Name: influenza a virus studies in a mouse model of infection
Uploaded: 2017-09-07T13:00:00.000+00:00
Duration: 10 min 44 s
Description: Watch this Scientific Journal Video about Influenza A Virus Studies in a Mouse Model of Infection at JoVE.com

Investigación sobre el virus de la gripe en laboratorio LM-S es financiada parcialmente por la Nueva York Influenza centro de excelencia (NYICE), miembro de la NIAID centros de excelencia para la investigación de la Influenza y vigilancia (CEIRS). Agradecemos a Wendy Bates por su apoyo en las correcciones del manuscrito.

<ol>
	<li>Girard, M. P., Cherian, T., Pervikov, Y., Kieny, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+vaccine+research+and+development:+human+acute+respiratory+infections.">A review of vaccine research and development: human acute respiratory infections.</a> Vaccine. 23 (50), 5708-5724 (2005).</li><li>Arnold, S., Monto, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+and+Virology+of+Influenza+Illness.">Epidemiology and Virology of Influenza Illness.</a> Am J Manag Care. 6, 255-264 (2000).</li><li>Palese, P., Shaw, M. L., Knipe, D. M., Howley, P. M., Griffin, D. E., Lamb, R. A., Martin, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthomyxoviridae:+The+Viruses+and+Their+Replication.">Orthomyxoviridae: The Viruses and Their Replication.</a> Fields Virology. , (2007).</li><li>Li, K. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genesis+of+a+highly+pathogenic+and+potentially+pandemic+H5N1+influenza+virus+in+eastern+Asia.">Genesis of a highly pathogenic and potentially pandemic H5N1 influenza virus in eastern Asia.</a> Nature. 430 (6996), 209-213 (2004).</li><li>Nogales, A., Martinez-Sobrido, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reverse+Genetics+Approaches+for+the+Development+of+Influenza+Vaccines.">Reverse Genetics Approaches for the Development of Influenza Vaccines.</a> Int J Mol Sci. 18 (20), (2017).</li><li>Kunisaki, K. M., Janoff, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influenza+in+immunosuppressed+populations:+a+review+of+infection+frequency,+morbidity,+mortality,+and+vaccine+responses.">Influenza in immunosuppressed populations: a review of infection frequency, morbidity, mortality, and vaccine responses.</a> Lancet Infect Dis. 9 (8), 493-504 (2009).</li><li>Belshe, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+attenuated+versus+inactivated+influenza+vaccine+in+infants+and+young+children.">Live attenuated versus inactivated influenza vaccine in infants and young children.</a> N Engl J Med. 356 (7), 685-696 (2007).</li><li>Cox, A., Baker, S. F., Nogales, A., Martinez-Sobrido, L., Dewhurst, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+mouse-adapted+live+attenuated+influenza+virus+that+permits+in+vivo+analysis+of+enhancements+to+the+safety+of+live+attenuated+influenza+virus+vaccine.">Development of a mouse-adapted live attenuated influenza virus that permits in vivo analysis of enhancements to the safety of live attenuated influenza virus vaccine.</a> J Virol. 89 (6), 3421-3426 (2015).</li><li>Nogales, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influenza+A+Virus+Attenuation+by+Codon+Deoptimization+of+the+NS+Gene+for+Vaccine+Development.">Influenza A Virus Attenuation by Codon Deoptimization of the NS Gene for Vaccine Development.</a> J Virol. 88 (18), 10525-10540 (2014).</li><li>Nogales, A., DeDiego, M. L., Topham, D. J., Martinez-Sobrido, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rearrangement+of+Influenza+Virus+Spliced+Segments+for+the+Development+of+Live-Attenuated+Vaccines.">Rearrangement of Influenza Virus Spliced Segments for the Development of Live-Attenuated Vaccines.