 4, 9 & 16) for Partner 3's needs, if not can Partners 1 & 4 supply what is required?

What is the timetable for transferring the viruses to Partner 3? How should they be transferred? What other information does Partner 3 require about these viruses?

What is the projected timetable of Partner 3 for:

Selecting suitable candidate vaccine strains from the above collection & for preparing Master Seed Vaccine strains?

Determining & measuring the parameters for BTV culture on BHK cells, complete inactivation, & concentration & purification of BTV suspensions?

Initiating the work set out in WP 4.3?

What is the projected workplan & timetable of Partner 5 for generation of the required amounts of VLPs (& CLPs?) & what is required from other Partners to carry out this work?

What is the projected workplan & timetable of Partner 6 for the generation of the slow release formulations & what is required from other Partners to carry out this work?

Any other areas for discussion?

5. WP 5 - To develop & evaluate synthetic particle or subunit-based vaccines using recombinant baculovirus expressed VLPs, CLPs &/or other orbivirus proteins, & to develop & test assays differentiating between infected & vaccinated animals.

Partner 5 (LSHTM) - Joint responsible partner Partner 1 (IAH) - Joint responsible partner

Partner 2 (OVI) Partner 4 (GVS)

Partner 6 (LSPHA)

By Month 18 - Generation of cDNA clones of outer capsid protein genes from BTV types/strains by Partners 1, 2 & 5.

By Month 24 - Generation of VLPs of BTV types/strains, as well as CLP & individual BTV proteins for evaluation as vaccines by Partners 1 & 5.

By Month 30 - Evaluation of recombinant expressed BTV proteins/particles as vaccines by Partners 1, 2 & 4.

By Month 36 - Generation of slow release formulations using expressed particles, inactivated native particles or individually expressed BTV proteins by Partners 1, 2, 5 & 6.

By Month 36 - Development of an assay to distinguish between vaccinated & BTV infected animals by Partners 1, 4 & 5.

Suggested reas for discussion:

Generation of cDNA clones etc.

Partner 1 is contracted to isolate & grow the BTV strains/types involved in the European outbreaks (2, 4, 9, 16) & maybe 1, 6, 10 & 13 to provide live virus or genomic dsRNA to Partners 2 & 5 in order to produce baculovirus expressed VLPs - what has been done so far? What needs to be done to fulfil our contractual obligations within the specified time scale (18 months)? In the light of the departure of Albie van Dijk & Janusz Paweska, can Partner 2 still participate in this area of work?

Generation of VLPs of BTV types/strains, as well as CLP & individual BTV proteins for evaluation as vaccine.

Partners 5 & 1 are contracted to produce sufficient amounts of these reagents "to permit a comparison of their efficacy in comparison with the existing vaccines & with other inactivated vaccines". So what amounts of these reagents will be required in theory & what amounts is it possible to produce in practice?

Generation of slow release formulations using expressed particles, inactivated native particles or individually expressed BTV proteins by Partners 1, 2, 3, 5 & 6 (Project Task 13). What is required by Partner 6 from Partners 1, 2, 3 & 5 to initiate and complete this work & in what time scale?

If carried out comprehensively this would be a colossal & expensive area of work involving between 5 & 8 BTV serotypes, (each as VLP & individual outer capsid protein formulations with & without slow release delivery systems, all in comparison with the inactivated whole virus vaccines & the existing live vaccines).

So realistically what is the minimum/best that we can do? Should we concentrate on only a "selection" of the virus serotypes involved, if so which ones? What is the minimum number of sheep that we can use in the comparative testing of each formulation in order to provide meaningful results? - Advice please from Partners 2, 3, 4, & 5.

Live virus vaccine + whole inactivated particle + VLP + individual protein(s) + (each with & without slow release) = 8 to 10 + controls + selected dosage rates.

How many & what dosage rates should we use with VLPs, individual proteins etc?

Perhaps Partners 2 & 4 should develop a joint testing protocol for the work to be carried out in sheep so that results are comparable?

Any other thoughts on how we can most effectively handle this area of work?

Development of an assay to distinguish between vaccinated & BTV infected animals by Partners, 1, 4 & 5.

What has been done so far in this area of work?

What else does each of the involved Partners need to do to achieve the development & evaluation of this assay (i.e. what approach should we adopt, what reagents are required & how can they be acquired)?

What is the anticipated time scale?

Any other points?

6. WP 6 - To develop molecular epidemiological methods to identify the source of BTV's causing outbreaks (in Europe) & to differentiate between vaccine & field strains of BTV.

PARTNERS involved in this area of work:

Partner 1 (IAH) - Responsible partner

Partner 2 (OVI)

Partner 4 (GVS)

Partner 5 (LSHTM)

By Month 24 - Generation of RNA sequence data for BTV strains that represent an immediate threat to Europe (1, 2, 4, 6, 9, 10, 13, 16) by Partners 1, 2, 4 & 5.

By Month 36 - Establishment of an RNA sequence database to help identify the origins of BTV incursions into Europe & to enable differentiation between vaccine & field strains of virus by Partners 1, 2, 4 & 5.

What has been done so far in this area of work?

Have Partners 2 & 4 provided Partner 1 with all of the required BTV isolates available to them? What are these isolates (i.e. species of origin, geographical location of isolation, date of isolation, passage history, etc)?

What is the projected time scale of Partners 1, 2 & 5 for the sequence analysis & comparisons between these viruses?

Could Partner 1 provide an update on the current situation regarding the establishment of a BTV RNA sequence database to enable strain identification & differentiation? How may other Partners access this information?

Any other points?

To attempt to colonise European potential vector species of Culicoides in the laboratory & to assess reversion to virulence of vaccine viruses in sheep subsequent to passage through one or more of these vectors (possibly C. imicola, C. nubeculosus, C. obsoletus).

To examine the ability of live vaccine viruses to re-assort with wild-type viruses in both insect vectors & vertebrate hosts & to partially characterise any reassortants.

To develop methodologies for producing safe, efficacious, inactivated whole virus vaccines & to measure their immunogenicity in comparison with the existing live virus vaccines.

To carry out trials comparing the efficacy of experimental sub-unit vaccines with new inactivated whole-virus vaccines & the commercially available live virus vaccines, & to evaluate them as candidates for commercial production.

To further develop & evaluate particle or subunit-based vaccines using recombinant baculovirus expressed virus-like particles & core-like particles of BTV or based upon other Orbivirus proteins (e.g. VP7 or VP2).

To develop test(s) enabling differentiation between animals infected with a live BTV (field or vaccine virus) infection & those vaccinated with non-replicating vaccines.

To develop molecular epidemiological methodologies for backtracking BTV outbreaks to source & differentiating vaccine viruses from field strains.