Meetings 1998

Minutes of the Meeting of the Representatives research laboratories and institutions working with UNESCO in the fields of AIDS research and prevention
Regional Office for Science and Technology for Europe (UVO-ROSTE)
UNESCO Venice Office 5-7 September 1998, Venice, Italy

The Representatives research laboratories and institutions working with UNESCO in the fields of AIDS research and prevention met on 5-7 September 1998 at the UNESCO Office in Venice, Italy.

Members who attended the meeting were:

BECKER, Prof. Yechiel
UNESCO - Hebrew University of Jerusalem, International School for Molecular Biology and Microbiology, Dept. of
Molecular Virology, Faculty of Medicine 91120 Jerusalem, Israel tel. +972-2-675-8394, fax. +972-2-678-4010 Email: becker@md2huji.ac.il
BERETTA, Dr. Alberto
Research Director CIRBS, 185 ave. Robert Scauman 75014 Paris, France tel. +33-1-4412-3183, fax. +33-1-4412-3270 Email: aberetta@worldnet.fr
BORISSOV, Mr. Boris
Deputy Secretary General Russian Commission for UNESCO 9 Vozdvizhenka Ave.Moscow 121019 Russian Federation tel. +7-095-291-001, fax. +7-095-202-1083
COLIZZI, Prof. Vittorio
University of Rome "Tor Vergata" Dept. of Biology Via della Ricerca Scientifica 00133 Rome, Italy tel. +39-6-7259-4237, fax. +39-6-7259-4224 Email: colizzi@univroma2.it
COSTA, Dr. Cristina
Documentation Center on AIDS "IRCCS" - "L. Spallanzani" International Center on AIDS and Other Emerging Infections Via Portunese 292 Rome 00149 Italy tel./fax. +39-6-5517-0430, Email: unesco@uniroma2.it
FORTI, Prof. Augusto
Senior Special Advisor to Director General of UNESCO for Europe and North America UNESCO 7 Place de Fontenoy 75352 Paris France tel. +33-1-4568-1431, fax. +33-1-4568-5552
GONZALEZ - CANALI, Dr. Gustavo
CIRBS Hopital St. Joseph 185 rue Raymond Losserand 75014 Paris, France tel. +33-1-4412-3181, fax. +33-1-4412-3270 Email: ustavog@cirbs.worldnet.fr
GOUGEON, Dr. Marie Lise
Chief of Laboratory Institut Pasteur 28 rue du Dr. Roux 75724 Paris Cedex 15 France +33-14568-8907, fax. +33-1-
4568-8909 Email: mlgouges@pasteur.fr
IPPOLITO, Dr. Giuseppe
Scientific Director National Institute for Infectious Diseases Via Portuense 292, 00149 Rome,Italy tel. +39-6- 559
4223,fax. +39-6-559-4224 Email: craids@excalhq.it
KADIO, Prof. Auguste
Faculty of Medicine B.P. V 166 Abidjan, Cote d'Ivoire tel. +225-255-249, fax. +225-442-897 Email: kadioa@syfed.refer.ci
KOUZMINOV, Dr. Vladimir
Deputy Director UNESCO Venice Office 1262/A Dorsoduro 30123 Venice, Italy tel. +39-41-522-5535, fax. +39-41-528-9995 Email: roste@unesco.org
MONTAGNIER, Prof. Luc
President World Foundation for AIDS Research and Prevention c/o UNESCO 1 rue Miollis 75352
Paris France tel. +33-1-4568-4599, fax. +33-1-4568-8916
MORONI, Prof. Mauro
University of Milan Clinic Infectious Diseases L. Sacco Hospital Via G.B. Grassi 74 20100 Milan, Italy tel. +39-02-356-0010, fax. +39-02-356-0809
POCCIA, Dr. Fabrizio
National Institute for Infectious Diseases International Center on AIDS and Other Emerging Diseases and Re-emerging
Infections Via Portuense, 292-00149 Rome, Italy tel. +39-6-55170430, fax. +39-6-5517-043Email: poccia@uniroma2.it
SPIRIN, Prof. Alexander
Institute of Protein Research, Russian Academy of Sciences Pushchino 142292 Russian Federation tel. +7-095-924-0493, fax. +7-095-924-0493 Email: protres@sovam.com
THIBODEAU, Prof. Lise
Institut Armand Frappier (INRS) University of Quebec 531 Boulevard des Prairies Laval, P.Q. CanadaH7N 4Z3 tel.+1-450-687-5010, fax. +1-450-686-5640 Email: lisethibodeau@iafuquebec.ca
VAGLIANI, Mr. Pierluigi
Executive Secretary General World Foundation for AIDS Research and Prevention 1 rue Miollis 75015 Paris, France
tel. +33-1-4568-4520, fax. +33-1-4273-3745
ZEMBALA, Prof. Marek
Jagiellonian University Medical College Dept. Clinical Immunology Wielicka 265 Cracow 30663 Poland tel. +48-12
658-2486, fax. 48-12-658-1756 Email: mizembal@kinga.cyt.kr.edu.pl

