scholarly journals Enhanced isolation of influenza viruses in qualified cells improves the probability of well-matched vaccines

npj Vaccines ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Heidi Peck ◽  
Karen L. Laurie ◽  
Steve Rockman ◽  
Vivian Leung ◽  
Hilda Lau ◽  
...  
Keyword(s):  
Seasonal Influenza ◽  
Virus Isolation ◽  
Growth Characteristics ◽  
Influenza Viruses ◽  
Influenza Vaccines ◽  
Isolation Method ◽  
Culture Production ◽  
Ha Protein ◽  
Chicken Eggs ◽  
Embryonated Chicken

AbstractInfluenza vaccines are utilised to combat seasonal and pandemic influenza. The key to influenza vaccination currently is the availability of candidate vaccine viruses (CVVs). Ideally, CVVs reflect the antigenic characteristics of the circulating virus, which may vary depending upon the isolation method. For traditional inactivated egg-based vaccines, CVVs are isolated in embryonated chicken eggs, while for cell-culture production, CVV’s are isolated in either embryonated eggs or qualified cell lines. We compared isolation rates, growth characteristics, genetic stability and antigenicity of cell and egg CVV’s derived from the same influenza-positive human clinical respiratory samples collected from 2008–2020. Influenza virus isolation rates in MDCK33016PF cells were twice that of eggs and mutations in the HA protein were common in egg CVVs but rare in cell CVVs. These results indicate that fully cell-based influenza vaccines will improve the choice, match and potentially the effectiveness, of seasonal influenza vaccines compared to egg-based vaccines.

scholarly journals Molecular Characterization of Seasonal Influenza A and B from Hospitalized Patients in Thailand in 2018–2019

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 977
Author(s):  
Kobporn Boonnak ◽  
Chayasin Mansanguan ◽  
Dennis Schuerch ◽  
Usa Boonyuen ◽  
Hatairat Lerdsamran ◽  
...  
Keyword(s):  
Amino Acid ◽  
Influenza A ◽  
Seasonal Influenza ◽  
Influenza Viruses ◽  
Surface Proteins ◽  
Antigenic Drift ◽  
Influenza B ◽  
Receptor Binding Site ◽  
Antigenic Sites ◽  
Ha Protein

Influenza viruses continue to be a major public health threat due to the possible emergence of more virulent influenza virus strains resulting from dynamic changes in virus adaptability, consequent of functional mutations and antigenic drift in surface proteins, especially hemagglutinin (HA) and neuraminidase (NA). In this study, we describe the genetic and evolutionary characteristics of H1N1, H3N2, and influenza B strains detected in severe cases of seasonal influenza in Thailand from 2018 to 2019. We genetically characterized seven A/H1N1 isolates, seven A/H3N2 isolates, and six influenza B isolates. Five of the seven A/H1N1 viruses were found to belong to clade 6B.1 and were antigenically similar to A/Switzerland/3330/2017 (H1N1), whereas two isolates belonged to clade 6B.1A1 and clustered with A/Brisbane/02/2018 (H1N1). Interestingly, we observed additional mutations at antigenic sites (S91R, S181T, T202I) as well as a unique mutation at a receptor binding site (S200P). Three-dimensional (3D) protein structure analysis of hemagglutinin protein reveals that this unique mutation may lead to the altered binding of the HA protein to a sialic acid receptor. A/H3N2 isolates were found to belong to clade 3C.2a2 and 3C.2a1b, clustering with A/Switzerland/8060/2017 (H3N2) and A/South Australia/34/2019 (H3N2), respectively. Amino acid sequence analysis revealed 10 mutations at antigenic sites including T144A/I, T151K, Q213R, S214P, T176K, D69N, Q277R, N137K, N187K, and E78K/G. All influenza B isolates in this study belong to the Victoria lineage. Five out of six isolates belong to clade 1A3-DEL, which relate closely to B/Washington/02/2009, with one isolate lacking the three amino acid deletion on the HA segment at position K162, N163, and D164. In comparison to the B/Colorado/06/2017, which is the representative of influenza B Victoria lineage vaccine strain, these substitutions include G129D, G133R, K136E, and V180R for HA protein. Importantly, the susceptibility to oseltamivir of influenza B isolates, but not A/H1N1 and A/H3N2 isolates, were reduced as assessed by the phenotypic assay. This study demonstrates the importance of monitoring genetic variation in influenza viruses regarding how acquired mutations could be associated with an improved adaptability for efficient transmission.

scholarly journals Development and Evaluation of Vero Cell-Derived Master Donor Viruses for Influenza Pandemic Preparedness