</a> J Virol. 90 (14), 6291-6302 (2016).</li><li>Nogales, A., Huang, K., Chauch&#233;, C., DeDiego, M. L., Murcia, P. R., Parrish, C. R., Mart&#237;nez-Sobrido, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Canine+influenza+viruses+with+modified+NS1+proteins+for+the+development+of+live-attenuated+vaccines.">Canine influenza viruses with modified NS1 proteins for the development of live-attenuated vaccines.</a> Virology. 500 (2017), 1-10 (2016).</li><li>Garcia-Sastre, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influenza+A+virus+lacking+the+NS1+gene+replicates+in+interferon-deficient+systems.">Influenza A virus lacking the NS1 gene replicates in interferon-deficient systems.</a> Virology. 252 (2), 324-330 (1998).</li><li>Lowen, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blocking+interhost+transmission+of+influenza+virus+by+vaccination+in+the+guinea+pig+model.">Blocking interhost transmission of influenza virus by vaccination in the guinea pig model.</a> J Virol. 83 (7), 2803-2818 (2009).</li><li>Mubareka, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmission+of+influenza+virus+via+aerosols+and+fomites+in+the+guinea+pig+model.">Transmission of influenza virus via aerosols and fomites in the guinea pig model.</a> J Infect Dis. 199 (6), 858-865 (2009).</li><li>Nogales, A., Baker, S. F., Martinez-Sobrido, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Replication-competent+influenza+A+viruses+expressing+a+red+fluorescent+protein.">Replication-competent influenza A viruses expressing a red fluorescent protein.</a> Virology. 476C, 206-216 (2014).</li><li>Margine, I., Krammer, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+for+Influenza+Viruses:+Implications+for+Universal+Vaccine+Development.">Animal Models for Influenza Viruses: Implications for Universal Vaccine Development.</a> Pathogens. 3 (4), 845-874 (2014).</li><li>Bouvier, N., Lowen, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+for+Influenza+Virus+Pathogenesis+and+Transmission.">Animal Models for Influenza Virus Pathogenesis and Transmission.</a> Viruses. 2 (8), 1530-1563 (2010).</li><li>Pica, N., Iyer, A., Ramos, I., Bouvier, N. M., Fernandez-Sesma, A., Garc&#237;a-Sastre, A., Lowen, A. C., Palese, P., Steel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+DBA.2+mouse+is+susceptible+to+disease+following+infection+with+a+broad,+but+limited,+range+of+influenza+A+and+B+viruses.">The DBA.2 mouse is susceptible to disease following infection with a broad, but limited, range of influenza A and B viruses.</a> J Virol. 85 (23), 12825-12829 (2011).</li><li>Watanabe, H., Numata, K., Ito, T., Takagi, K., Matsukawa, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Innate+immune+response+in+Th1-+and+Th2-dominant+mouse+strains.">Innate immune response in Th1- and Th2-dominant mouse strains.</a> Shock. 22 (5), 460-466 (2004).</li><li>Srivastava, B., Blazejewska, P., Hessmann, M., Bruder, D., Geffers, R., Mauel, S., Gruber, A. D., Schughart, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Host+genetic+background+strongly+influences+the+response+to+influenza+a+virus+infections.">Host genetic background strongly influences the response to influenza a virus infections.</a> PLoS One. 4 (3), e4857 (2009).</li><li>Lowen, A. C., Bouvier, N. M., Steel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmission+in+the+Guinea+Pig+Model.">Transmission in the Guinea Pig Model.</a> Curr Top Microbiol Immunol. 385, 157-183 (2014).</li><li>Schickli, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasmid-only+rescue+of+influenza+A+virus+vaccine+candidates.">Plasmid-only rescue of influenza A virus vaccine candidates.