Report of the Panel of Representatives of the Research Laboratories and Institutions working with UNESCO in the fields of AIDS research and prevention.

   The Panel or Representatives positively evaluated the projects concerning the study of HIV Nef protein directed by Prof. L. Montagnier involving several groups of the network as follow: Prof. A.S. Spirin, Prof. L. Thibodeau, Prof. V.Colizzi and Dr. F.Poccia. Prof. Montagnier presented several observations on a possible role of Nef protein in polyclonal T cell activation. Specifically, recombinant nef proteins were shown to be able to significantly increase T cell activation induced by anti-CD3 antibodies. These evidences support a striking role of nef protein both in activation of the immune system and in promotion of viral infection spread. This and other evidences in the literature support the idea of a nef-based vaccine approach. Prof. Montagnier and Prof. Thibodeau are studing the effects of neutralizing antibodies against nef and possible applications of liposomes. In particular, Prof. Thibodeau is producing large amount of nef protein in baculovirus system. In parallel, Prof. Spirin proposed a cell free system for the production of large amount of nef protein. In this laboratory it has been demonstrated that continuos protein synthesis over long periods resulting in high protein yield can be attained in cell-free systems, when the incubation is performed not under static conditions in a fixed volume but in the flow of a feeding solution through the cell-free extract. The continuous-flow cell-free (CFCF) systems mentioned can be generally used for preparative syntheses of the various viral antigens that are toxic for living cells. The synthesis of Nef, is proposed to accomplish in the CFCF bioreactor, with the object of its use for AIDS diagnostics and antibody induction. The production of large amount of nef protein in baculovirus and CFCF system is recommended by the committee, who promotes a exchange of these reagents between the groups working in this field. Prof. Montagnier will promote and direct the sharing of reagents and informations. Prof. Montagnier also reminded that aver since its creation, "Man against viruses" is committed to the development of new vaccines. He underlined the necessity of exploring several approaches such as gp160, Nef, Tat, conserved sequences in C5, and/or Nef in association with gp160 or C5. He also stressed the opportunity to explore vaccinotherapy for immune reconstitution in seropositive patients, under treetherapy, or not. Dr. Poccia and Prof. Colizzi proposed a project concerning the analysis of the T helper response to HIV-nef protein in order to develop new vaccination strategies. The main objective will be to evaluate the extend of a protective anti-nef T helper response in HIV infected persons who control viremia in the absence of antiviral therapy or in patients treated with potent antiviral therapy. Specifically, the project concern: a) analysis of T helper anti-nef responses and epitope mapping; b) analysis of nef- polymorphisms and/or deletions of nef sequences; c) evaluation of a nef- pathogenic index in viral isolates. This study will provide preliminary informations on the feasibility of a nef-based vaccinotherapy in HIV-patients undergoing potent antiviral therapy. The development of this project has been positively considered by the Panel of Representatives. Dr. Thibodeau mentioned that she is currently constructing a Nef-deleted HIV-LAI virus that could be useful as a control and offered to provide us with such a virus.

