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According to the envisioned activities of the EPIVAC project, the Final Activity Report and the document of the Disseminating the Knowledge includes the following results:
A.
Summary of the Final Activity Report.
WP1-Task1. Selection of neutralising antibodies out of a pool of HIV-specific monoclonal antibodies
Responsible Unit: Gabriela Stiegler/Polymun Scientific
Specific results :
The screening of antibodies out of a pool of 28 mAbs, directed to either gp41 (1A1, 1F11, 1H5,
2B12, 3A11, 3B7, 3D6, 3D9, 3F6, 3H12, 4B3, 4D4, 4G2, 5F3, 5G7, 6F8/5A6, 24G3, 25C2) or gp120 (1B1, 1F7, 1G2, 1G7, 2D12, 2G6, 3D12,4B1, 5G7, 3D5) of HIV-1, which were established in parallel to 2F5, 2G12 and 4E10 but not characterized.
The screening of antibodies out of a pool of 28 mAbs, directed to either gp41 (1A1, 1F11, 1H5,
2B12, 3A11, 3B7, 3D6, 3D9, 3F6, 3H12, 4B3, 4D4, 4G2, 5F3, 5G7, 6F8/5A6, 24G3, 25C2) or gp120 (1B1, 1F7, 1G2, 1G7, 2D12, 2G6, 3D12,4B1, 5G7, 3D5) of HIV-1, which were established in parallel to 2F5, 2G12 and 4E10 but not characterized.
Additionally, monoclonal antibodies towards p24 (37G12 and IAM M01 16/4/11) were produced from hybridoma cells and purified by means of a protein A capture column.
WP1-Task2. Isolation and characterisation of novel HIV-1 clade C viral isolates from recently infected subjects (to be used for the neutralisation assay).
Responsible Unit: Gabriela Stiegler
The aim of WP1-Task2 was to isolate Clade C isolates from recently HIV-1 infected donors. Originally, the laboratory of Glenda Gray (South Africa) was selected as source for samples.
Results
Four isolates (ZA349, ZA562, ZA600, ZA737) were tested for neutralisation sensitivity to C2F5, C2G12,
C4E10, the C2F5/C2G12 combination and the C2F5/C2G12/C4E10 combination.
Workpackage 2. Optimisation of vaccine delivery.
WP2-Task1. Peptide mimotopes delivered as immunogenic dendrimers of 20-60 kDa particles.
To fulfill the proposed task computational and advanced chemical methods were used, and the following activities were performed:
1. Synthesis of analogs with different substitutions and deletions, to determine the amino acids essential for binding, as well as the shortest sequence necessary for activity.
2. Synthesis and characterization of constrained analogs, through systematic variations in key positions of the sequence and introduction of molecular tools able to freeze linear and cyclic peptides in a well-defined three-dimensional structure.
Results: A set of peptides was retrieved by RPL; peptide epitopes were identified by screening the library with mAbs 2F5 and 4E10.
WP2-Task 2. Peptide mimotopes delivered as fusion products with protein carriers that stimulate both systemic and mucosal immunity.
Responsible Unit: Giuseppe Scala
Results:
A. The mimotope BS-RY (a peptide mimotope of the gp120 Bridging sheet) was folded in the context of the E2 protein, a subunit of the bacterial pyruvate deidrogenase.
In further work, sera IgG isolated from sera of rabbits immunised with the E2-BS protein-mimotope complex were shown to neutralise viral isolates, including SF162.LS, HIV.0012466-2.52 and NL-ADArs.
In a distinct set of experiments, we selected peptide mimotopes of the gp41 protein sequences corresponding to the a, immune dominant epitope;
WP2-Task3. Optimisation of the Liposome technology to deliver HIV-1 peptides and proteins
Responsible Unit: G. Stiegler
The aim of the project was the development of an efficient adjuvant based on the liposome technology.

Results:
A. DLS measurement results of two preliminary liposome preparations using PBS buffer only.
B. Liposome size (nm) for four different mimotope/liposome preparations during different preparation steps.
WP3-Task1.
Immunisation of animals with selected vaccine candidates.
Responsible unit: B. Ferko.
Results:
A. Data regarding the immunegenicity in rabbits of the liposome-peptide preparation.
WP3-Task2. Assessment of neutralizing activity of sera from immunized mice.
Responsible unit: D. Davis.
Results:
A.
Quantitative data regarding the neutralizing activity of sera from immunized animals.
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Section 2 – Dissemination of knowledge
The dissemination activities section should include past and future activities and will normally be in the form of a table maintained by the coordinator or any other person charged with controlling the dissemination activities .
Overview table
Planned
/actual
Dates
Type
Press release
Decemb er 2008
Novemb er 2008
None
Decemb er 2008
Media briefing
Conference
Exhibition
Publications
Project web-site
Posters
Flyers
Direct e-mailing
Film/video
Type of audience Countries addressed
Scientific audience Internatio nal
Higher education
Research
Industry ( sector x)
Research
None
Internatio nal
None
Internatio nal
Scientific audience Internatio nal
Size of audience
Partner responsible
/involved
Unknown Polymun
DBBM
None
200
None
None
None None
Unknown Polymun
DBBM
Unknown Polymun
DBBM
Section 3 - Publishable results and patenting

A. Identification and chemical synthesis of peptides and proteins that stimulate an anti-
HIV immune response. The novel immunogens are expected to be non-toxic.

B. When validated in monkeys models of SHIV/SIV infection, the novel immunogens are expected to share a substantial percentage of the vaccine market.

C. The project is at the development stage of validating the immune properties in rabbits; further support is required to proceed toward the non-human primate models.

Collaboration sought or offered (manufacturing agreement, financial support or investment, information exchange, training, consultancy, other)

Collaborations are welcomed from pharmaceutical companies and private/public
Institutions.

Collaborator details (type of partner sought and task to be performed)

A. The partner should be either a private –non-profit organization, or a company involved in vaccine development.

Intellectual property rights granted or published

A. The strategy and the single peptides/proteins are being covered by patenting.




Contact details: Giuseppe Scala, M.D., Ph.D.
Department of Biochemistry and
Medical Biotechnology
University Federico II
Via S. Pansini 5,
80131 Naples, ITALY
Tel: 39-081-746-3157 (lab)
Tel: 39-081-746-3622 (office)
Tel (mobile): 39-340-7760115
Fax: 39-081-746 2685
E-mail: scala@dbbm.unina.it
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