Live influenza vaccine: screening of attenuated virus strains by a 50% ciliary activity inhibition test in organ cultures of ferret trachea

A. Boudreault

doi:10.1139/m79-045

Live influenza vaccine: screening of attenuated virus strains by a 50% ciliary activity inhibition test in organ cultures of ferret trachea

Canadian Journal of Microbiology ◽

10.1139/m79-045 ◽

1979 ◽

Vol 25 (3) ◽

pp. 279-284 ◽

Cited By ~ 3

Author(s):

A. Boudreault

Keyword(s):

Influenza Vaccine ◽

Influenza Vaccines ◽

Ciliary Activity ◽

Organ Cultures ◽

Activity Inhibition ◽

Attenuated Virus ◽

Time Required ◽

Virus Strains ◽

Better Than

This study of three live attenuated inhibitor-resistant influenza vaccines showed that these preparations are usually antigenic and that they caused no significant reactions when characterized by an index of attenuation equal to or slightly better than 1.0 arbitrarily attributed to the 'reference' attenuated A/Hong Kong/68 strain of Beare and Bynoe. This index, measured in vitro on ferret tracheal rings, is expressed as the ratio of the time required for ciliary activity inhibition of 50% of the rings by the tested candidate vaccine strain and the 'reference' attenuated strain. Induction of heterologous antibodies was also observed. Oral administration of underattenuated preparations did not cause the severe reactions which were observed when the same vaccine was administered intranasally.

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Inhibition of ciliary activity in organ cultures of ferret trachea in reference to genetic and biological characters of influenza virus strains

Canadian Journal of Microbiology ◽

10.1139/m82-122 ◽

1982 ◽

Vol 28 (7) ◽

pp. 809-814 ◽

Cited By ~ 3

Author(s):

P. Diaz-Rodriguez ◽

A. Boudreault

Keyword(s):

Influenza Virus ◽

Influenza Viruses ◽

Influenza Vaccines ◽

Ciliary Activity ◽

Organ Cultures ◽

Preliminary Screening ◽

Cold Adapted ◽

Activity Inhibition ◽

Biological Characters ◽

Virus Strains

As reported previously, attenuated stable inhibitor-resistant influenza viruses can be screened by a 50% ciliary activity inhibition test in ferret tracheal organ cultures. This test was further applied to 5 attenuated cold-adapted influenza strains and to 11 strains with known a percentage of RNA–RNA hybridization with the parental A/PR/8/34 (H0N1) virus strain. Again, with one exception, attenuated strains could be clearly differentiated from virulent ones. It was concluded that virulence of influenza strains for man can be detected using this test regardless of the techniques used to prepare attenuated variants. A preliminary screening of attenuated candidates for live influenza vaccines can be achieved with confidence on ferret tracheal organ cultures.

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Hepatitis a virus: Growth characteristics of in vivo and in vitro propagated wild and attenuated virus strains

Journal of Medical Virology ◽

10.1002/jmv.1890140410 ◽

1984 ◽

Vol 14 (4) ◽

pp. 373-386 ◽

Cited By ~ 17

Author(s):

Daniel W. Bradley ◽

Charles A. Schable ◽

Karen A. McCaustland ◽

E. H. Cook ◽

Bert L. Murphy ◽

...

Keyword(s):

Hepatitis A ◽

Hepatitis A Virus ◽

Growth Characteristics ◽

Virus Growth ◽

Attenuated Virus ◽

Virus Strains

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Tissue Culture of Human Adult Adenoids and of Middle Ear Mucosa

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348947608500303 ◽

1976 ◽

Vol 85 (3) ◽

pp. 327-333 ◽

Cited By ~ 8

Author(s):

Ilana Drucker ◽

Ziva Weisman ◽

Jacob Sadé

Keyword(s):

Middle Ear ◽

Ciliary Activity ◽

Organ Cultures ◽

Middle Ear Mucosa ◽

Monolayer Growth ◽

Stratified Squamous Epithelium ◽

Inducing Factors ◽

Developmental Characteristics ◽

Mucociliary Epithelia

The increased number of mucus producing cells as well as the presence of stratified squamous epithelium in pathological and experimental middle ear conditions, point towards the possibility of metaplastic changes of the middle ear mucosa, similar to the metaplastic capabilities of respiratory mucosae in general, as observed clinically or provoked experimentally. The purpose of this study was to develop a model of postembryonic human respiratory mucosae, in vitro, for the study of triggering or inducing factors involved in its normal and metaplastic differentiation. Explants from adenoids and middle ear mucosa were cultured, both as organ cultures and monolayers, for periods of up to two weeks, and their developmental characteristics were studied and described. Over 50% of the explants showed mitosis, epithelial and monolayer growth, ciliary activity and differentiation into ciliated and into mucus-producing cells. Adenoid explants were grown in air without and with added 5% CO2. Under the latter conditions, the proportion of explants and monolayers showing ciliary activity was 50% greater. It is concluded that this model might be suitable for further studies of the factors which control cyto-differentiation in mucociliary epithelia. Maintaining its growth for a longer period would, however, be desirable.

