Research

Researches

Activity Report

Annual Report 2008

Download the Annual Report 2008

Executive Summary
The Research Programs implemented at HKU-PRC are dealing with infectious diseases that may have devastating effect on public health and the economy of the community. HKU-PRC has a particular niche to fill in bringing together the cutting edge developments in the cell-biology of infectious disease, which is the strength of Institut Pasteur, with the expertise in the clinical virology and immunology of infectious disease, which is the strength of HKU. The general strategy of the new directorate team, which had been approved by the Board of management in February 2008, has been presented to the Scientific Advisory Board, which met for the first time in October 2008. The overall evaluation was very positive and the SAB was impressed with the excellent start in establishing a high-quality research enterprise to take advantage of the integrated research programs of HKU and Institut Pasteur, as well as with our strong international and complementary network of collaborators. During 2008 we have been able to achieve significant milestones in the development of the center. In addition, HKU-PRC has been strengthening its teaching program and is actively involved in leading local and regional network projects.

We have solidified our research focus on respiratory infections and on the mosquito borne disease of dengue by securing several independent extramural grants, thus validating the scientific orientation and the experimental approach that are being pursued. These programs are strengthened through several key collaborations with Institut Pasteur, HKU, and the Pasteur International Network. The scientific output of the Center is high, with more than 20 manuscripts published since 2007 (10 in 2008) and several publications having already appeared in 2009 or being in the final stages before submission. The Center is an integral part of the Area of Excellence (AoE) on Influenza and has a number of research programs related to this program. The Scientific Director, Professor Peiris serves as the coordinator of this multi-departmental and multi-institutional program.

The fundamental direction of HKU-PRC emphasizes interdisciplinary approaches to confront the challenges posed by viruses and we wish to embed this orientation by linking the Center to several departments at HKU. As a first step toward a more complete integration with the Li Ka Shing Faculty of Medicine of HKU, we have targeted the establishment of joint appointments and have reached an agreement to nominate as Research Assistant Professor in the Department of Anatomy Dr Beatrice Nal-Rogier, who will be seconded to the Center. We have strengthened our team with the addition of another group led by Dr Hui-Ling YEN, who is holding an appointment as Research Assistant Professor in the Department of Microbiology. Dr Yen, who will move to the Center in August, has acquired an extensive training in molecular biology and influenza virology working as a PhD under the supervision of Professor Rob Webster (St. Jude’s Hospital, USA) and will continue her work on the role of specific genes from the highly pathogenic H5N1 virus as virulence factors in induction of host innate immune response. Her research project complements the interest of HKU-PRC and will contribute to develop our projects related to anti-viral immunity, thus compensating for the departure of Dr Joanna Ho, who left the center in September 2008. Dr Yen’s expertise in applying animal models and plasmid-based reverse genetics technique to study the biology of influenza virus will be a great asset to achieve our goals in research and teaching. This strategy of joint appointments, which we have discussed with Professor Sum-Ping Lee, the Dean of the Li Ka Shing Faculty of Medicine, will be pursued to facilitate the interactions of the Center with the Faculty, the career development of Junior Group Leaders and the gradual involvement of the Center in undergraduate teaching.

The Teaching and Training Center of Excellence in Biomedical Research has become operational and we now offer to HKU and international students an annual program with three postgraduate courses covering cell biology, virology and immunology. We have completed the renovation of the space on the 3rd floor of the Dexter HC Man building that was allocated to the Teaching and Training Center, thus enabling us to include practical sessions in the course programs. This has resulted in increased visibility and reputation of the center among the students, which has translated into an increased number of successful candidates applying for MPhil and PhD positions at HKU-PRC. This facility is also available to other departments and the Department of Biochemistry has contacted us to run the practical sessions of a “Stem Cell” course scheduled in August 2009.