</a> Philos Trans R Soc Lond B Biol Sci. 356 (1416), 1965-1973 (2001).</li><li>Martinez-Sobrido, L., Garcia-Sastre, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+recombinant+influenza+virus+from+plasmid+DNA.">Generation of recombinant influenza virus from plasmid DNA.</a> J Vis Exp. (42), (2010).</li><li>National Research Council (U.S.) Committee for the Update of the Guide for the Care and Use of Laboratory Animals., Institute for Laboratory Animal Research (U.S.). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the care and use of laboratory animals. , (2011).</li><li>Reed, L. J., Muench, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+of+estimating+fifty+percent+endpoints.">A simple method of estimating fifty percent endpoints.</a> The American Journal of Hygiene. 27 (3), 493-497 (1938).</li><li>Golde, W. T., Gollobin, P., Rodriguez, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rapid,+simple,+and+humane+method+for+submandibular+bleeding+of+mice+using+a+lancet.">A rapid, simple, and humane method for submandibular bleeding of mice using a lancet.</a> Lab Animal. 34 (9), 39-43 (2005).</li><li>Nogales, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+temperature+sensitive+live-attenuated+canine+influenza+virus+H3N8+vaccine.">A temperature sensitive live-attenuated canine influenza virus H3N8 vaccine.</a> J Virol. , (2016).</li><li>Eisfeld, A. J., Neumann, G., Kawaoka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influenza+A+virus+isolation,+culture+and+identification.">Influenza A virus isolation, culture and identification.</a> Nat Protoc. 9 (11), 2663-2681 (2014).</li><li>Guo, H., Baker, S. F., Martinez-Sobrido, L., Topham, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+CD8+T+cell+heterologous+protection+by+a+single+dose+of+single-cycle+infectious+influenza+virus.">Induction of CD8 T cell heterologous protection by a single dose of single-cycle infectious influenza virus.</a> J Virol. 88, 12006-12016 (2014).</li><li>He, W., Mullarkey, C. E., Miller, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+the+neutralization+potency+of+influenza+A+virus+hemagglutinin+stalk/stem-binding+antibodies+in+polyclonal+preparations+by+microneutralization+assay.">Measuring the neutralization potency of influenza A virus hemagglutinin stalk/stem-binding antibodies in polyclonal preparations by microneutralization assay.</a> Methods. , (2015).</li><li>Gulati, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibody+epitopes+on+the+neuraminidase+of+a+recent+H3N2+influenza+virus+(A/Memphis/31/98).">Antibody epitopes on the neuraminidase of a recent H3N2 influenza virus (A/Memphis/31/98).</a> J Virol. 76 (23), 12274-12280 (2002).</li><li>Beare, A. S., Webster, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Replication+of+avian+influenza+viruses+in+humans.">Replication of avian influenza viruses in humans.</a> Arch Virol. 119, 37-42 (1991).</li><li>Rowe, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+antibody+to+avian+influenza+A+(H5N1)+virus+in+human+serum+by+using+a+combination+of+serologic+assays.">Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays.</a> J Clin Microbiol. 37 (4), 937-943 (1999).</li><li>Stephenson, I., Wood, J. M., Nicholson, K. G., Zambon, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sialic+acid+receptor+specificity+on+erythrocytes+affects+detection+of+antibody+to+avian+influenza+haemagglutinin.">Sialic acid receptor specificity on erythrocytes affects detection of antibody to avian influenza haemagglutinin.</a> J Med Virol. 70 (3), 391-398 (2003).</li></ol>

