HKU-PASTEUR RESEARCH CENTRE LIMITED
| Researcher : Altmeyer RM |
| List of Research Outputs |
| Wang P., Kudelko M., Lo J.C.Y., Siu Y.L., Kwok K.T.H., Sachse M., Nicholls J.M., Bruzzone R., Altmeyer R.M. and Nal-Rogier B.T.M., Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins, PLoS ONE. 2009, 4(12): e8325. |
| Researcher : Bruzzone R |
| Project Title: | Regulation of early steps of coronavirus infection by the ezrin ERM protein |
| Investigator(s): | Bruzzone R, Cheung CY, Nal-Rogier BTM |
| Department: | Medical Faculty |
| Source(s) of Funding: | General Research Fund (GRF) |
| Start Date: | 11/2008 |
| Abstract: |
| (1) To identify the determinants of interaction between the SARS-CoV Spike carboxy-terminal domain and cellular ezrin proteins - Definition of the amino-acids of Spike CT involved in ezrin binding by mutagenesis and GST pull-down assay - Differential binding of ezrin by SARS-CoV Spike versus other human coronavirus Spike (group 1: 229E, NL63; group 2: HKUI, OC43) - Synthesis of small peptides that mimic the Spike ezrin-binding motif and test for inhibition of Spike-ezrin interaction in GST-pull down experiments - Analysis of ezrin subcellular localization/recruitment by SARS-CoV Spike wild-type and mutant forms as well as other human coronavirus Spikes in transfected cells by immuno-fluorescence studies; (2) To test the role of ezrin in SARS-CoV virus infection - Measurement of SARS-CoV infection in susceptible FrHK4 cells where ezrin would have been either knocked-down by siRNA targeting or over-expressed (experiment performed in BSL3 laboratory). Both viral RNAs (positive and negative strands for N and ORF1ab genes) levels and virion production will be measured by quantitative RT-PCR and infectivity assays, respectively. - Effect of inhibitory peptides will be investigated on SARS-CoV life cycle; (3) To define the precise role of SARS-CoV Spike CT / ezrin interaction in the entry process of SARS-CoV - Production of lentiviral particles pseudotyped with wild-type and mutant forms of the SARS-CoV Spike protein (SARSpp and mSARSpp) - Infectivity assays using SARSpp and mSARSpp - Effect of inhibitory peptides on SARSpp infectivity - Analysis of ezrin recruitment at the site of SARS-CoV entry in infected cells by fluorescence microscopy. |
| List of Research Outputs |
| Bruzzone R., Infectious Diseases of the Nervous System and Their Impact in Developing Countries, In: Roberto Bruzzone, Monique Dubois-Dalcq, Georges E. Grau, Diane E. Griffin, Krister Kristensson, PLoS Pathogens. 2010, 5: e1000199. |
| Wang P., Kudelko M., Lo J.C.Y., Siu Y.L., Kwok K.T.H., Sachse M., Nicholls J.M., Bruzzone R., Altmeyer R.M. and Nal-Rogier B.T.M., Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins, PLoS ONE. 2009, 4(12): e8325. |
| Researcher : Garcia J |
| List of Research Outputs |
| Buchy P., Vong S., Garcia J., Chu S., Hien T.T., Hien V.M., Channa M., Ha D.Q., Vinh Chau N.V., Simons C., Farrar J.J., Peiris J.S.M. and de Jong M.D., Kinetics of neutralizing antibodies in patients naturally infected by H5N1 virus. , PlosOne. 2010, 5: e10864. |
| Cavailler P., Chu S., Ly S., Garcia J., Ha D.Q., Bergeri I., Som L., Ly S., Sok T., Vong S. and Buchy P., Seroprevalence of anti-H5 antibodies in rural Cambodia, 2007, Journal of clinical virology. 2010, 48: 123-6. |
| Garcia J., Pepin S., Lagarde N.J.C., Ma S.K., vogel F.R., Chan K.H., Chiu S.S.S. and Peiris J.S.M., Heterosubtype neutralizing responses to influenza A (H5N1) viruses are mediated by antibodies to virus haemagglutinin, PLoS One. 2009, 4(11): e7918. |