We consider that these achievements validate the overall strategy presented to the Board in 2008 and constitute a solid foundation to ensure the continuous improvement of our performance, as well as the full participation of HKU-PRC in the academic life of HKU and Hong Kong. We envisage that the Center will be able to expand its scope and output by targeting an interdisciplinary approach and forging strategic partnerships with many departments, namely Anatomy, Microbiology, Pathology, Pediatrics and Adolescent Medicine, Pharmacology & Pharmacy, and Physiology as well as the AoE on influenza.

Research
HKU-PRC has currently 32 staff members:
17 scientific (8 students), 11 technical and 4 administrative. The scientific activity is broadly subdivided into three main areas (Virus-Host Interaction; Viral Infection & Immunity; Translational Research) that complement each other in both scientific goals and technical aspects.

The Virus-Host Interactions team led by Dr Beatrice Nal-Rogier has expanded its studies on the identification of novel cellular targets that interact with viral components during different phases of the viral life cycle. We have discovered distinct sets of genes that partner with structural proteins of influenza, dengue or human coronaviruses in a virus-specific manner. The roles of these factors in early and late stages of replication, as well as their implication for pathogenesis are under investigation. Two students will defend their PhD thesis at the end of 2009 by presenting their work on specific interactions between host and structural proteins of the SARS coronavirus. In particular, we have established an efficient protocol to produce Viral-Like Particles, which constitute a relevant and safe model to study molecular determinants of virion egress. The minimal requirement for assembly of VLPs for the coronavirus responsible for severe acute respiratory symdrome in humans (SARS-CoV) is still controversial. Our studies have uncovered the minimal viral protein set necessary for assembly of SARS coronavirus virions in host cells and revealed that the mechanism of SARS-CoV assembly differs from other studied coronaviruses (J Virol, 2008). As partners of the DENFRAME European consortium, an important achievement of the research program on dengue virus has been the development and molecular characterization of dengue virus like particles for the four virus serotypes. A Declaration of Invention has been filed to protect this technology with the aim of using dengue virus-like particles as a source of native antigens for antibody development and sero-diagnosis. The successful continuation of these projects has been secured by several external grants received from the Research Fund for the Control of Infectious Diseases, the Research Grant Council and the Area of Excellence on the “Control of Pandemic and Inter-pandemic Influenza”.

The Viral Infection & Immunity team led by Dr Martial Jaume focuses on the investigation of specific mechanisms underlying the interplay between viral infection and the immune response, innate and acquired by implementing research projects that are at the interface of fundamental and applied research. We are now exploring whether anti-influenza antibodies may perversely facilitate infection of humans by viruses from avian origin. Our preliminary results are highlighting a possible role of human heterosubtypic anti-influenza antibodies (i.e. anti-H1N1 and anti-H3N2) in triggering infection by avian H5 influenza virus. The efficient adaptation of an avian influenza virus to transmit from human-to-human raises fears worldwide about emergence of a deadly pandemic. Differential preference of binding of influenza viruses to sialylgalactosyl residues in humans versus birds is believed to restrict occurrence of zoonotic avian infection. However in addition to interaction with viral receptor, influenza viruses may also rely on anti-viral antibodies to enter and replicate into host cells, a phenomenon what has been demonstrated with H1N1 and H3N2 infections already. We have illustrated that immune-sera from individuals previously infected by H1 or H3 influenza viruses were not prominently efficient in neutralizing H5 viral infection. Conversely, those sera were triggering an enhanced infection of cells of the hematopoietic lineage when compared to controls (i.e. infection in absence of immune-serum). We are completing an RFCID grant centered on the role of innate immunity Natural Killer (NK) cells in the response to a challenge with the highly pathogenic avian influenza virus, which has already led to the publication of a manuscript (J Virol, 2008). We have submitted a new grant proposal to RFCID to continue this line of research which has recently been approved, pending minor revisions.