Los autores no tienen nada que revelar.

El modelo de ratón de IAV es ampliamente utilizado en vivo estudios de eficacia IAV patogenesia, inmunogenicidad y protección. El pequeño tamaño de los ratones los hace fáciles de manipular y almacenar en comparación con otros modelos animales tales como hurones o cobayas. Además, la facilidad en términos de costo de animal, alojamiento y reproducción permiten su uso en pruebas de vacunación pre clínicos en los que se necesitan grandes cantidades de animales. En particular, ya que los ratones se han u...

Gripe es virus envueltos, clasificados en la familia de Orthomyxoviridae 3. Contienen ocho moléculas de ARN monocatenario con polaridad negativa3. En los seres humanos, gripe causa epidemias estacionales y pandemias ocasionales de consecuencia importante cuando se introducen nuevos virus en la población humana4. Por otra parte, gripe estacional es altamente y rápidamente transmitida entre los seres humanos produciendo una elevada pérdida económica en todo el mundo cada año2,5. IAV los síntomas incluyen tos, congestión nasal, fiebre, malestar general, cefalea, anorexia y mialgias, pero el virus también puede producir una enfermedad más severa en pacientes inmunocomprometidos6. De hecho, la Organización Mundial de la salud (OMS) calcula que el virus de la gripe estacional causan 300.000-500.000 muertes en todo el mundo cada año1. Hay sólo dos clases de medicamentos actualmente aprobados por la Food and Drug Administration (FDA) para la profilaxis de la gripe y tratamiento en seres humanos: inhibidores de la neuraminidasa (NA) (p. ej., oseltamivir) y bloqueadores del canal iónico M2 (p. ej., amantadina); sin embargo, la aparición de variantes del virus resistentes a los medicamentos es una preocupación cada vez mayor. Vacunación, por lo tanto, sigue siendo la mejor opción médica para proteger a los seres humanos contra las infecciones de gripe. Hasta la fecha, tres tipos de influenza hay vacunas aprobadas por la FDA para el uso humano: las vacunas de proteínas recombinantes virales hemagglutinin (HA), inactivadas vacunas contra la influenza (IIV) y viva atenuada de influenza vacunas (LAIV)5, 7. las tres vacunas están diseñadas para inducir la respuesta inmune adaptativa contra la proteína viral de la HA, el objetivo principal de neutralizar anticuerpos contra la gripe.
Un modelo de ratón validado para el estudio de infección de IAV en vivo
Se han utilizado modelos animales para estudiar, entre otros, IAV patogenesia8,9,10,11, factores virales que contribuyen a la enfermedad12 o transmisión viral13 ,14, y probar la eficacia de nuevas vacunas o antivirales fármacos9,10,15. Ratones (Mus musculus) son el modelo animal más ampliamente utilizado para la investigación IAV por varias razones: 1) el sistema inmune es evolutivamente similar al que presentan en los seres humanos; compra animal de bajo costo, incluyendo 2), la vivienda y la reproducción; 3) pequeño tamaño fácilmente manipular y almacenar; 4) variabilidad de host mínimo para obtener respuestas homogéneas y resultados; 5) un gran conocimiento de la biología de ratones, incluyendo la secuencia del genoma; 6) muchos disponible biología molecular y/o reactivos de Inmunología; 7) disponible knock out (KO) de ratones para estudiar la contribución de una proteína huésped dado en infección viral; y, 8) múltiples cepas de ratón que pueden ser aprovechadas para evaluar las bases genéticas de las infecciones.
Hay varias cepas de ratón disponibles actualmente para el estudio de IAV en vivo. Edad, estado inmunitario, sexo, cepa genética de fondo y el ratón, así como rutas de infección, dosis y cepas del virus de influyen en el resultado de la infección del IAV en ratones. Las cepas de ratón más comunes utilizadas en la investigación IAV son C57BL/6, BALB/C y, más recientemente, DBA.2 ratones ya que son más susceptibles a la enfermedad IAV que dos cepas anteriores16,17,18, 19 , 20. lo que es importante, la respuesta inmune también puede ser diferente dependiendo de la cepa de ratón18,19,20. Por lo tanto, es muy importante recuperar toda la información disponible sobre el ratón y la cepa IAV a elegir la mejor opción para el experimento a realizarse.
Aunque el ratón es un buen modelo animal de infección para los estudios en vivo IAV, tienen varias limitaciones, que deben considerarse en el diseño experimental. Por ejemplo, una limitación importante de la utilización de ratones en vivo estudios es que la gripe no transmite entre ratones. Así, para la transmisión de los estudios, más aceptado modelos animales (e.g., hurones o cobayas) son usados16,17,21. Además, hay varias diferencias entre las manifestaciones del IAV en ratones y seres humanos. A diferencia de los seres humanos, ratones no desarrollan fiebre con infección de IAV; por el contrario presentan hipotermia16,17. En ratones, replicación de IAV se concentra en el tracto respiratorio inferior (pulmones) en lugar de las vías aéreas superiores. Por lo tanto, virulencia del IAV en ratones no se correlaciona siempre al que se observa en los seres humanos. En conjunto, ya que las ventajas compensan las desventajas limitadas, ratón representa el primer modelo animal para evaluar patogenesia viral gripe, inmunogenicidad y eficacia protectora en los estudios de vacunas y antivirales. Por otra parte, no sería éticamente aceptable para llevar a cabo estudios con IAV usando modelos animales grandes sin pruebas previas en un pequeño modelo animal de infección IAV. En este manuscrito, describimos cómo infectar ratones intranasal (i.n.) con IAV, cómo controlar la severidad y progreso de la infección viral y cómo llevar a cabo los experimentos necesarios para evaluar la respuesta inmune humoral y la eficacia de protección.