| Garcia J., Fries K., Lagarde N.J.C., Ma S.K., Buchy P., de Jong M.D. and Peiris J.S.M., Optimization and evaluation of an influenza A (H5) pseudotyped lentiviral particle – based serological assay. , Journal of clinical virology. 2010, 47: 29-33. |
| Lai J.C.C., Chan W.W.L., Kien F.S., Nicholls J.M., Peiris J.S.M. and Garcia J., Formation of virus-like particles from human cell lines exclusively expressing Influenza neuraminidase, Journal of General Virology . 2010, 2322-30. |
| Schultsz C., Dung N.V., Hai L.T., Ha D.Q., Peiris J.S.M., Lim W., Garcia J., Tho N.D., Lan H., Thao P.X., Rogier van Doorn H., Vinh Chau N.V., Farrar J. and de Jong M.D., Prevalence of antibodies against avian Influenza A (H5N1) virus among cullers and poultry workers in Ho Chi Minh City, 2005. , PlosOne. 2009, 4: e7948. |
| Researcher : Kien FS |
| List of Research Outputs |
| Lai J.C.C., Chan W.W.L., Kien F.S., Nicholls J.M., Peiris J.S.M. and Garcia J., Formation of virus-like particles from human cell lines exclusively expressing Influenza neuraminidase, Journal of General Virology . 2010, 2322-30. |
| Researcher : Kudelko M |
| List of Research Outputs |
| Wang P., Kudelko M., Lo J.C.Y., Siu Y.L., Kwok K.T.H., Sachse M., Nicholls J.M., Bruzzone R., Altmeyer R.M. and Nal-Rogier B.T.M., Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins, PLoS ONE. 2009, 4(12): e8325. |
| Researcher : Kwok KTH |
| List of Research Outputs |
| Wang P., Kudelko M., Lo J.C.Y., Siu Y.L., Kwok K.T.H., Sachse M., Nicholls J.M., Bruzzone R., Altmeyer R.M. and Nal-Rogier B.T.M., Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins, PLoS ONE. 2009, 4(12): e8325. |
| Researcher : Lagarde NJC |
| List of Research Outputs |
| Garcia J., Pepin S., Lagarde N.J.C., Ma S.K., vogel F.R., Chan K.H., Chiu S.S.S. and Peiris J.S.M., Heterosubtype neutralizing responses to influenza A (H5N1) viruses are mediated by antibodies to virus haemagglutinin, PLoS One. 2009, 4(11): e7918. |
| Garcia J., Fries K., Lagarde N.J.C., Ma S.K., Buchy P., de Jong M.D. and Peiris J.S.M., Optimization and evaluation of an influenza A (H5) pseudotyped lentiviral particle – based serological assay. , Journal of clinical virology. 2010, 47: 29-33. |
| Researcher : Lai JCC |
| List of Research Outputs |
| Lai J.C.C., Chan W.W.L., Kien F.S., Nicholls J.M., Peiris J.S.M. and Garcia J., Formation of virus-like particles from human cell lines exclusively expressing Influenza neuraminidase, Journal of General Virology . 2010, 2322-30. |
| Researcher : Nal-Rogier BTM |
| List of Research Outputs |
| Wang P., Kudelko M., Lo J.C.Y., Siu Y.L., Kwok K.T.H., Sachse M., Nicholls J.M., Bruzzone R., Altmeyer R.M. and Nal-Rogier B.T.M., Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins, PLoS ONE. 2009, 4(12): e8325. |
| Researcher : Peiris JSM |
| Project Title: | IV International Symposium on Respiratory Viral Infections The Influenza Disease Burden in Hong Kong: the Rates of Hospitalisation in Children |
| Investigator(s): | Peiris JSM |
| Department: | Microbiology |
| Source(s) of Funding: | URC/CRCG - Conference Grants for Teaching Staff |
| Start Date: | 11/2001 |
| Abstract: |
| N/A |
| Project Title: | Influenza pandemic preparedness in Asia (Research collaboration with national Institute of Allergy and Infectious Diseases (NIAID), St. Jude Children's Research Hospital, Inc. (SJCRH) and The University of Hong Kong) |
| Investigator(s): | Peiris JSM, Guan Y |
| Department: | Microbiology |
| Source(s) of Funding: | National Institutes of Health, US Department of Health and Human Services - General Award |
| Start Date: | 04/2003 |
| Abstract: |
| To study influenza pandemic preparedness in Asia. |
| Project Title: | Virus-cell interaction and the pathogenesis of SARS |
