[Show abstract][Hide abstract] ABSTRACT: The chicken's major histocompatibility complex (MHC) haplotype has profound influence on the resistance or susceptibility to certain pathogens. For example, the B21 MHC haplotype confers resistance to Marek's disease (MD). However, non-MHC genes are also important in disease resistance. For example, lines 6 and 7 both express the B2 MHC haplotype, but differ in non-MHC genes. Line 6, but not line 7, is highly resistant to tumors induced by the Marek's disease herpesviruses and avian leukosis retroviruses. Recently, survival in the field by Thai indigenous chickens to H5N1 high-pathogenicity avian influenza (HPAI) outbreaks was attributed to the B21 MHC haplotype, whereas the B13 MHC haplotype was associated with high mortality in the field. To determine the influence of the MHC haplotype on HPAI resistance, a series of MHC congenic white leghorn chicken lines (B2, B12, B13, B19, and B21) and lines with different background genes but with the same B2 MHC haplotype (Line 63 and 7(1)) were intranasally challenged with low dose (10 mean chicken lethal doses) of reverse-genetics-derived rg-A/chicken/Indonesia/7/2003 (H5N1) HPAI virus. None of the lines were completely resistant to lethal effects of the challenge, as evidenced by mortality rates ranging from 40% to 100%. The B21 line had mortality of 40% and 70%, and the B13 line had mortality of 60% and 100% in two separate trials. In addition, the mean death times varied greatly between groups, ranging from 3.7 to 6.9 days, suggesting differences in pathogenesis. The data show that the MHC has some influence on resistance to AI, but less than previously proposed, and non-MHC background genes may have a bigger influence on resistance than the MHC.
[Show abstract][Hide abstract] ABSTRACT: Vaccination against avian influenza (AI) virus, a powerful tool for control of the disease, may result in issues related to surveillance programs and international trade of poultry and poultry products. The use of AI vaccination in poultry would have greater worldwide acceptance if a reliable test were available that clearly discriminated between naturally infected and vaccinated-only animals (DIVA). Because the nonstructural protein (NS1) is expressed in influenza virus-infected cells, and it is not packaged in the virion, it is an attractive candidate for a DIVA differential diagnostic test. The aim of this work was to determine the onset of the antibody response to the NS1 protein in chickens infected with low pathogenic avian influenza (LPAI) virus, and to evaluate the diagnostic potential of a baculovirus-expressed purified NS1 protein in an indirect ELISA-based DIVA strategy. An antibody response against NS1 was first detected 3 wk after infection, but the antibody levels were decreasing rapidly by 5 wk after infection. However, most chickens did not have detectable antibodies in spite of high hemagglutination inhibition (HI) antibody titers in one group. In birds vaccinated with inactivated oil-emulsion vaccines, antibodies against NS1 were not detected before virulent challenge, and only a small percentage of birds seroconverted after homologous LPAI virus challenge. Vaccinated birds challenged with highly pathogenic AI showed a higher NS1 antibody response, but at most only 40% of birds seroconverted against NS1 protein by 3 wk after challenge. Because of the variability of seroconversion and the duration of the antibody response in chickens, the NS1 protein DIVA strategy did not perform as well as expected, and if this strategy were to be used, it would require sampling a higher number of birds to compensate for the lower seroconversion rate.
[Show abstract][Hide abstract] ABSTRACT: The first known cases of human infection with highly pathogenic avian influenza (HPAI) H5N1 viruses in Vietnam occurred in late 2003. However, HPAI H5N1 and low-pathogenic avian influenza (LPAI) H5N2 and H9N3 viruses were isolated from domestic waterfowl during live-bird market (LBM) surveillance in Vietnam in 2001 and 2003. To understand the possible role of these early viruses in the genesis of H5N1 strains infecting people, we performed sequencing and molecular characterization. Phylogenetic analysis revealed that the hemagglutinin (HA) genes of two geese HPAI H5N1 strains belonged to clade 3, and their surface glycoprotein and replication complex genes were most closely related (98.5-99.7% homologous) to A/duck/Guangxi/22/01 (H5N1) virus, detected contemporarily in southern China, whilst the M and NS genes were derived from an A/duck/Hong Kong/2986.1/00 (H5N1)-like virus. The H5 HA gene of the duck HPAI H5N1 strain belonged to clade 5 and acquired a gene constellation from A/quail/Shantou/3846/02 (H5N1), A/teal/China/2978.1/02 (H5N1) and A/partridge/Shantou/2286/03 (H5N1)-like viruses. The phylogenetic analysis further indicated that all eight gene segments of goose and duck HPAI H5N1 and LPAI H5N2 viruses were distinct from those of H5N1 clade-1 viruses known to have caused fatal human infections in Vietnam since late 2003. The duck H9N3 isolates derived genes from aquatic-bird influenza viruses, and their H9 HA belonged to the Korean lineage. The PB2 gene of A/duck/Vietnam/340/01 (H9N3) virus had lysine at position 627. Based on the molecular characterization of specific amino acid residues in the surface and relevant internal protein-coding genes, the Vietnamese H5N1 and H9N3 virus isolates indicated specificity to avian cell surface receptor and susceptibility for currently licensed anti-influenza A virus chemotherapeutics. Our findings suggest that the H5N1 and H5N2 viruses that circulated among geese and ducks in LBMs in Hanoi, Vietnam, during 2001 and 2003 were not the immediate ancestors of the clade-1 viruses associated with fatal human infections in Vietnam. The clade-1 HPAI H5N1 viruses were independently introduced into Vietnam.