   The committee approved the continuation of the research project of Dr.A.Beretta concerning the study of exposed uninfected individuals in Paris (CIRBS) and Abidjan (CIRBA). This project shall provide the key for the development of tests of HIV suppressive T cell immunity. The test that will be developed within this first phase of the study are: a) a test of "relative resistance to Infection" of PBMC (at least two different types of cloned viruses will be used: CCR5 and CXCR4 dependent viruses; the profile of relative resistance to in vitro infection of ExU individuals PBMC will be also evaluated); b) a test of in vitro production of HIV suppressive activity (CAFlike activity) (we have generated a large number of CD8 cell clones from two ExU individuals; we should soon be able to identify the HIV suppressive activity produced by these clones; we are also devoting a particular effort to understand the immunoregulation of the production of these factors); c) a test of peptide C5 specific immunity: proliferative index, production of chemokines.

   Prof. L.Thibodeau presented briefly her research program on the development of an HIV-Immunosome subunit vaccine against HIV and underlined the importance of an immune response at the genital surface to achieve protection from the sexual transmission of the virus. She presented recent results where it was shown, for the first time, that immunization of rabbits by the oral/parenteral/mucosal routes (using a minimal protocol of three interventions) induces antibodies in sera, tears, saliva, vaginal and intestinal secretions, capable of neutralizing primary isolates. The second part of her talk concerned the strategy she has developed to overcome the immunodominance of gp120 V3 loop. To achieve this goal, a recombinant gp160 was engineered, in which the immunodominant V3 loop was deleted, except for the GPGRAF tip of the loop, which is highly conserved (gp160- GPGRAF). A second construction, in which the V3 loop was completely deleted ( gp160 V3+) was also made. The modified protein, gp160- GPGRAF, retained its CD4 binding properties, whereas, the gp160 V3+ failed to bind the receptor. The gp160- GPGRAF was expressed in a baculovirus system, purified by chromatography, formulated as an Immunosome (gp160-coated liposome) and used to vaccinate mice. Neutralization assays against six divergent laboratory strains and against six primary isolates clearly showed that a deletion of part of the V3 loop, keeping the GPGRAF tip, favors the induction of broadly reactive neutralizing antibodies. Such a construction and the ability of the modified protein to induce high titers of neutralizing activity, not yet reported in the literature, brings new hope towards the development of a protective vaccine against HIV infections. This vaccine will now be used for oral/ parenteral/mucosal immunizations in order to determine its potential of inducing broad spectrum neutralizing antibodies and T cells response at the genital and rectal levels. The members of the assembly evaluated positively the results and underlined the large amount of work that was done. The Committee approved the continuation of the mucosal immunity approach using the HIV-1 V3-modified gp160, to be tested in rabbits, until funds are available to evaluate this vaccine in macaques.

   Prof.Y. Becker presented his project: "Synthetic Peptides and DNA Vaccines against HIV-1 for Application to Skin Langerhans Cells (HIV-1 Peplotion Vaccine) to induce Antiviral Cytotoxic T cells". This project will concern: a) studies on the conditions for intraepidermal transfer of synthetic peptides to skin Langerhans cells; b) computer analyses of HIV-1 proteins to identify cleavable nonapeptides for use in HIV-1 Peplotion Vaccine; c) the development of HSV-1 Peplotion Vaccine to immunize mice a model system; d) a proposal to initiate studies on the vaccination of AIDS patients (volunteers) with HIV-1 Peplotion Vaccine. Two approaches have been proposed. The first step can be done in collaboration with AIDS clinic members of Man Against Virus Network. Blood dendridic cells may be obtained from HIV-1 infected volunteers. Specifically, the Panel promoted the collaboration with the International Center for AIDS and other Emerging and Re-emerging Infection of Rome on the development of in vitro assay to characterize blood dendritic cells functional activity with Prof.V.Colizzi and Dr.F.Poccia. The autologous dendritic cells will be incubated in vitro, under suitable conditions, with synthetic HIV-1 peptides, compatible with the HLA classI haplotypes of each patient. The synthetic peptide treated autologous dendritic cells will be reinjected in to each volunteer and at different time intervals, anti HIV-1 CTLs in the peripheral blood of the volunteer will be monitored. The virus load and anti-HIV-1 antibodies content in the blood sample from the volunteer will provide a measure for the usefulness of the synthetic peptide vaccines. Finally, HIV-1 infected volunteers will be immunized with an HIV-1 Peplotion Vaccine applied to the area of the skin that was treated by therapeutic ultrasound. At different time intervals blood samples will be taken from each volunteer to monitor virus content, anti HIV-1 antibodies and CTLs activity. The Panel support of Prof. Becker 's project and raccomends preliminar in vitro studies on dendritic cells.

