Biological Properties of H5 Hemagglutinin Expressed by Vaccinia Virus Vector and its Immunological Reactivity with Human Sera

Noisumdaeng, Pirom; Pooruk, Phisanu; Kongchanagul, Alita; Assanasen, Susan; Kitphati, Rungrueng; Auewarakul, Prasert; Puthavathana, Pilaipan

doi:10.1089/vim.2012.0055

Cited by 8 publications

(4 citation statements)

References 48 publications

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“…HA and NA genes, which encode major viral surface glycoproteins, are hypervariable and continuously evolve ( Gutiérrez et al, 2009 ). The biological properties and immune responses against HA and NA glycoproteins of these H5N1 viruses have been previously characterized ( Panaampon et al, 2012 ; Noisumdaeng et al, 2013 ; Noisumdaeng et al, 2014 ; Changsom et al, 2016 ; Noisumdaeng et al, 2021 ). This study comprehensively analyzed the genetic evolution of Thai H5N1 HA and NA nucleotide sequences available in the GenBank database from 2003 to 2010.…”

Section: Discussionmentioning

confidence: 99%

Genetic evolution of hemagglutinin and neuraminidase genes of H5N1 highly pathogenic avian influenza viruses in Thailand

Noisumdaeng

Phadungsombat

Weerated

et al. 2022

PeerJ

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Background Ongoing outbreaks of H5N1 highly pathogenic avian influenza (HPAI) viruses and the emergence of the genetic-related hemagglutinin (HA) gene of reassortant H5Nx viruses currently circulating in wild birds and poultries pose a great global public health concern. In this study, we comprehensively analyzed the genetic evolution of Thai H5N1 HA and neuraminidase (NA) genes between 2003 and 2010. The H5N1 Thailand virus clade 2.3.4 was also genetically compared to the currently circulating clade 2.3.4.4 of H5Nx viruses. Methods Full-length nucleotide sequences of 178 HA and 143 NA genes of H5N1 viruses circulating between 2003 and 2010 were phylogenetically analyzed using maximum likelihood (ML) phylogenetic construction. Bayesian phylogenetic trees were reconstructed using BEAST analysis with a Bayesian Markov chain Monte Carlo (MCMC) approach. The maximum clade credibility (MCC) tree was determined, and the time of the most recent common ancestor (tMRCA) was estimated. The H5N1 HA nucleotide sequences of clade 2.3.4 Thailand viruses were phylogenetically analyzed using ML phylogenetic tree construction and analyzed for nucleotide similarities with various subtypes of reassortant H5Nx HA clade 2.3.4.4. Results ML phylogenetic analysis revealed two distinct HA clades, clade 1 and clade 2.3.4, and two distinct NA groups within the corresponding H5 clade 1 viruses. Bayesian phylogenetic reconstruction for molecular clock suggested that the Thai H5N1 HA and NA emerged in 2001.87 (95% HPD: 2001.34-2002.49) and 2002.38 (95% HPD: 2001.99-2002.82), respectively, suggesting that the virus existed before it was first reported in 2004. The Thai H5N1 HA clade 2.3.4 was grouped into corresponding clades 2.3.4, 2.3.4.1, 2.3.4.2, and 2.3.4.3, and shared nucleotide similarities to reassortant H5Nx clade 2.3.4.4 ranged from 92.4-96.8%. Phylogenetic analysis revealed monophyletic H5Nx clade 2.3.4.4 evolved from H5N1 clade 2.3.4. Conclusion H5N1 viruses existed, and were presumably introduced and circulated in avian species in Thailand, before they were officially reported in 2004. HA and NA genes continuously evolved during circulation between 2004 and 2010. This study provides a better understanding of genetic evolution with respect to molecular epidemiology. Monitoring and surveillance of emerging variants/reassortants should be continued.

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Section: Discussionmentioning

confidence: 99%

Genetic evolution of hemagglutinin and neuraminidase genes of H5N1 highly pathogenic avian influenza viruses in Thailand

Noisumdaeng

Phadungsombat

Weerated

et al. 2022

PeerJ

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“…Sera, plasma and peripheral blood mononuclear cells (PBMCs) were obtained from 37 participants including four H5N1 Thai survivors, 10 H3N2 patients and 23 healthy individuals (Kitphati et al, 2009;Noisumdaeng et al, 2013;Noisumdaeng et al, 2014). Survivor nos.…”

Section: Human Subjects and Blood Specimenmentioning

confidence: 99%

“…Recombinant vaccinia virus carrying H5N1 NP gene inserted in the pSC11 plasmid backbone (rVac-NP virus) or the recombinant virus carrying only the pSC11 backbone (rVac-pSC11 virus) were constructed in our laboratory. The viruses were propagated and titrated in Thymidine kinase negative (TK − ) cells maintained in Dulbecco's modified eagle medium (DMEM) (Gibco) supplemented with 10% FBS plus penicillin and streptomycin (Noisumdaeng et al, 2013;Noisumdaeng et al, 2014). The virus stocks were kept at −80 • C until used.…”

