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1.
J Virol ; 92(5)2018 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-29212944

RESUMO

Human herpesvirus 6A (HHV-6A) glycoprotein B (gB) is a glycoprotein consisting of 830 amino acids and is essential for the growth of the virus. Previously, we reported that a neutralizing monoclonal antibody (MAb) called 87-y-13 specifically reacts with HHV-6A gB, and we identified its epitope residue at asparagine (Asn) 347 on gB. In this study, we examined whether the epitope recognized by the neutralizing MAb is essential for HHV-6A infection. We constructed HHV-6A bacterial artificial chromosome (BAC) genomes harboring substitutions at Asn347, namely, HHV-6A BACgB(N347K) and HHV-6A BACgB(N347A). These mutant viruses could be reconstituted and propagated in the same manner as the wild type and their revertants, and MAb 87-y-13 could not inhibit infection by either mutant. In a cell-cell fusion assay, Asn at position 347 on gB was found to be nonessential for cell-cell fusion. In addition, in building an HHV-6A gB homology model, we found that the epitope of the neutralizing MAb is located on domain II of gB and is accessible to solvents. These results indicate that Asn at position 347, the linear epitope of the neutralizing MAb, does not affect HHV-6A infectivity.IMPORTANCE Glycoprotein B (gB) is one of the most conserved glycoproteins among all herpesviruses and is a key factor for virus entry. Therefore, antibodies targeted to gB may neutralize virus entry. Human herpesvirus 6A (HHV-6A) encodes gB, which is translated to a protein of about 830 amino acids (aa). Using a monoclonal antibody (MAb) for HHV-6A gB, which has a neutralizing linear epitope, we analyzed the role of its epitope residue, N347, in HHV-6A infectivity. Interestingly, this gB linear epitope residue, N347, was not essential for HHV-6A growth. By constructing a homology model of HHV-6A gB, we found that N347 was located in the region corresponding to domain II. Therefore, with regard to its neutralizing activity against HHV-6A infection, the epitope on gB might be exposed to solvents, suggesting that it might be a target of the immune system.


Assuntos
Anticorpos Neutralizantes/imunologia , Epitopos/imunologia , Herpesvirus Humano 6/imunologia , Proteínas do Envelope Viral/imunologia , Proteínas Virais de Fusão/imunologia , Sequência de Aminoácidos , Substituição de Aminoácidos/imunologia , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Anticorpos Neutralizantes/metabolismo , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/metabolismo , Fusão Celular , Linhagem Celular , Glicoproteínas/imunologia , Células HEK293 , Herpesviridae/química , Herpesviridae/imunologia , Herpesvirus Humano 6/genética , Herpesvirus Humano 6/crescimento & desenvolvimento , Herpesvirus Humano 6/patogenicidade , Humanos , Proteína Cofatora de Membrana/metabolismo , Mutação , Testes de Neutralização , Domínios Proteicos/imunologia , Proteínas Recombinantes , Análise de Sequência de Proteína , Linfócitos T , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/metabolismo , Proteínas Virais de Fusão/química , Proteínas Virais de Fusão/metabolismo , Internalização do Vírus
2.
OMICS ; 23(11): 573-582, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31651219

RESUMO

Microbial communities residing in the nose play important roles in human health and disease. We report marked differences in nasal microbiota between a rural community and an industrial setting located near a major urban city. Nasal samples were collected from 19 healthy male subjects: 9 samples from persons living in a rural village, and 10 samples from ceramic factory workers in a major industrial Egyptian city. The nasal microbiota in the rural sample had higher and distinct diversity compared with industrial samples from workers exposed to pollution daily. Taxonomic analysis of the sequences revealed five major phyla; among these phyla were Actinobacteria, Proteobacteria, Bacteroidetes, and Fusobacteria, revealing significant abundance variation by geographical location. For example, the rural group had a significant increase in representation of Actinobacteria and Bacteroidetes (p = 0.004, p = 0.01, respectively) compared with the industrial group. However, the industrial group showed a significant increase in relative abundance of phylum Proteobacteria (p = 0.02). The most predominant genera for the rural group were Corynebacterium, Staphylococcus, Alloiococcus, and Peptoniphilus. By contrast, the industrial group was dominated by Staphylococcus, Sphingomonas, and Moraxella. Environmental pollution might alter the nasal microbiome leading to an attendant disturbance in the microbiome community structure. The clinical and public health implications of these nasal microbiome variations by rural and industrialized geography warrant further research. This study contributes to our knowledge of the bacterial composition of nasal microbiome in rural and industrialized geographies, and informs public health, respiratory medicine, and occupational health scholarship.


Assuntos
Metagenômica , Microbiota , Mucosa Nasal/microbiologia , Exposição Ocupacional , População Rural , Bactérias/classificação , Bactérias/genética , Biodiversidade , Código de Barras de DNA Taxonômico , Egito , Humanos , Indústrias , Metagenômica/métodos , Vigilância em Saúde Pública , RNA Ribossômico 16S
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