The activity of the Translational Research group, led by Dr Jean-Michel Garcia, is focused on the development of high throughput methodologies. The rationale for this option has been to create a strong interface between fundamental research and possible clinical applications. Three main axes of research have been followed: drug discovery, diagnostic and molecular biology. This diversity of application was derived from the flexibility of a single very potent tool: pseudotyped viral particles. The first drug screening campaign, validating the proof-of-principle of our cell-based assay has been published (Garcia et al., 2009) and other projects are being completed. We have carried out a third screening in collaboration with the Shanghai Institute of Materia Medica (SIMM), targeting inhibitors of dengue replication. This campaign has led to the identification of several leads and a Declaration of Invention has been filed. Furthermore, from lab-to-field, a novel sero-diagnostic assay for H5N1 has been transferred to partners in Cambodia and Vietnam and used in field investigations of H5N1 infection. We are further taking advantage of the properties of pseudotyped particles, which present the viral envelopes in their native conformation. They are referred to as chimerical virus like particles (VLP) when they are produced without any nucleic material encapsidated. These VLPs are intermediate between a virus and a liposome and are totally safe to handle. This technique presents a huge advantage when the original virus is a BSL-3 or BSL-4 agent and allows the use of Nuclear Magnetic Resonance (NMR) to investigate the interactions of the viral glycoprotein with host molecules at atomic level. This approach now opens up the possibility of a systematic analysis of the viral hemagglutinin-glycan receptor interactions of the highly pathogenic H5N1 virus. It provides, on the one hand, a novel tool to re-assess the glycans that bind the avian H5-haemagglutinin and, on the other hand, an exploration of the impact of viral mutations on such binding (Haselhorst et al, 2008). These projects have been funded through the Area of Excellence, the DENFRAME consortium, as well as grants-in–aid from the International Network of Pasteur Institutes.

Finally, although this Report deals chiefly with the achievements of the Center in 2008, it is important to place our activity also within the context of the recent outbreak of swine flu that has caused a great deal of alarm worldwide. Pandemic influenza remains the pre-eminent emerging infectious disease threat to global health and Hong Kong is located at the epicenter of a possible pandemic emergence. Being within Hong Kong but also being linked to the Pasteur Network of laboratories worldwide, and in Asia-Pacific in particular, we are well placed to be at the forefront of an early warning and response to emerging disease threats. In close collaboration with the department of Microbiology, HKU-PRC has contributed to the development of a novel, specific and sensitive test to detect the novel human H1N1 influenza virus. This test allows a rapid detection of the virus, can be performed with conventional PCR methods that are available in most laboratories in a BSL-2 containment and uses Asian swine H1N1 virus (A/SW/HK/PHK1578/03), which is not adapted to human transmission, and represents a much safer option as a positive control (Clin Chem, 2009). These reagents and methodology have already been supplied to a number of countries in the region.

Teaching & Education
The Teaching & Training Center of Excellence in Biomedical Research has been completed and we have launched two additional annual courses, HKU-Pasteur Immunology and HKU-Pasteur Cell Biology.
Both courses have been extremely well received and all the students were impressed by the quality of the program, of the lectures and of the invited speakers. In addition, we have been able to organize practical “hands-on” sessions for all courses, thus bringing an additional dimension to these Master Classes that challenge the students with concrete examples of experiment planning and interpretation of the results. All courses have been validated by HKU as part of its Research Postgraduate Curriculum and we are now training 30-40 HKU students every year. This teaching program is made possible through additional funds that we are receiving from the International Network of Pasteur Institutes, the Li Ka Shing Faculty of Medicine and the Croucher Foundation. Other sponsors (FCCIHK, The French Consulate and Sanofi-Pasteur) are also participating as indicated in the course reports. It is important to stress that such an annual program for top-quality courses is being realized for the first time in a center of the Pasteur network, with an important financial commitment from the International Network, and that HKU has responded enthusiastically to this mid-term approach by granting an annual contribution.