   The Panel approved the activities performed by the UNESCO Interdisciplinary Chair in Biotechnology held at the University of Rome "Tor Vergata" by Prof. Marek Zembala. The creation of this Chair is the new initiative of UNESCO in agreement with the University of Rome "Tor Vergata". The Chair was established in a close cooperation with the local biomedical research and training units. The Rector of the University decided that during academic year 1997/1998 the Chair was associated with the Department of Biology, Laboratory of Molecular Pathology, headed by professor Vittorio Colizzi. The Teaching activities was held to run the course on "Immunotechnology in Biology and medicine". The course was run in English with introduction and summary given in Italian. The course was designed to give the updated information on the recent advances in immuno-and bio-technologies in research and medicine with special attention on future directions. The course was not medically orientated but examples of some human diseases were used to illustrate the progress and application of immunotechnology. It covered the application of modern immunotechnologies in three broad fields: cancer, inflammation and infection as applied for diagnosis, prophylaxis and treatment of human diseases as well in biomedical and pharmaceutical research. At the end of the course detailed practical information was provided on how to use certain techniques, e.g. cell phenotyping, sorting, purging, isolation and detection by various immunological methods. A positive opinion of the panel was also expressed on the research activities directed by Prof. M.Zembala concerning a study on Human monocyte / macrophage response to HIV-1 and tumours. Monocyte / macrophage are involved in AIDS pathophysiology as they are major targets for HIV-1 in vivo, from an important reservoir of the virus and are responsible for the virus spread to tissues. The study performed indicates that coinfection in vitro with Mycobacterium tuberculosis leads to an increased HIV-1 replication in monocytes and it enhanced transmission to T cells. Several phenotypic and functional changes of monocytes have been observed in HIV-1+ subjects: depressed antigen presenting capacity, both decreased and increased HLA -DR and enhanced CD11 a expression. Some of these changes that are pertinent for immunopathogenesis of AIDS may be induced by HIV-1 envelope gp120. The use of monocytes / macrophages either infected with HIV-1 or stimulated with gp120 is planned to study their response to cancer cells in vitro and to determine their effect in vivo on tumor growth in SCID mice.
The Panel approved the support of the reserch innovative project presented by Dr. F.Poccia: "Immunotherapy with nonpepptidic antigens in SIV-infected macaques". Previous studies have shown that gd T lymphocytes a) are the most potent killers of HIV/SIV infected cells and b) become anergic in many asymptomatic HIV-infected persons. gd T cells are important 'first-line defense players' contributing substantially to the initial control of the viral infection, but their chronic activation may help (probably through the cytokine release) to maintain the persistent overactivation of the immune system, which in turn results in immune dysfunction associated with AIDS. This project proposes to test whether vaccines containing potent nonpeptide antigens for gd T cells will influence both the control of SIV replication and the maintainence of immune functions in in vivo infections of rhesus monkeys (Macaca mulatta). The efficacy of the in vivo and the in vitro testing will be analyzed and documented. Successful outcomes may lead to selecting novel vaccination and immune reconstitution strategies for clinical testing in human AIDS.