| Investigator(s): | Peiris JSM |
| Department: | Microbiology |
| Source(s) of Funding: | VCO SARS Research Fund |
| Start Date: | 07/2003 |
| Abstract: |
| To explore what is the biological basis for the selective attraction of macrophages to the lung; to study what immunopathological consequences follow. |
| Project Title: | Viral determinants underlying the increased secretion of pro-inflammatory cytokines in H5N1 infected human macrophages: a mechanism for the unusual severity of human H5N1 disease? |
| Investigator(s): | Peiris JSM, Guan Y, Lau ASY, Poon LLM |
| Department: | Microbiology |
| Source(s) of Funding: | General Research Fund (GRF) |
| Start Date: | 09/2003 |
| Abstract: |
| To identify the viral genetic determinants responsible for the differential cytokine hyper-induction associated with H5N1/97 viruses. We have already established the role of the viral NS gene, however, a role for other viral genes has not been excluded. we hypothesize (and have preliminary evidence) that one or more other viral genes are also involved in the hyper-induction of cytokines; to determine whether the avian viral genes/gene products of H5N1/97 directly up-regulate cytokine induction or lack "immune evasion" mechanisms evolved by human influenza viruses? to determine whether the differential up-regulation of cytokines associated with H5N1/97 is also observed in human lung epithelial cells (primary bronchial epithelial cells and human lung epithelial cell lines e.g. A549 cells. |
| Project Title: | The macrophage in the pathogenesis of Severe Acute Respiratory Syndrome Coronavirus infection |
| Investigator(s): | Peiris JSM |
| Department: | Microbiology |
| Source(s) of Funding: | Research Fund for the Control of Infectious Diseases - Full Grants |
| Start Date: | 12/2005 |
| Abstract: |
| The global gene expression profiling of SARS-CoV infected macrophages will provide the basis for a more detailed analysis of the signaling pathways involved in subsequent studies, and may provide rational and novel therapeutic options to be devised for treating SARS. |
| Project Title: | Mass Tag PCR: A novel, multiplex and potentially sensitive approach for the rapid detection of respiratory pathogens |
| Investigator(s): | Peiris JSM, Chiu SSS, Cheung CY |
| Department: | Microbiology |
| Source(s) of Funding: | Research Fund for the Control of Infectious Diseases - Full Grants |
| Start Date: | 06/2006 |
| Abstract: |
| To divise, optimize and evaluate the Mass Tag PCR system for the rapid disgnosis for simultaneous screening of a wide spectrum of causative agents for human respiratory tract infections, including the avian influenza H5N1 and H7N7, SARS-coronavirus (SARS-CoV). Thiw will significantly facilitate better prevention, treatment and control of infectious diseases. |
| Project Title: | The pathogenesis of avian influenza A H5N1 in humans: macrophage and epithelial cell-virus interactions in vitro and ex-vivo |
| Investigator(s): | Peiris JSM |
| Department: | Microbiology |
| Source(s) of Funding: | The Wellcome Trust - General Award |
| Start Date: | 01/2007 |
| Abstract: |
| Specific aims are to: 1) Characterise the influenza viral replication profiles in tissue explant (ex-vivo) cultures of nasopharyngeal biopsies and lung tissue and primary cell cultures of human macrophages and respiratory (nasopharyngeal, bronchial, alveolar) epithelium. 2) Charaterise the functional receptors for H5N1 virus on macrophages and nasopharyngeal epithelium. 3) Define the consequences of viral entry via alternative pathways including the entry of non-neutralised virus antibody complexes via the Fc and complement receptors, to determine how these pathways affects subsequent viral replication and /or innate immune responses such as cytokine/chemokine secretion. |
