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1.
Emerg Infect Dis ; 23(4): 691-695, 2017 04.
Article in English | MEDLINE | ID: mdl-28322695

ABSTRACT

Highly pathogenic avian influenza viruses (HPAIVs) A(H5N6) were concurrently introduced into several distant regions of Japan in November 2016. These viruses were classified into the genetic clade 2.3.4.4c and were genetically closely related to H5N6 HPAIVs recently isolated in South Korea and China. In addition, these HPAIVs showed further antigenic drift.


Subject(s)
Influenza A virus/genetics , Influenza A virus/pathogenicity , Influenza in Birds/virology , Animals , Birds , Influenza in Birds/epidemiology , Influenza in Birds/mortality , Japan , Phylogeny
2.
Virus Genes ; 53(1): 44-51, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27738904

ABSTRACT

Since 2014, clade 2.3.4.4 H5 subtype highly pathogenic avian influenza viruses (HPAIVs) have been distributed worldwide. These viruses, which were reported to be highly virulent in chickens by intravenous inoculation, have a consensus HPAI motif PLRERRRKR at the HA cleavage site. However, two-clade 2.3.4.4 H5N8 viruses which we isolated from wild migratory birds in late 2014 in Japan possessed atypical HA cleavage sequences. A swan isolate, Tottori/C6, had a novel polybasic cleavage sequence, PLGERRRKR, and another isolate from a dead mandarin duck, Gifu/01, had a heterogeneous mixture of consensus PLRERRRKR and variant PLRERRRRKR sequences. The polybasic HA cleavage site is the prime virulence determinant of AIVs. Therefore, in the present study, we examined the pathogenicity of these H5N8 isolates in chickens by intravenous inoculation. When 106 EID50 of these viruses were intravenously inoculated into chickens, the mean death time associated with Tottori/C6 was substantially longer (>6.1 days) than that associated with Gifu/01 (2.5 days). These viruses had comparable abilities to replicate in tissue culture cells in the presence and absence of exogenous trypsin, but the growth of Tottori/C6 was hampered. These results indicate that the novel cleavage motif of Tottori/C6 did not directly affect the infectivity of the virus, but Tottori/C6 caused attenuated pathogenicity in chickens because of hampered replication efficiency. It is important to test for the emergence of diversified HPAIVs, because introduction of HPAIVs with a lower virulence like Tottori/C6 might hinder early detection of affected birds in poultry farms.


Subject(s)
Amino Acid Motifs , Hemagglutinin Glycoproteins, Influenza Virus/chemistry , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Influenza A Virus, H5N8 Subtype/genetics , Influenza in Birds/virology , Amino Acid Sequence , Animals , Animals, Wild/virology , Cell Line , Chickens , Hemagglutinin Glycoproteins, Influenza Virus/metabolism , Influenza A Virus, H5N8 Subtype/metabolism , Influenza A Virus, H5N8 Subtype/pathogenicity , Influenza in Birds/mortality , Phylogeny , Poultry Diseases/virology , Sequence Analysis, DNA , Viral Load , Virulence , Virus Replication
3.
J Gen Virol ; 97(9): 2104-2116, 2016 09.
Article in English | MEDLINE | ID: mdl-27260213

ABSTRACT

The nucleoprotein (NP) possesses regions that are highly conserved among influenza A viruses, and has therefore been one of the target viral proteins for development of a universal influenza vaccine. It has been expected that human or humanized antibodies will be made available for the prophylaxis, pre-emptive and acute treatment of viral infection. However, it is still unclear whether anti-NP human antibody can confer protection against influenza virus infection. In this study, we generated transgenic mice expressing anti-NP human mAbs derived from lymphocytes of a patient infected with H5N1 highly pathogenic avian influenza (HPAI) virus, and experimental infections were conducted to examine antiviral effects of the anti-NP antibodies against H5N1 HPAI viral infections with a high fatality rate in mammals. Transgenic mouse lines expressing the anti-NP human mAbs at more than 1 mg ml-1 showed marked resistance to H5N1 virus infections. In addition, resistance to infection with an H1N1 subtype that shows strong pathogenicity to mice was also confirmed. Although the anti-NP mAbs expressed in the transgenic mice did not neutralize the virus, the mAbs could bind to NP located on the surface of infected cells. These results suggested a possibility that the non-neutralizing anti-NP human mAbs could induce indirect antiviral effects, such as antibody-dependent cellular cytotoxicity or complement-dependent cytotoxicity. Taken together, these results demonstrated that anti-NP human mAbs play an important role in heterosubtypic protection against lethal influenza virus infections in vivo.


Subject(s)
Antibodies, Monoclonal/immunology , Antibodies, Viral/immunology , Immunologic Factors/immunology , Influenza A Virus, H5N1 Subtype/immunology , Orthomyxoviridae Infections/immunology , RNA-Binding Proteins/immunology , Viral Core Proteins/immunology , Animals , Antibodies, Monoclonal/genetics , Antibodies, Viral/genetics , Disease Models, Animal , Disease Resistance , Humans , Immunologic Factors/genetics , Influenza A Virus, H1N1 Subtype/immunology , Mice , Mice, Transgenic , Nucleocapsid Proteins , RNA-Binding Proteins/genetics , Survival Analysis , Viral Core Proteins/genetics
4.
Vet Microbiol ; 166(1-2): 179-83, 2013 Sep 27.
Article in English | MEDLINE | ID: mdl-23849095

ABSTRACT

In Vietnam, numerous surveillance programs are conducted to monitor the prevalence of avian influenza (AI) viruses. Three serological methods-the agar-gel immunodiffusion test, hemagglutination inhibition (HI) test, and enzyme-linked immunosorbent assay-are well established for detection of AI virus antibodies in poultry sera. Several recent reports have validated egg yolk as an alternative source for detection of AI virus antibodies. In this study, we investigated AI virus antibodies in ducks by HI testing using egg yolk. Ten duck eggs were collected every month from 10 randomly selected markets in Hanoi from April 2010 to March 2012. The HI test was performed using low pathogenic avian influenza (LPAI) viruses (H3, H4, H6, H7, H9, and H11 subtypes) and highly pathogenic avian influenza (HPAI) viruses (H5N1 clade 2.3.4 and 2.3.2.1) as antigens. HI testing for H3, H6, and H9 was 29% positive in November 2010, 50% positive in October and November 2010, and 12% positive in June 2011. These results indicated that several epidemics of LPAI viruses had occurred during the study period. In addition, antibodies against H7 were negative. The results of HI testing for H5N1 showed that the reactivity of the dominant HI antibody shifted from H5N1 clade 2.3.4 to clade 2.3.2.1. In conclusion, egg yolk is useful for long term monitoring of AI virus antibodies and the use of egg-based antibody detection may contribute to improvements in animal welfare.


Subject(s)
Antibodies, Viral/immunology , Ducks/immunology , Egg Yolk/immunology , Influenza A virus/immunology , Influenza in Birds/immunology , Poultry Diseases/immunology , Animals , Ducks/virology , Egg Yolk/virology , Hemagglutination Inhibition Tests , Influenza A virus/classification , Influenza A virus/genetics , Influenza in Birds/epidemiology , Influenza in Birds/virology , Poultry Diseases/epidemiology , Poultry Diseases/virology , Sentinel Surveillance/veterinary , Vietnam/epidemiology
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