   The Panel positively considers the institution of the "International Center for AIDS and other Emerging & Re-emerging Infections" directed by Prof. V. Colizzi, which originated from the Agreement between the National Institute for Infectious Diseases "Lazzaro Spallanzani", UNESCO and the World Foundation for Research and Prevention of AIDS. Institutional sponsorships have been expressed by the Italian Ministry of Health, by the National Research Council and by the University of Rome "Tor Vergata". The general objective of the International Center will be to develop the scientific research and its divulgation in the fields of AIDS and other emerging and re-emerging infections, with special attention paid to promoting the integration of the International Center with the Laboratories and Clinical Divisions of the National Institute for Infectious Diseases "L.Spallanzani", and joining with networks activated in this field by the UNESCO and by other UN Agencies. The International Center was recently inaugurated on March the 25th 1998 and is organized in the following three functional units :
1. Research Laboratory. The Laboratory is the operative structure in the research field carrying out the research projects identified by the CNR-UNESCO Committee, the 'L.Spallanzani' Institute and the World Foundation. The Laboratory building has been functionally set-up and equipped by 'L.Spallanzani' Institute with instruments supplied by UNESCO, the University of Rome 'Tor Vergata'. In this first stage of activation, the staff of Laboratory is made up of personnel of 'L.Spallanzani' Institute, university scientists, and by UNESCO and "Istituto Superiore di Sanità" scholarships.
2. Technical Services. The scientific activity of the Research Laboratory will integrate the Laboratories and Clinical Divisions of the Spallanzani Institute by setting up technical services for training and basic research development, including: flow cytometry and image analysis, nucleotide sequencing and bank of biological samples.
3. Documentation and Scientific Communication Service. Aim of the Service is to collect and catalogue the divulgation materials produced by UNESCO and other national and international agencies, and to promote the scientific communications of the International Center by organizing meetings, scientific expositions, Internet web pages and so on. The Service will work in close collaboration with Volunteer Associations working within 'L.Spallanzani' Institute.

   The Panel approved the proposal of Dr. G.Ippolito concerning the future activities of the UNESCO Center at the Spallanzani Institute in Rome, as follow:

- The UNESCO Center will be integrated in the new department of experimental research. The new building will be opened in October 1999 and will have a P3 laboratory.

- The UNESCO Center will receive part of new technical equipment, obtained with a special grant from the Ministry of Health to the National Institute for Infectious Disease " L. Spallanzani".

-The integration of UNESCO Center in the current research activities will allow to this center to have a steady support by "L. Spallanzani" Institute.

-The "Spallanzani" Institute is hardly working to increase his scientific power and visibility in basic research and to improve clinical and experimental activities in which has a natural leadership. The UNESCO Center represents a central part of this groove.

-The Institute Spallanzani is promoting activities in the field of HCV treatment, behavioral aspects of infectious disease and of information and documentation on HIV prevention and response, including advertising, campaign and specific literature. The role of UNESCO in the customs of a documentation center is central.

The Spallanzani is working at international level in the field of emerging and re-emerging infection with special attention to the infectious disease in the Mediterranean area. The Italian public health authorities are concerned for the spread of communicable diseases in the Mediterranean Countries. This spread is in part related to the wars in the Eastern Countries and to displaced peoples. The Spallanzani conducts in the last years interventions for surveillance and control of communicable diseases in several countries under construct of WHO and other international agencies. This experience and relevant results obtained allow to identify L. Spallanzani Institute as an ideal place for the creation of an European Center for communicable diseases in the Mediterranean area in favor of activities promoted or supported by European Union.


   A positive evaluation was expressed by the Panel on the scientific promotion and collaborations presented by the Laboratory directed by Prof. M.L.Gougeon. Specifically, it as been noted the relevance of the BIOMED programs performed under the financial support of the European Union in the past years. The results obtained by these collaboration, involving several French and Italian groups are witnessed by several scientific reports. These studies ranges from T cell anergy to apoptosis. Specific contributions were done in the field of the mechanisms of apoptosis in AIDS an on the functional activity of gd T lymphocytes. A scientific program of the European Union called Copernicus is actually directed by Prof. M.L.Gougeon. This program concern the role of apoptosis in the depletion of non infected CD4 T lymphocytes in HIV infection. The objectives of the project are to develop new techniques for the detection of apoptosis, to correlate these techniques with the onset of HIV infection and test the potential therapeutical effect of retinoids on the apoptosis of HIV-infected lymphocytes.

   The panel postively evaluate the proposal of Prof. A.Kadio concerning the institution of an international chair in the field of Education, Science and Culture located at Abidjan. This Chair has the main objective to promote the transfer of knowledge and to help the developing countries to consolidate their formation and research ability. The course on formation on HIV/AIDS was created in collaboration with the University of Abidjan. This course concerns the medical, social and economical aspects concerning AIDS in Africa. In the last century, while pharmacological therapies and vaccination practices greatly reduced the microbial infections diffusion in developed countries, in most Third World countries infectious diseases remain the main cause of death. For example, AIDS related pandemy favored the development and the spread of drug-resistant M.Tuberculosis strains; in some African countries, tuberculosis is the first death cause among HIV-infected patients. In developed countries, on the other hand, C hepatitis incidence is very high among HIV patients, and is associated with a marked increase of hepatic damages and an inefficacy of available therapies. The hospitals are unable to take care of the increasing number of patients. Thus, the employment of additional people/services is necessary. However, specialized personnel is required to take care of AIDS patients. This underlines the need of personnel formed in handling the of medical and social AIDS-related problems. The actual political, economical and cultural situation favored the expansion of AIDS. Thus, the creation and organization of a formation center on HIV and related retroviruses at Abidjan is recommended.