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The MVA-NP+M Influenza Vaccine did work. It may help defeat COVID.

10.31219/osf.io/sb4zp ◽

2020 ◽

Author(s):

John Stuart Weatherston Gillis

Keyword(s):

Influenza Vaccine ◽

Original Study ◽

Influenza Vaccines ◽

Second Wave ◽

Better Than

Many scientists are developing COVID-19 vaccines. However, we need effective influenza vaccines as well. Otherwise, a second wave of COVID will be a double whammy.Here I show that the results of Davenport et al [1] are significantly better than reported in the original study.

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A Live Attenuated Influenza Vaccine Elicits Enhanced Heterologous Protection When the Internal Genes of the Vaccine Are Matched to Those of the Challenge Virus

Journal of Virology ◽

10.1128/jvi.01065-19 ◽

2019 ◽

Vol 94 (4) ◽

Cited By ~ 3

Author(s):

Andrew Smith ◽

Laura Rodriguez ◽

Maya El Ghouayel ◽

Aitor Nogales ◽

Jeffrey M. Chamberlain ◽

...

Keyword(s):

Influenza Vaccine ◽

Influenza A ◽

H1n1 Virus ◽

Influenza Infection ◽

Influenza Viruses ◽

Influenza Vaccines ◽

Temperature Sensitive ◽

Live Attenuated Influenza Vaccine ◽

Cell Immunity

ABSTRACT Influenza A virus (IAV) causes significant morbidity and mortality, despite the availability of viral vaccines. The efficacy of live attenuated influenza vaccines (LAIVs) has been especially poor in recent years. One potential reason is that the master donor virus (MDV), on which all LAIVs are based, contains either the internal genes of the 1960 A/Ann Arbor/6/60 or the 1957 A/Leningrad/17/57 H2N2 viruses (i.e., they diverge considerably from currently circulating strains). We previously showed that introduction of the temperature-sensitive (ts) residue signature of the AA/60 MDV into a 2009 pandemic A/California/04/09 H1N1 virus (Cal/09) results in only 10-fold in vivo attenuation in mice. We have previously shown that the ts residue signature of the Russian A/Leningrad/17/57 H2N2 LAIV (Len LAIV) more robustly attenuates the prototypical A/Puerto Rico/8/1934 (PR8) H1N1 virus. In this work, we therefore introduced the ts signature from Len LAIV into Cal/09. This new Cal/09 LAIV is ts in vitro, highly attenuated (att) in mice, and protects from a lethal homologous challenge. In addition, when our Cal/09 LAIV with PR8 hemagglutinin and neuraminidase was used to vaccinate mice, it provided enhanced protection against a wild-type Cal/09 challenge relative to a PR8 LAIV with the same attenuating mutations. These findings suggest it may be possible to improve the efficacy of LAIVs by better matching the sequence of the MDV to currently circulating strains. IMPORTANCE Seasonal influenza infection remains a major cause of disease and death, underscoring the need for improved vaccines. Among current influenza vaccines, the live attenuated influenza vaccine (LAIV) is unique in its ability to elicit T-cell immunity to the conserved internal proteins of the virus. Despite this, LAIV has shown limited efficacy in recent years. One possible reason is that the conserved, internal genes of all current LAIVs derive from virus strains that were isolated between 1957 and 1960 and that, as a result, do not resemble currently circulating influenza viruses. We have therefore developed and tested a new LAIV, based on a currently circulating pandemic strain of influenza. Our results show that this new LAIV elicits improved protective immunity compared to a more conventional LAIV.

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Development of a Pan-H1 Influenza Vaccine

Journal of Virology ◽

10.1128/jvi.01349-18 ◽

2018 ◽

Vol 92 (22) ◽

Cited By ~ 15

Author(s):

Nicole Darricarrère ◽

Svetlana Pougatcheva ◽

Xiaochu Duan ◽

Rebecca S. Rudicell ◽

Te-Hui Chou ◽

...

Keyword(s):