Vaccines ◽  
2020 ◽  
Vol 8 (4) ◽  
pp. 626
Author(s):  
Po-Ling Chen ◽  
Tsai-Teng Tzeng ◽  
Alan Yung-Chih Hu ◽  
Lily Hui-Ching Wang ◽  
Min-Shi Lee
Keyword(s):  
Vero Cell ◽  
Seasonal Influenza ◽  
Influenza Pandemic ◽  
High Growth ◽  
Vero Cells ◽  
Influenza Viruses ◽  
Influenza Vaccines ◽  
Pandemic Preparedness ◽  
Influenza Pandemics ◽  
Production Platform

The embryonated egg-based platform currently produces the majority of seasonal influenza vaccines by employing a well-developed master donor virus (MDV, A/PR/8/34 (PR8)) to generate high-growth reassortants (HGRs) for A/H1N1 and A/H3N2 subtypes. Although the egg-based platform can supply enough seasonal influenza vaccines, it cannot meet surging demands during influenza pandemics. Therefore, multi-purpose platforms are desirable for pandemic preparedness. The Vero cell-based production platform is widely used for human vaccines and could be a potential multi-purpose platform for pandemic influenza vaccines. However, many wild-type and egg-derived influenza viruses cannot grow efficiently in Vero cells. Therefore, it is critical to develop Vero cell-derived high-growth MDVs for pandemic preparedness. In this study, we evaluated two in-house MDVs (Vero-15 and VB5) and two external MDVs (PR8 and PR8-HY) to generate Vero cell-derived HGRs for five avian influenza viruses (AIVs) with pandemic potentials (H5N1 clade 2.3.4, H5N1 clade 2.3.2.1, American-lineage H5N2, H7N9 first wave and H7N9 fifth wave). Overall, no single MDV could generate HGRs for all five AIVs, but this goal could be achieved by employing two in-house MDVs (vB5 and Vero-15). In immunization studies, mice received two doses of Vero cell-derived inactivated H5N1 and H7N9 whole virus antigens adjuvanted with alum and developed robust antibody responses.

scholarly journals Improvement of Influenza A/Fujian/411/02 (H3N2) Virus Growth in Embryonated Chicken Eggs by Balancing the Hemagglutinin and Neuraminidase Activities, Using Reverse Genetics

2005 ◽  
Vol 79 (11) ◽  
pp. 6763-6771 ◽  
Author(s):  
Bin Lu ◽  
Helen Zhou ◽  
Dan Ye ◽  
George Kemble ◽  
Hong Jin
Keyword(s):  
Amino Acids ◽  
Virus Replication ◽  
Receptor Binding ◽  
Influenza A ◽  
Reverse Genetics ◽  
Influenza Vaccines ◽  
H3n2 Virus ◽  
Virus Growth ◽  
Chicken Eggs ◽  
Embryonated Chicken

ABSTRACT The H3N2 influenza A/Fujian/411/02-like virus strains that circulated during the 2003-2004 influenza season caused influenza epidemics. Most of the A/Fujian/411/02 virus lineages did not replicate well in embryonated chicken eggs and had to be isolated originally by cell culture. The molecular basis for the poor replication of A/Fujian/411/02 virus was examined in this study by the reverse genetics technology. Two antigenically related strains that replicated well in embryonated chicken eggs, A/Sendai-H/F4962/02 and A/Wyoming/03/03, were compared with the prototype A/Fujian/411/02 virus. A/Sendai differed from A/Fujian by three amino acids in the neuraminidase (NA), whereas A/Wyoming differed from A/Fujian by five amino acids in the hemagglutinin (HA). The HA and NA segments of these three viruses were reassorted with cold-adapted A/Ann Arbor/6/60, the master donor virus for the live attenuated type A influenza vaccines (FluMist). The HA and NA residues differed between these three H3N2 viruses evaluated for their impact on virus replication in MDCK cells and in embryonated chicken eggs. It was determined that replication of A/Fujian/411/02 in eggs could be improved by either changing minimum of two HA residues (G186V and V226I) to increase the HA receptor-binding ability or by changing a minimum of two NA residues (E119Q and Q136K) to lower the NA enzymatic activity. Alternatively, recombinant A/Fujian/411/02 virus could be adapted to grow in eggs by two amino acid substitutions in the HA molecule (H183L and V226A), which also resulted in the increased HA receptor-binding activity. Thus, the balance between the HA and NA activities is critical for influenza virus replication in a different host system. The HA or NA changes that increased A/Fujian/411/02 virus replication in embryonated chicken eggs were found to have no significant impact on antigenicity of these recombinant viruses. This study demonstrated that the reverse genetics technology could be used to improve the manufacture of the influenza vaccines.

scholarly journals An Overview of Influenza Viruses and Vaccines

Vaccines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1032
Author(s):  
Rina Fajri Nuwarda ◽  
Abdulsalam Abdullah Alharbi ◽  
Veysel Kayser
Keyword(s):  
Seasonal Influenza ◽  
Vaccine Development ◽  
Influenza Infection ◽  
Influenza Viruses ◽  
Influenza Vaccines ◽  
Efficacy And Safety ◽  
Testing Methods ◽  
Health Concerns ◽  
Global Pandemic ◽  
And Control