Section: Recombinant Vaccinia Virusesmentioning

confidence: 99%

T cell mediated immunity against influenza H5N1 nucleoprotein, matrix and hemagglutinin derived epitopes in H5N1 survivors and non-H5N1 subjects

Noisumdaeng

Roytrakul

Prasertsopon

et al. 2021

PeerJ

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Background Protection against the influenza virus by a specific antibody is relatively strain specific; meanwhile broader immunity may be conferred by cell-mediated immune response to the conserved epitopes across influenza virus subtypes. A universal broad-spectrum influenza vaccine which confronts not only seasonal influenza virus, but also avian influenza H5N1 virus is promising. Methods This study determined the specific and cross-reactive T cell responses against the highly pathogenic avian influenza A (H5N1) virus in four survivors and 33 non-H5N1 subjects including 10 H3N2 patients and 23 healthy individuals. Ex vivo IFN-γ ELISpot assay using overlapping peptides spanning the entire nucleoprotein (NP), matrix (M) and hemagglutinin (HA) derived from A/Thailand/1(KAN-1)/2004 (H5N1) virus was employed in adjunct with flow cytometry for determining T cell functions. Microneutralization (microNT) assay was performed to determine the status of previous H5N1 virus infection. Results IFN-γ ELISpot assay demonstrated that survivors nos. 1 and 2 had markedly higher T cell responses against H5N1 NP, M and HA epitopes than survivors nos. 3 and 4; and the magnitude of T cell responses against NP were higher than that of M and HA. Durability of the immunoreactivity persisted for as long as four years after disease onset. Upon stimulation by NP in IFN-γ ELISpot assay, 60% of H3N2 patients and 39% of healthy subjects exhibited a cross-reactive T cell response. The higher frequency and magnitude of responses in H3N2 patients may be due to blood collection at the convalescent phase of the patients. In H5N1 survivors, the effector peptide-specific T cells generated from bulk culture PBMCs by in vitro stimulation displayed a polyfunction by simultaneously producing IFN-γ and TNF-α, together with upregulation of CD107a in recognition of the target cells pulsed with peptide or infected with rVac-NP virus as investigated by flow cytometry. Conclusions This study provides an insight into the better understanding on the homosubtypic and heterosubtypic T cell-mediated immune responses in H5N1 survivors and non-H5N1 subjects. NP is an immunodominant target of cross-recognition owing to its high conservancy. Therefore, the development of vaccine targeting the conserved NP may be a novel strategy for influenza vaccine design.

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“…Many laboratories have initiated research on the use of naturally evolved strains for vaccination, especially research on improving the safety of VV and other poxviruses. In particular, many studies have shown that VV is useful for the study of vaccine vectors and exogenous gene expression systems (Garcia-Arriaza et al, 2013 ; Noisumdaeng et al, 2013 ; Adelfinger et al, 2014 ; Remy-Ziller et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Generation of an Attenuated Tiantan Vaccinia Virus Strain by Deletion of Multiple Genes

Zhu

Chen

et al. 2017

Front. Cell. Infect. Microbiol.

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An attenuated vaccinia virus-MVTTEAB-was constructed by deletion of non-essential gene segments related to the immunomodulatory and virulence functions of the vaccinia virus Tiantan strain (VVTT). The shuttle plasmids pTC-EGFP, pTE-EGFP, pTA35-EGFP, pTB-EGFP, and pTA66-EGFP were constructed and combined with the early and late strong promoter pE/L and EGFP as an exogenous selectable marker. Then, through the homologous recombination technology and Cre/loxP system, the following gene fragments were gradually knocked out one by one: TC7L-TK2L, TE3L, TA35R, TB13R, and TA66R. Ultimately, the five-segment-deleted attenuated strain MVTTEAB was obtained. Knockout of these segments and genetic stability of MVTTEAB were confirmed, and it was also shown that knockout of these segments did not affect the replication ability of the virus. Further, a series of in vivo and in vitro experiments demonstrated that the virulence of MVTTEAB was attenuated significantly, but at same time, high immunogenicity was maintained. These results indicate that MVTTEAB has potential for clinical use as a safe viral vector or vaccine with good attenuation and immunogenicity.

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Biological Properties of H5 Hemagglutinin Expressed by Vaccinia Virus Vector and its Immunological Reactivity with Human Sera

Cited by 8 publications

References 48 publications

Genetic evolution of hemagglutinin and neuraminidase genes of H5N1 highly pathogenic avian influenza viruses in Thailand

Genetic evolution of hemagglutinin and neuraminidase genes of H5N1 highly pathogenic avian influenza viruses in Thailand

T cell mediated immunity against influenza H5N1 nucleoprotein, matrix and hemagglutinin derived epitopes in H5N1 survivors and non-H5N1 subjects

Generation of an Attenuated Tiantan Vaccinia Virus Strain by Deletion of Multiple Genes

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