Archives of Virology 08/2009; 154(8):1249-61. DOI:10.1007/s00705-009-0429-2 · 2.39 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Highly pathogenic avian influenza (HPAI) H5N1 viruses have caused dramatic economic losses to the poultry industry of Vietnam and continue to pose a serious threat to public health. As of June 2008, Vietnam had reported nearly one third of worldwide laboratory confirmed human H5N1 infections. To better understand the emergence, spread and evolution of H5N1 in Vietnam we studied over 300 H5N1 avian influenza viruses isolated from Vietnam since their first detection in 2001. Our phylogenetic analyses indicated that six genetically distinct H5N1 viruses were introduced into Vietnam during the past seven years. The H5N1 lineage that evolved following the introduction in 2003 of the A/duck/Hong Kong/821/2002-like viruses, with clade 1 hemagglutinin (HA), continued to predominate in southern Vietnam as of May 2007. A virus with a clade 2.3.4 HA newly introduced into northern Vietnam in 2007, reassorted with pre-existing clade 1 viruses, resulting in the emergence of novel genotypes with neuraminidase (NA) and/or internal gene segments from clade 1 viruses. A total of nine distinct genotypes have been present in Vietnam since 2001, including five that were circulating in 2007. At least four of these genotypes appear to have originated in Vietnam and represent novel H5N1 viruses not reported elsewhere. Geographic and temporal analyses of H5N1 infection dynamics in poultry suggest that the majority of viruses containing new genes were first detected in northern Vietnam and subsequently spread to southern Vietnam after reassorting with pre-existing local viruses in northern Vietnam. Although the routes of entry and spread of H5N1 in Vietnam remain speculative, enhanced poultry import controls and virologic surveillance efforts may help curb the entry and spread of new HPAI viral genes.
PLoS ONE 02/2008; 3(10):e3462. DOI:10.1371/journal.pone.0003462 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: An outbreak of highly pathogenic avian influenza A
(H5N1) has recently spread to poultry in 9 Asian countries.
H5N1 infections have caused >52 human deaths in
Vietnam, Thailand, and Cambodia from January 2004 to
April 2005. Genomic analyses of H5N1 isolates from birds
and humans showed 2 distinct clades with a nonoverlapping
geographic distribution. All the viral genes were of avian
influenza origin, which indicates absence of reassortment
with human influenza viruses. All human H5N1 isolates tested
belonged to a single clade and were resistant to the
adamantane drugs but sensitive to neuraminidase
inhibitors. Most H5N1 isolates from humans were antigenically
homogeneous and distinct from avian viruses circulating
before the end of 2003. Some 2005 isolates showed
evidence of antigenic drift. An updated nonpathogenic H5N1
reference virus, lacking the polybasic cleavage site in the
hemagglutinin gene, was produced by reverse genetics in
anticipation of the possible need to vaccinate humans.
[Show abstract][Hide abstract] ABSTRACT: Since 1997, outbreaks of highly pathogenic (HP) H5N1 and circulation of H9N2 viruses among domestic poultry in Asia have posed a threat to public health. To better understand the extent of transmission of avian influenza viruses (AIV) to humans in Asia, we conducted a cross-sectional virologic study in live bird markets (LBM) in Hanoi, Vietnam, in October 2001. Specimens from 189 birds and 18 environmental samples were collected at 10 LBM. Four influenza A viruses of the H4N6 (n = 1), H5N2 (n = 1), and H9N3 (n = 2) subtypes were isolated from healthy ducks for an isolation frequency of over 30% from this species. Two H5N1 viruses were isolated from healthy geese. The hemagglutinin (HA) genes of these H5N1 viruses possessed multiple basic amino acid motifs at the cleavage site, were HP for experimentally infected chickens, and were thus characterized as HP AIV. These HA genes shared high amino acid identities with genes of other H5N1 viruses isolated in Asia during this period, but they were genetically distinct from those of H5N1 viruses isolated from poultry and humans in Vietnam during the early 2004 outbreaks. These viruses were not highly virulent for experimentally infected ducks, mice, or ferrets. These results establish that HP H5N1 viruses with properties similar to viruses isolated in Hong Kong and mainland China circulated in Vietnam as early as 2001, suggest a common source for H5N1 viruses circulating in these Asian countries, and provide a framework to better understand the recent widespread emergence of HP H5N1 viruses in Asia.
Journal of Virology 05/2005; 79(7):4201-12. DOI:10.1128/JVI.79.7.4201-4212.2005 · 4.44 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Pandemic vaccine seed viruses for H5N1, H5N2 and H9N2 subtypes were prepared and evaluated. Formalin-inactivated reassortant vaccines using A/Puerto Rico/8/34 (PR8) as a donor of internal genes with surface glycoprotein genes, hemagglutinin (HA) and neuraminidase (NA), of H5N2 and H9N2 subtypes, were generated by conventional reassortment. A reassortant virus for the highly pathogenic H5N1 subtype was prepared using reverse genetics with a genetically modified HA gene. All candidate vaccines were immunogenic in mice and ferrets, and mice immunized with inactivated vaccines were completely protected against challenge infection with wild-type parent viruses. All viruses were non-pathogenic for chickens in the standard USDA pathotyping test.
International Congress Series 06/2004; 1263:813-817. DOI:10.1016/j.ics.2004.01.045