<table><tbody><tr><td>Madin-Darby Canine Kidney (MDCK) epithelial cells</td><td>ATCC</td><td>CCL-34</td><td></td></tr><tr><td>Six- to eight-week-old female C57BL/6 mice</td><td>National Cancer Institute (NCI)</td><td>01XBE</td><td></td></tr><tr><td>Turckey red blod cells</td><td>Biolink Inc</td><td></td><td>Store at 4 &amp;deg;C</td></tr><tr><td>Dulbecco&amp;rsquo;s modified Eagle&amp;rsquo;s medium (DMEM)</td><td>Corning Cellgro</td><td>15-013-CV</td><td>Store at 4 &amp;deg;C</td></tr><tr><td>Fetal Bovine Serum (FBS)</td><td>Seradigm</td><td>1500-050</td><td>Store at -20 &amp;deg;C</td></tr><tr><td>Penicillin/Streptomycin/L-Glutamine (PSG) 100x</td><td>Corning</td><td>30-009-CI</td><td>Store at -20 &amp;deg;C</td></tr><tr><td>Penicillin/Streptomycin (PS) 100x</td><td>Corning</td><td>30-00-CI</td><td>Store at -20 &amp;deg;C</td></tr><tr><td>Bovin Albumin solution (BA)</td><td>Sigma-Aldrich</td><td>A7409</td><td>Store at 4 &amp;deg;C</td></tr><tr><td>Bovin Serum Albumin (BSA)</td><td>Sigma-Aldrich</td><td>A9647</td><td>Store at 4 &amp;deg;C</td></tr><tr><td>Tosylsulfonyl phenylalanyl chloromethyl ketone (TPCK)-treated trypsin</td><td>Sigma-Aldrich</td><td>T8802</td><td>Store at -20 &amp;deg;C</td></tr><tr><td>Neutral Buffered Formalin 10%</td><td>EMD</td><td>65346-85</td><td>Store at RT</td></tr><tr><td>Triton X-100</td><td>J.T.Baker</td><td>X198-07</td><td>Store at RT</td></tr><tr><td>Monoclonal Antibody anti-NP Influenza A Virus HB-65</td><td>ATTC</td><td>H16-L10-4R5</td><td>Store at -20 &amp;deg;C</td></tr><tr><td>Polyclonal rabbit anti-mouse immunoglobulins/FITC</td><td>Dako</td><td>F0261</td><td>Store at 4 &amp;deg;C</td></tr><tr><td>ECL Anti-mouse IgG, Horseradish Peroxidase linked whole antibody</td><td>GE Healthcare</td><td>LNA931V/AG</td><td>Store at 4 &amp;deg;C</td></tr><tr><td>TMB substrate set</td><td>BioLegend</td><td>421101</td><td>Store at 4 &amp;deg;C</td></tr><tr><td>Vmax Kinetic plate reader</td><td>Molecular Devices</td><td></td><td></td></tr><tr><td>Dounce Tissue Grinders</td><td>Thomas Scientific</td><td>7722-7</td><td></td></tr><tr><td>Receptor destroying enzyme, RDE (II)</td><td>Denka Seiken Co.</td><td>370013</td><td>Store at -20 &amp;deg;C</td></tr><tr><td>Crystal Violet</td><td>Fisher Scienctific</td><td>C581-100</td><td>Store at RT</td></tr><tr><td>96-well Cell Culture Plate</td><td>Greiner Bio-one</td><td>655-180</td><td></td></tr><tr><td>Cell Culture dishes 100 mm</td><td>Greiner Bio-one</td><td>664-160</td><td></td></tr><tr><td>Nunc MicroWell 96-Well Microplates</td><td>Thermo Fisher Scienctific</td><td>269620</td><td></td></tr><tr><td>Nunc 96-Well Polystyrene Conical Bottom MicroWell Plates</td><td>Thermo Fisher Scienctific</td><td>249570</td><td></td></tr><tr><td>Puralub Vet Ointment</td><td>Dechra</td><td>9N-76855</td><td></td></tr><tr><td>Fluorescent microscope</td><td>Olympus</td><td>Olympus IX81</td><td></td></tr></tbody></table>