Influenza remains one of the major public health concerns because it causes annual epidemics and can potentially instigate a global pandemic. Numerous countermeasures, including vaccines and antiviral treatments, are in use against seasonal influenza infection; however, their effectiveness has always been discussed due to the ongoing resistance to antivirals and relatively low and unpredictable efficiency of influenza vaccines compared to other vaccines. The growing interest in vaccines as a promising approach to prevent and control influenza may provide alternative vaccine development options with potentially increased efficiency. In addition to currently available inactivated, live-attenuated, and recombinant influenza vaccines on the market, novel platforms such as virus-like particles (VLPs) and nanoparticles, and new vaccine formulations are presently being explored. These platforms provide the opportunity to design influenza vaccines with improved properties to maximize quality, efficacy, and safety. The influenza vaccine manufacturing process is also moving forward with advancements relating to egg- and cell-based production, purification processes, and studies into the physicochemical attributes and vaccine degradation pathways. These will contribute to the design of more stable, optimized vaccine formulations guided by contemporary analytical testing methods and via the implementation of the latest advances in the field.

An Epidemiological Study of Sheep and Goat pox Outbreaks in the Sudan

Food Biology ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Ahmed Zein Elabdeen Mahmoud ◽  
Abdelmalik Ibrahim Khalafalla ◽  
Muaz Magzob Abdellatif
Keyword(s):  
Virus Isolation ◽  
Chorioallantoic Membrane ◽  
Vero Cells ◽  
Fibroblast Cells ◽  
Chick Embryo Fibroblast ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Species Specific ◽  
Chicken Eggs ◽  
Embryonated Chicken

Sheep and goat pox Outbreaks occurred in different geographic areas of Sudan and most strikingly, were highly species specific. Two outbreaks in Gedarif State in June. 2013 affected no goats and outbreak in Khartoum state in March. 2015 affected no sheep despite communal herding; affected goats were vaccinated with 0240 strain. Clinically, the disease was characterized by fever, depression and eruption of generalized pox lesions. Mortality rate ranged between 5.2 and 6.7% with a mean of 6.1%. Isolation of viruses succeed on Lamb testes cell culture at passage four, the diseases were diagnosed using virus neutralisation test and polymerase chain reaction. Sheeppox and goatpox isolates grew well in lamb testes and Vero cells. In MDBK however, both viruses induced slight CPE that reached 60% in 9 days. On the other hand, both isolates induced no CPE in chick embryo fibroblast cells. Virus isolation attempts failed on chorioallantoic membrane of embryonated chicken eggs.

scholarly journals Prolonged Protection against Intranasal Challenge with Influenza Virus following Systemic Immunization or Combinations of Mucosal and Systemic Immunizations with a Heat-Labile Toxin Mutant

2009 ◽  
Vol 16 (4) ◽  
pp. 471-478 ◽  
Author(s):  
Fengmin Zhou ◽  
Amanda Goodsell ◽  
Yasushi Uematsu ◽  
Michael Vajdy
Keyword(s):  
Influenza Virus ◽  
Seasonal Influenza ◽  
Lethal Dose ◽  
Influenza Viruses ◽  
Influenza Vaccines ◽  
Virus Infections ◽  
Vaccination Strategies ◽  
Heat Labile ◽  
The World ◽  
Considerable Morbidity

ABSTRACTSeasonal influenza virus infections cause considerable morbidity and mortality in the world, and there is a serious threat of a pandemic influenza with the potential to cause millions of deaths. Therefore, practical influenza vaccines and vaccination strategies that can confer protection against intranasal infection with influenza viruses are needed. In this study, we demonstrate that using LTK63, a nontoxic mutant of the heat-labile toxin fromEscherichia coli, as an adjuvant for both mucosal and systemic immunizations, systemic (intramuscular) immunization or combinations of mucosal (intranasal) and intramuscular immunizations protected mice against intranasal challenge with a lethal dose of live influenza virus at 3.5 months after the second immunization.

scholarly journals Serum Samples From Middle-aged Adults Vaccinated Annually with Seasonal Influenza Vaccines Cross-neutralize Some Potential Pandemic Influenza Viruses

2015 ◽  
Vol 213 (3) ◽  
pp. 403-406 ◽  
Author(s):  
Wei Wang ◽  
Esmeralda Alvarado-Facundo ◽  
Qiong Chen ◽  
Christine M. Anderson ◽  
Dorothy Scott ◽  
...  
Keyword(s):  
Pandemic Influenza ◽  
Seasonal Influenza ◽  
Influenza Viruses ◽  
Influenza Vaccines ◽  
Middle Aged ◽  
Serum Samples ◽  
Middle Aged Adults
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