| Project Title: | Communicable Diseases Surveillance and Response (CSR) - Standardization and shipment of PCR diagnostic kit for Avian Influenza detection to Member countries (viz: Bangladesh, Indonesia, Myanmar, Sri Lanka and Thailand) |
| Investigator(s): | Peiris JSM |
| Department: | Microbiology |
| Source(s) of Funding: | Collaborating Project Fund |
| Start Date: | 06/2007 |
| Abstract: |
| To compile diagnostic reagents in the form of kits for RT-PCR detection of H5N1 influenza for the South East Asia Region countries on behalf of WHO / SEAR. |
| Project Title: | Control of Pandemic and Inter-pandemic Influenza |
| Investigator(s): | Peiris JSM, Guan Y, Yuen KY, Chen H, Leung GM, Lau YL |
| Department: | Microbiology |
| Source(s) of Funding: | Areas of Excellence Scheme |
| Start Date: | 01/2008 |
| Abstract: |
| nil |
| Project Title: | Effector roles of cytokine cascades in the pathogenesis of human H5N1 disease |
| Investigator(s): | Peiris JSM, Cheung CY, Lee MY, Nicholls JM |
| Department: | Microbiology |
| Source(s) of Funding: | General Research Fund (GRF) |
| Start Date: | 01/2008 |
| Abstract: |
| To investigate the effect of soluble mediators from H5N1 infected cells (macrophages, lung epithelium) in relation to the activation of secondary cytokines (viz. the secondary cytokine cascades), and the gene expression related to apoptosis, oxidative stress, nitric oxide and other stress / toxicity pathways; to investigate the effect of such soluble mediators in relation to the induction of an antiviral state. |
| List of Research Outputs |
| Chan K.H., Lai S.T., Poon L.L.M., Guan Y., Yuen K.Y., Peiris J.S.M. and Peiris J.S.M., Analytical sensitivity of rapid influenza antigen detection tests for swine-origin influenza virus (H1N1). , J Clin Virol. . 2009, 45: 205-7. |
| Chiu S.S.S., Chan K.H., Tu W., Lau Y.L. and Peiris J.S.M., Immunogenicity and safety of intradermal versus intramuscular route of influenza immunization in infants less than 6 months of age: a randomized controlled trial, Vaccine. 2009, 27(35): 4834-9. |
| Garcia J., Pepin S., Lagarde N.J.C., Ma S.K., vogel F.R., Chan K.H., Chiu S.S.S. and Peiris J.S.M., Heterosubtype neutralizing responses to influenza A (H5N1) viruses are mediated by antibodies to virus haemagglutinin, PLoS One. 2009, 4(11): e7918. |
| Garcia J., Fries K., Lagarde N.J.C., Ma S.K., Buchy P., de Jong M.D. and Peiris J.S.M., Optimization and evaluation of an influenza A (H5) pseudotyped lentiviral particle – based serological assay. , Journal of clinical virology. 2010, 47: 29-33. |
| Mao H., Tu W., Liu Y., Qin G., Zheng J., Chan P.L., Lam K.T., Peiris J.S.M. and Lau Y.L., Inhibition of human natural killer cell activity by influenza virions and hemagglutinin, J Virol. 2010, 84(9): 4148-57. |
| Peiris J.S.M., Tu W., Yen H., Peiris J.S.M. and Yen H., A novel H1N1 virus causes the first pandemic of the 21st century, Eur J Immunol. 2009, 39(11): 2946-54. |
| Peiris J.S.M., Hui P.Y., Yen H., Peiris J.S.M. and Yen H., Host response to influenza virus: protection versus immunopathology, In: Adolfo Garcia-Sastre and Philippe Sansonetti, Current Opinion in Immunology. 2010, 22: 475-481. |
| Qin G., Mao H., Zheng J., Sia S.F., Liu Y., Chan P.L., Lam K.T., Peiris J.S.M., Lau Y.L. and Tu W., Phosphoantigen-expanded human gammadelta T cells display potent cytotoxicity against monocyte-derived macrophages infected with human and avian influenza viruses, J Infect Dis. 2009, 200(6): 858-65. |