Influenza Virus ◽

Immune Responses ◽

Influenza Vaccine ◽

Seasonal Influenza ◽

Influenza Viruses ◽

Influenza Vaccines ◽

The Past ◽

Individual Strain ◽

Humoral Responses ◽

Virus Strains

ABSTRACT The efficacy of current seasonal influenza vaccines varies greatly, depending on the match to circulating viruses. Although most vaccines elicit strain-specific responses, some present cross-reactive epitopes that elicit antibodies against diverse viruses and remain unchanged and effective for several years. To determine whether combinations of specific H1 hemagglutinin (HA) antigens stimulate immune responses that protect against diverse H1 influenza viruses, we evaluated the antibody responses elicited by HA-ferritin nanoparticles derived from six evolutionarily divergent H1 sequences and two computationally optimized broadly reactive antigen (COBRA) HA antigens. Humoral responses were assessed against a panel of 16 representative influenza virus strains from the past 80 years. HAs from the strains A/NewCaledonia/20/1999 (NC99), A/California/04/2009 (CA09), A/HongKong/117/1977 (HK77), COBRA X6, or P1 elicited neutralization against diverse strains, and a combination of three wild-type HA or two COBRA HA nanoparticles conferred significant additional breadth beyond that observed with any individual strain. Therefore, combinations of H1 HAs may constitute a pan-H1 influenza vaccine. IMPORTANCE Seasonal influenza vaccines elicit strain-specific immune responses designed to protect against circulating viruses. Because these vaccines often show limited efficacy, the search for a broadly protective seasonal vaccine remains a priority. Among different influenza virus subtypes, H1N1 has long been circulating in humans and has caused pandemic outbreaks. In order to assess the potential of a multivalent HA combination vaccine to improve the breadth of protection against divergent H1N1 viruses, HA-ferritin nanoparticles were made and evaluated in mice against a panel of historical and contemporary influenza virus strains. Trivalent combinations of H1 nanoparticles improved the breadth of immunity against divergent H1 influenza viruses.

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Ciliostatic, hemagglutinating, and proteolytic activities in a cell extract of Mycoplasma pneumoniae

Infection and Immunity ◽

10.1128/iai.29.3.1111-1116.1980 ◽

1980 ◽

Vol 29 (3) ◽

pp. 1111-1116

Author(s):

D K Chandler ◽

M F Barile

Keyword(s):

Sodium Dodecyl Sulfate ◽

Mycoplasma Pneumoniae ◽

Sodium Dodecyl ◽

Cell Extract ◽

Ciliary Activity ◽

Organ Cultures ◽

Freeze Thaw ◽

A Cell ◽

Proteolytic Activities

An extract of Mycoplasma pneumoniae, prepared from glass-grown organisms by extraction with 2 M NaCl, followed by freeze-thaw, ultracentrifugation, dialysis, and lyophilization, yielded approximately 20% of the total mycoplasmal protein. The extract contained at least 20 protein bands on sodium dodecyl sulfate-polyacrylamide gels and 2 to 5% carbohydrate and inhibited 70 to 100% of the ciliary activity of hamster tracheal organ cultures (ciliostasis). The extent of ciliostasis was dependent on the concentration of the extract. The extract also produced hemagglutination of human O-positive erythrocytes and showed proteolytic activity with a synthetic tetrapeptide substrate, S-2222. These in vitro tissue-damaging activities may be associated with the virulence of the mycoplasmas and with the pathogenesis of M. pneumoniae disease.

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Elicitation of broadly protective immunity to influenza by multivalent hemagglutinin nanoparticle vaccines

10.1101/2020.05.30.125179 ◽

2020 ◽

Cited By ~ 5

Author(s):

Seyhan Boyoglu-Barnum ◽

Daniel Ellis ◽

Rebecca A. Gillespie ◽

Geoffrey B. Hutchinson ◽

Young-Jun Park ◽

...

Keyword(s):

Influenza Vaccine ◽

Neutralizing Antibodies ◽

Protective Immunity ◽

Neutralizing Antibody ◽

Influenza Viruses ◽

Antibody Responses ◽

Influenza Vaccines ◽

Next Generation ◽

Protective Antibody

AbstractInfluenza vaccines that confer broad and durable protection against diverse virus strains would have a major impact on global health. However, next-generation vaccine design efforts have been complicated by challenges including the genetic plasticity of the virus and the immunodominance of certain epitopes in its glycoprotein antigens. Here we show that computationally designed, two-component nanoparticle immunogens induce potently neutralizing and broadly protective antibody responses against a wide variety of influenza viruses. The nanoparticle immunogens display 20 hemagglutinin (HA) trimers in a highly immunogenic array, and their assembly in vitro enables precisely controlled co-display of multiple distinct HAs in defined ratios. Nanoparticle immunogens displaying the four HAs of licensed quadrivalent influenza vaccines (QIV) elicited hemagglutination inhibition and neutralizing antibody responses to vaccine-matched strains that were equivalent or superior to commercial QIV in mice, ferrets, and nonhuman primates. The nanoparticle immunogens—but not QIV—simultaneously induced broadly protective antibody responses to heterologous viruses, including H5N1 and H7N9, by targeting the subdominant yet conserved HA stem. Unlike previously reported influenza vaccine candidates, our nanoparticle immunogens can alter the intrinsic immunodominance hierarchy of HA to induce both potent receptor-blocking and broadly cross-reactive stem-directed antibody responses and are attractive candidates for a next-generation influenza vaccine that could replace current seasonal vaccines.One Sentence SummaryNanoparticle immunogens displaying four seasonal influenza hemagglutinins elicit neutralizing antibodies directed at both the immunodominant head and the conserved stem and confer broad protective immunity.

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