influenza a virus studies in a mouse model of infection

Virus de la influenza causan más de 500.000 muertes en todo el mundo1 y se asocian con un costo anual de US $ 12 billones en los Estados Unidos solo considerando médica directa y gastos de hospitalización y ausentismo de trabajo2. Modelos animales son cruciales en los estudios de Influenza A virus (IAV) evaluar patogenesia viral, interacciones huésped-patógeno, respuesta inmune, y la eficacia de la vacuna actual o nuevos enfoques así como antivirales. Los ratones son un modelo animal pequeño ventajoso porque su sistema inmunológico es evolutivamente similar a ése encontrado en los seres humanos, están disponibles de proveedores comerciales como sujetos genéticamente idénticos, existen múltiples cepas que pueden ser aprovechadas para evaluar la base genética de las infecciones, y son relativamente baratas y fáciles de manipular. Para recapitular la infección IAV en los seres humanos a través de las vías respiratorias, ratones primero son anestesiados antes de la inoculación intranasal con gripe infecciosa bajo contención de bioseguridad adecuado. Después de la infección, la patogenesia de la gripe se determina mediante el control diario de la morbilidad (pérdida de peso corporal) y la tasa de mortalidad (supervivencia). Además, patogenesia viral también puede evaluarse mediante la evaluación de replicación del virus en la parte superior (mucosa nasal) o tracto respiratorio (pulmones) inferior de ratones infectados. Las respuestas humorales sobre infección de IAV pueden rápidamente evaluar sangrado no invasiva y la detección del anticuerpo secundario ensayos destinados a detectar la presencia de total o anticuerpos neutralizantes. Aquí, describimos los métodos comunes utilizados para infectar ratones intranasal (i.n) con IAV y evaluar patogénesis, respuesta inmune humoral y eficacia de protección.

Caracterización de la patogenesia viral en ratones
La patogenesia de IAV se relaciona con la morbilidad y la mortalidad causada por la infección. Estos dos parámetros pueden evaluarse fácilmente en ratones: morbilidad IAV se asocia con pérdida de peso corporal en ratones infectados y el porcentaje de supervivencia indicará la tasa de mortalidad (figura 1). El peso corporal y la supervivencia en r...

Watch this Scientific Journal Video about Influenza A Virus Studies in a Mouse Model of Infection at JoVE.com

Estudios del virus de la influenza A en un modelo de infección en ratones | Inmunología e Infección | Video

Influenza A Virus Studies in a Mouse Model of Infection

Virus de la gripe A (gripe) son importantes patógenos respiratorios humanos. Para entender la patogenicidad de gripe y realizar ensayos preclínicos de vacunas nuevos enfoques, modelos animales, imitar la fisiología humana se requieren. Aquí, describimos las técnicas para evaluar el IAV patogenia, respuesta humoral y eficacia de la vacuna utilizando un modelo murino de infección.