| Schultsz C., Dung N.V., Hai L.T., Ha D.Q., Peiris J.S.M., Lim W., Garcia J., Tho N.D., Lan H., Thao P.X., Rogier van Doorn H., Vinh Chau N.V., Farrar J. and de Jong M.D., Prevalence of antibodies against avian Influenza A (H5N1) virus among cullers and poultry workers in Ho Chi Minh City, 2005. , PlosOne. 2009, 4: e7948. |
| Researcher : Siu YL |
| List of Research Outputs |
| Wang P., Kudelko M., Lo J.C.Y., Siu Y.L., Kwok K.T.H., Sachse M., Nicholls J.M., Bruzzone R., Altmeyer R.M. and Nal-Rogier B.T.M., Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins, PLoS ONE. 2009, 4(12): e8325. |
| Researcher : Tang D |
| List of Research Outputs |
| Tang D., Dong S., Ma N.F., Xie D., Chen L., Fu L., Lau S.H., Li Y., Li Y. and Guan X.Y., Overexpression of eukaryotic initiation factor 5A2 enhances cell motility and promotes tumor metastasis in hepatocellular carcinoma, Hepatology. 2010, 51: 1255-63. |
| Researcher : Wang P |
| List of Research Outputs |
| Wang P., Kudelko M., Lo J.C.Y., Siu Y.L., Kwok K.T.H., Sachse M., Nicholls J.M., Bruzzone R., Altmeyer R.M. and Nal-Rogier B.T.M., Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins, PLoS ONE. 2009, 4(12): e8325. |
| Researcher : Yen H |
| Project Title: | Identification of influenza neuraminidase sialyl substrates using glycan arrays |
| Investigator(s): | Yen H, Peiris JSM, Nicholls JM |
| Department: | Microbiology |
| Source(s) of Funding: | Seed Funding Programme for Basic Research |
| Start Date: | 05/2009 |
| Completion Date: | 05/2010 |
| Abstract: |
| The two surface glycoproteins of influenza A and B viruses both recognize sialic acid but with counteracting functions: hemagglutinin (HA) binds to sialic acid-containing receptors and neuraminidase (NA) hydrolyzes α-ketosidic linkage between sialic acid and the adjacent oligosaccharide (27). Sialic acid (N-acetylneuraminic acid [NeuAc], N-glycolylneuramic acid [NeuGc], and deaminoneuraminic acid [KDN]) is ubiquitously expressed on the cells surface of vertebrates, and the terminal sialic acid residues are usually linked to the adjacent oligosaccharide (galactose [Gal], N-acetylgalactosamine [GalNAc], N-acetylglucosamine [GlcNAc], or another sialic acid) via an α2,3-, α2,6- , α2,8-, or α2,9- linkage(7). For influenza viruses, selection of target cells is mediated by the HA receptor binding domain (21), although the role of NA cannot be excluded (14). The HA of Influenza viruses specifically recognize NeuAc and NeuGc sialic acid species, and the linkage connecting sialic acid and the adjacent sugar is important for determining the host range of influenza A viruses, as the avian and human influenza viruses were shown to preferentially recognize sialic acid linked to adjacent Gal via α2,3- or α2,6- linkages, respectively (19,20). In addition to the terminal sialic acid linkages, glycan array screening has identified that internal linkages as well as fucosylation, sulfation, and sialylation at the inner oligosaccharide also determined HA receptor recognition (13,23,24). A balanced HA and NA function has been shown to determine virus replication efficiency (11,27). As the HA adapts in different hosts, it is expected that the NA substrate specificity may alter correspondingly. While much interest is focused on identifying glycans recognized by the HA, it is also important to evaluate the spectrum of sialyl substrates processed by NA. Although previous studies have shown that NA from human and avian influenza virus differed in processing α2,3- or α2,6- linked sialyl receptors, only limited α2,3- or α2,6- linked synthesized glycans of simple structures