   Dr. V.Kouzminov headlined the role of the UNESCO UVO Office in the coordination of the activities of the representatives research laboratories and institutions working with UNESCO in the fields of AIDS research and prevention.
Dr. Kouzminov underlined that UNESCO will favour exchange of scientific information, reagents, new and innovative research techniques between members of the network "Man against viruses", in order to share the common "know how" and reinforce the links between the members. Moreover, it has been confirmed the purpose of UNESCO in providing support to new initiative in this fields and stressed the relevance to extend the financial support of the newborn initiative/project to the national and international appropriated financial agencies. Nevertheless, the UNESCO will continue to play its natural role in the starting up of initiative and or projects concerning the activities of research laboratories and institutions working with UNESCO in the fields of AIDS research and prevention.

The Panel recommends:

· the promotion and development of the research projects on HIV-1 LAI Nef protein to be used as a vaccine and as a reagent to study some aspects of HIV-induced pathogenesis, directed by Prof. L.Montagnier, who will promote and direct the sharing of reagents and informations between the research laboratory involved (Prof. L.Thibodeau, Prof. A.S. Spirin, Prof.V.Colizzi and Dr.F.Poccia). The continuation of the cloning, production and purification of HIV-0 VAU Nef protein by Prof. L. Thibodeau's laboratory for similar studies.

· the continuation of the research project of Dr. A.Beretta concerning the study of exposed uninfected individuals in Paris (CIRBS) and Abidjan (CIRBA)

· the continuation of the mucosal immunity research project of Prof. L.Thibodeau on HIV-subunit vaccine, in particular the comparison of recombinant GPGRAF-Immunosome with undeleted gp160-Immunosome in regard to their potential for inducing systemic and mucosal immune response capable of neutralizing divergent laboratory strains an also monocytropic and T-lymphotropic primary isolates.

· the development of the research project of Prof. Y.Becker: " "Synthetic Peptides and DNA Vaccines against HIV-1 for Application to Skin Langerhans Cells (HIV-1 Peplotion Vaccine) to induce Antiviral Cytotoxic T cells". Moreover, the Panel promoted the collaboration with the International Center for AIDS and other Emerging and Re-emerging Infection of Rome on the development of in vitro assay to characterize blood dendritic cells functional activity with Prof.V.Colizzi and Dr.F.Poccia.

· the continuation of the UNESCO interdisciplinary Chair in Biotechnology at the University of Rome "Tor Vergata".

· take in consideration the results obtained through the network by the different Centers located in the Mediterranean area. The Panel is concerned for the diffusion of communicable diseases in the Mediterranean area. This spread is in part related to the wars in the Eastern European Cauntries and to movement of population of displaced peoples. The Panel evaluates the need of an intervention targeted to evaluate scientific, clinical, social and human right aspects.

· the creation at the Spallanzani Institute of an European Center for Research, training and information on Committee in the Mediterranean Area in the framework of activities of the European Union, considering that there is a National Institute for Infectious Diseases in Rome with clinical, scientific and social experiences. Moreover, the Spallanzani Institute in Rome conducted in the last years interventions in the Mediterranean area in connection with WHO, International Center for Infection and Health, International Center for Migration and Health, WHO Europe and European Union.

· the continuation of the collaborative projects directed by Prof. M.L.Gougeon concerning T cell anergy and apoptosis in HIV infection.

· the proposal of Prof. A.Kadio concerning the institution of an international chair in the field of Education, Science and Culture located at Abidjan.

· that UNESCO, in close cooperation with World Foundation for AIDS Research and Prevention, will continue to play its natural role in the starting up of initiative and or projects concerning the activities of research laboratories and institutions working with UNESCO in the fields of AIDS research and prevention.