Estudios de Virus de influenza A en un modelo murino de infección

Influenza viruses cause over 500,000 deaths worldwide1 and are associated with an annual cost of 12 - 14 billion USD in the United States alone ...

influenza-a-virus-studies-in-a-mouse-model-of-infection

Research

Education

JoVE Journal

JoVE Core

Biology

Immunology and Infection

Genetics

Viruses

SCIENTISTS IN ACTION

28.2K vistas.  University of Rochester School of Medicine and Dentistry. Virus de la gripe A (gripe) son importantes patógenos respiratorios humanos. Para entender la patogenicidad de gripe y realizar ensayos preclínicos de vacunas nuevos enfoques, modelos animales, imitar la fisiología humana se requieren. Aquí, describimos las técnicas para evaluar el IAV patogenia, respuesta humoral y eficacia de la vacuna utilizando un modelo murino de infección.

Video: Influenza A Virus Studies in a Mouse Model of Infection

Using Bioluminescent Imaging to Investigate Synergism Between Streptococcus pneumoniae and Influenza A Virus in Infant Mice

During the 1918 influenza virus pandemic, which killed approximately 50 million people worldwide, the majority of fatalities were not the result of infection with influenza virus alone. Instead, most individuals are thought to have succumbed to a secondary bacterial infection, predominately caused by the bacterium Streptococcus pneumoniae (the pneumococcus). The synergistic relationship between infections caused by influenza virus and the pneumococcus has subsequently been observed during the 1957 Asian influenza virus pandemic, as well as during seasonal outbreaks of the virus (reviewed in 1, 2). Here, we describe a protocol used to investigate the mechanism(s) that may be involved in increased morbidity as a result of concurrent influenza A virus and S. pneumoniae infection. We have developed an infant murine model to reliably and reproducibly demonstrate the effects of influenza virus infection of mice colonised with S. pneumoniae. Using this protocol, we have provided the first insight into the kinetics of pneumococcal transmission between co-housed, neonatal mice using in vivo imaging 3.

Using Bioluminescent Imaging to Investigate Synergism Between Streptococcus pneumoniae and Influenza A Virus in Infant Mice

A concurrent infection with influenza A virus is one of the factors implicated in the induction of invasive pneumococcal disease during asymptomatic Streptococcus pneumoniae carriage. Here we describe a mixed infection method using infant mice to investigate the synergism between these two respiratory pathogens.

Utilizando imágenes bioluminiscentes encargado de investigar la sinergia entre Streptococcus pneumoniae Y el Virus de la Influenza en ratones lactantes

During the 1918 influenza virus pandemic, which killed approximately 50 million people worldwide, the majority of fatalities were not the result of ...

Investigación

Inmunología e Infección

High-throughput Detection Method for Influenza Virus

Influenza virus is a respiratory pathogen that causes a high degree of morbidity and mortality every year in multiple parts of the world. Therefore, precise diagnosis of the infecting strain and rapid high-throughput screening of vast numbers of clinical samples is paramount to control the spread of pandemic infections. Current clinical diagnoses of influenza infections are based on serologic testing, polymerase chain reaction, direct specimen immunofluorescence and cell culture 1,2.
Here, we report the development of a novel diagnostic technique used to detect live influenza viruses. We used the mouse-adapted human A/PR/8/34 (PR8, H1N1) virus 3 to test the efficacy of this technique using MDCK cells 4. MDCK cells (104 or 5 x 103 per well) were cultured in 96- or 384-well plates, infected with PR8 and viral proteins were detected using anti-M2 followed by an IR dye-conjugated secondary antibody. M2 5 and hemagglutinin 1 are two major marker proteins used in many different diagnostic assays. Employing IR-dye-conjugated secondary antibodies minimized the autofluorescence associated with other fluorescent dyes. The use of anti-M2 antibody allowed us to use the antigen-specific fluorescence intensity as a direct metric of viral quantity. To enumerate the fluorescence intensity, we used the LI-COR Odyssey-based IR scanner. This system uses two channel laser-based IR detections to identify fluorophores and differentiate them from background noise. The first channel excites at 680 nm and emits at 700 nm to help quantify the background. The second channel detects fluorophores that excite at 780 nm and emit at 800 nm. Scanning of PR8-infected MDCK cells in the IR scanner indicated a viral titer-dependent bright fluorescence. A positive correlation of fluorescence intensity to virus titer starting from 102-105 PFU could be consistently observed. Minimal but detectable positivity consistently seen with 102-103 PFU PR8 viral titers demonstrated the high sensitivity of the near-IR dyes. The signal-to-noise ratio was determined by comparing the mock-infected or isotype antibody-treated MDCK cells.
Using the fluorescence intensities from 96- or 384-well plate formats, we constructed standard titration curves. In these calculations, the first variable is the viral titer while the second variable is the fluorescence intensity. Therefore, we used the exponential distribution to generate a curve-fit to determine the polynomial relationship between the viral titers and fluorescence intensities. Collectively, we conclude that IR dye-based protein detection system can help diagnose infecting viral strains and precisely enumerate the titer of the infecting pathogens.