were included in the studies (4,10,12,15). Due to the variety of the sialyl glycans presented in vivo, a large scale screening for potential NA substrate is needed. We first hypothesize that in addition to the terminal sialic acid linkages, the internal linkages as well as modifications of the inner oligosaccharides can affect NA substrate specificity. We further hypothesize that as HA adapts in different hosts, differences in NA specificity and activity exist between adapted influenza viruses. These hypotheses are based on the following observations. First, study of seven bacterial NAs revealed that differences in sialidase activity depend on the terminal sialic acid structure, terminal sialic acid linkages, and the structure of the adjacent sugar (7). Second, study of six influenza H1N1 viruses from different host origins revealed differences in substrate specificity as well as hydrolytic activity. While all six NAs have a higher activity for α2,3- sialosaccharides, the duck virus showed 50 times higher efficiency for α2,3- than α2,6- sialosaccharides and the human virus showed comparable efficiency in hydrolyzing α2,3- and α2,6- sialosaccharide (16). Third, a balanced HA and NA activity is needed for efficient viral growth. As the H2 and H3 that originated from an avian origin gradually adapted in human host, a change in N2 NA substrate specificity has been observed (5). The molecular determinants of NA substrate specificity was further mapped in proximity of the NA active site (12). However, the ligands for HA and NA may not always match as shown by recent human H3N2 isolates (8). Based on these studies, the specific aims of this proposal are first to apply non-infectious lentivirus-like particles (VLPs) to identify NA substrates using glycan arrays followed by determination of enzyme kinetics with the identified glycans. The benefit of using VLPs is that they retain the biological functions of surface glycoproteins, and the HA and NA expression level on a VLP should resemble that of an intact virus. Further, applying the VLP system, we can study the role of HA or NA alone or evaluate their function at the same time. We aim to focus on the N1 subtype for the preliminary experiments with long term goals of extending the study to N2 subtype as well as studying the interaction between HA and NA of a given strain. SPECIFIC AIMS: 1. Focusing on the N1 subtype, we will apply VLPs to identify the sialyl substrates of human influenza viruses, A/Hong Kong/218847/06 (H1N1) and A/Vietnam/1203/04 (H5N1), as well as avian influenza viruses, A/Duck/Alberta/35/76 (H1N1) on printed covalently linked glycan arrays. 2. Determine NA enzyme kinetics with identified glycans in a 96-well setting. |
| Project Title: | The role of balanced hemagglutinin-neuraminidase activity on the genesis of transmissible NA inhibitor-resistant variants in seasonal and novel pandemic influenza A H1N1 viruses |
| Investigator(s): | Yen H, Peiris JSM |
| Department: | Microbiology |
| Source(s) of Funding: | Research Fund for the Control of Infectious Diseases - Full Grants |
| Start Date: | 01/2010 |
| Abstract: |
| To assess the transmission potential of NA inhibitor-resistant pandemic H1N1 2009 viruses. |
| List of Research Outputs |
| Peiris J.S.M., Tu W., Yen H., Peiris J.S.M. and Yen H., A novel H1N1 virus causes the first pandemic of the 21st century, Eur J Immunol. 2009, 39(11): 2946-54. |
| Peiris J.S.M., Hui P.Y., Yen H., Peiris J.S.M. and Yen H., Host response to influenza virus: protection versus immunopathology, In: Adolfo Garcia-Sastre and Philippe Sansonetti, Current Opinion in Immunology. 2010, 22: 475-481. |