Our emphasis on cell imaging has led to the organization of two events in close collaboration with the Department of Anatomy: a Cell Imaging Workshop with special focus on “Fluorescence Lifetime Imaging Microscopy” in February 2008; and a Seminar Series in Cell Biology: Subcellular structures and Cellular Dynamics in November 2008, sponsored by the Croucher Foundation, with a panel of top local and international speakers. HKU-PRC continues its efforts to promote scientific exchanges between Hong Kong and Institut Pasteur through the Pasteur-Croucher Exchange Program. Two students are now working on their PhD thesis in Paris. We also plan on completing the integration of HKU-PRC into the department of Cell Biology and Infection of Institut Pasteur in order to become eligible to participate in the International PhD program launched by IP in 2009. Finally, HKU-PRC is committed to become involved in undergraduate teaching and, with the support of the Dean of the Medical School, we are making the appropriate contacts to implement this decision.

HKU-Pasteur Research Centre is firmly engaged in promoting knowledge exchange and showing community leadership. We have teamed up with the Victor Segalen French International School to organize a scientific workshop for grade 11th students. This project has been initiated by the Agency for French Teaching abroad, under the administration of the French Ministry of Foreign Affairs. The workshop consisted of two visits of the Centre for 2 groups of 17 students, during which they discovered various aspects of scientific research and work in an academic laboratory. A public conference at the School is also planned to conclude the program. It will present the values of research and of a scientific approach for discovering new pathogens. We also welcomed two students from the French International School for a one-week traineeship (March 2-6) during which they had the opportunity to follow different activities and be confronted with life in a research centre.

Another major achievement has been the contribution given by HKU-PRC in setting up the exhibition “From Plague to New Emerging Diseases: a Chronicle of Pasteurian Research in Hong Kong”, which was hosted by The University Museum and Art Gallery of HKU between May 27 and July 30, 2008. The exhibition explored the history of infectious diseases in Hong Kong from the plague of 1894 to the present day, and the challenges that virologists are still facing. The exhibition also showed the importance of global collaboration in the identification and prevention of deadly epidemics, such as dengue fever, avian influenza, and SARS. This event was a timely reminder of the lifesaving scientific medical collaboration between France and Hong Kong, and its implications for world health. Following this first display, some parts of the exhibition have been transferred to the Hong Kong Museum of Medical Sciences where it has been on display since mid-August. The resurgence of interest in Yersin’s contributions will culminate with the opening of a permanent exhibition on the “Discovery of the Cause of Plague”, sponsored by the French Consulate, and the unveiling of a bust of Alexandre Yersin, which will be installed in the Museum Garden.

HKU-PRC has collaborated with Dr Frederick Keck, a social anthropologist at the CNRS in Paris, who has joined us as a Visiting Scientist for the year 2008-2009. Dr Keck has been working on a project entitled “When animals make humans sick: A comparative anthropology of Avian Flu”, supported by a grant from the Fyssen Foundation. He is also a member of the French Centre for Research on Contemporary China, a cultural centre of the French Consulate in Hong Kong. Dr Keck has organized a major conference entitled “Avian Flu: Social and Anthropological Perspectives” co-sponsored by HKU-PRC and the Area of Excellence on “Control of pandemic and inter-pandemic influenza” (February 23-24, 2009). The goal of this conference was to discuss the social aspects of these networks and measures, and to open perspectives for the future at an international level. This conference has gathered microbiologists, social anthropologists and public health officials to study the impact of Avian Flu surveillance in different parts of the world (Asia, Europe, Africa, America) and at different stages of the spread of the virus (from animals to the humans). This conference has opened the way for an anthropological analysis of Avian Flu, integrating all the aspects of this global phenomenon: environmental, biological, social and psychological.

Network projects
HKU-PRC relies on a number of strategic collaborations and network projects. Dr Roberto Bruzzone is the Scientific Coordinator of the RESPARI and SISEA programs that federate the Institutes of the Pasteur-Asia network in a multi-center project. The RESPARI network emphasizes fundamental and translational research; it has established itself as one of the flagship research programs of the network having acquired international reputation, and has been instrumental in promoting active collaboration between the Pasteur network in Asia, HKU-PRC and HKU. It is worth mentioning here the close synergies with IP-Shanghai in the development of novel diagnostic tests and their validation (including recently during the swine flu crisis), the successful technology transfer achieved with IP-Cambodia, and the ground-breaking project with IP-Korea to carry out the first genome-wide screening in human cells to identify of host factors either restricting or facilitating influenza infection. More details are available on the RESPARI website (www.hku.hk.respari/). The aim of the Surveillance and Investigation of Endemic Situations in Southeast Asia (SISEA) program is to contribute to the improvement of the detection and handling of epidemic situations in the region. The SISEA project is a key initiative financed by the French Development Agency (AFD) (www.hku.hk/respari/research_07.htm). The specific goal of SISEA is to set up a surveillance and investigation program in four of the region’s countries (China, Laos, Vietnam, and Cambodia). As the Scientific Coordinator, Dr Bruzzone has carried out together with the Medical Advisor a thorough assessment of the progress made by the different partners and a great effort has been made to specifically address the major issues raised by the Scientific Advisory Board (SAB) during the 2007 meetings in Hong Kong and Shanghai. Moreover, important milestones have been achieved in the implementation and output of surveillance programs, which have put more emphasis on the expected patients’ benefit component. The coordination of this project significantly contributes to the recognition and visibility of HKU-PRC.

We have received the visit of Mrs Roselyne Bachelot-Narquin, French Minister of Health, Youth, Sport and the Voluntary Organizations, on August 9, 2008. We presented our mission, research and teaching activities and emphasized the role of Hong Kong as an influenza sentinel post. This is timely, as the management of large-scale safety risks such as influenza pandemic within the European Union was a major theme emphasized during a meeting of the European Union Health Ministers in September 2008. During her tour of the laboratories she discussed with staff scientists and students working on viral respiratory infections such as SARS and avian influenza. Madame Bachelot-Narquin praised the achievements of the Centre with the RESPARI program (partially financed by the French Ministry of Health) and reaffirmed the support of her Ministry.

Professor Malik Peiris is the Coordinator of the 8-year research program “Control of Pandemic and Inter-pandemic Influenza” that has been awarded a HK$ 76 millions by the University Grants Committee in the fourth round of its Areas of Excellence (AoE) scheme. The HKU-PRC takes an active part in this application and is involved in three work-packages: 1) To identify viral and host determinants of the pathogenesis of human H5N1 disease (WP2); and 2) To develop novel options for diagnosis, vaccines and therapy (WP5). We have secured substantial funds to successfully implement these specific research areas and strengthen our scientific collaborations with many groups at HKU and other universities in Hong Kong.

Professor Peiris also continues to serve on a number of WHO working groups in relation to both avian influenza H5N1 and more recently the swine origin influenza virus H1N1.

Financial situation
The financial situation has been defined under the Consolidated Agreement stipulating that the Centre will receive intramural funding from HKU up to the end of the revised term (November 2011). Following the 2008 Board Meeting, HKU has agreed to increase its annual support to HK$6.47 million in cash, keeping the existing level of in kind contributions). Starting in 2005 more intramural funds have been made available from Institut Pasteur, including a special donation to help the construction of a BSL-3 laboratory. Annual contributions from the University of Hong Kong and Institut Pasteur account for about 60% of current income. The remaining income is obtained through the competition for project-based external grants and donations. Over the past 3 years HKU-PRC has considerably increased its success rate in obtaining extramural grants from RGC (Research Grants Committee) and RFCID (Research Fund for the Control of Infectious Diseases) to support its core research activities. It is worth mentioning that in 2007 the Centre was managing only 1 RFCID grant, whereas we now hold 1 RGC, 3 RFCID and 3 Area of Excellence contracts, with 2 additional RFCID proposals approved pending minor modifications.

In summary, the financial position of the Centre is considered as healthy with a total net asset of HK$4.44M and a balance of cash and cash equivalent of HK$3.75M stood at June 30, 2008.