This method describes the use of Infrared dye based imaging system for detection of H1N1 in bronchioalveolar lavage (BAL) fluid of infected mice at a high sensitivity. This methodology can be performed in a 96- or 384-well plate, requires &lt;10 &mu;l volume of test material and has the potential for concurrent screening of multiple pathogens.

De alto rendimiento Método de detección de virus de la influenza

Influenza virus is a respiratory pathogen that causes a high degree of morbidity and mortality every year in multiple parts of the world. Therefore, ...

Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity

Vaccines are one of the greatest achievements of mankind, and have saved millions of lives over the last century. Paradoxically, little is known about the physiological mechanisms that mediate immune responses to vaccines perhaps due to the overall success of vaccination, which has reduced interest into the molecular and physiological mechanisms of vaccine immunity. However, several important human pathogens including influenza virus still pose a challenge for vaccination, and may benefit from immune-based strategies. 
Although influenza reverse genetics has been successfully applied to the generation of live-attenuated influenza vaccines (LAIVs), the addition of molecular tools in vaccine preparations such as tracer components to follow up the kinetics of vaccination in vivo, has not been addressed. In addition, the recent generation of mouse models that allow specific depletion of leukocytes during kinetic studies has opened a window of opportunity to understand the basic immune mechanisms underlying vaccine-elicited protection. Here, we describe how the combination of reverse genetics and chimeric mouse models may help to provide new insights into how vaccines work at physiological and molecular levels, using as example a recombinant, cold-adapted, live-attenuated influenza vaccine (LAIV). We utilized laboratory-generated LAIVs harboring cell tracers as well as competitive bone marrow chimeras (BMCs) to determine the early kinetics of vaccine immunity and the main physiological mechanisms responsible for the initiation of vaccine-specific adaptive immunity. In addition, we show how this technique may facilitate gene function studies in single animals during immune responses to vaccines. We propose that this technique can be applied to improve current prophylactic strategies against pathogens for which urgent medical countermeasures are needed, for example influenza, HIV, Plasmodium, and hemorrhagic fever viruses such as Ebola virus.

There is an overall lack of knowledge about how vaccines work. Here we propose the combined use of reverse genetics and bone marrow chimeric mice to gain insight into the early host immune responses to vaccines with a special focus on dendritic cells and T cell immunity.

Administración intranasal de vacunas contra la influenza recombinante en modelos de ratón quimérico para el Estudio de la mucosa Inmunidad

Vaccines are one of the greatest achievements of mankind, and have saved millions of lives over the last century. Paradoxically, little is known about the ...

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Studying the interaction between proteins is key in understanding their function(s). A very powerful method that is frequently used to study interactions of proteins with other macromolecules in a complex sample is called co-immunoprecipitation. The described co-immunoprecipitation protocol allows to demonstrate and further investigate the interaction between the antiviral myxovirus resistance protein 1 (Mx1) and one of its viral targets, the influenza A virus nucleoprotein (NP). The protocol starts with transfected mammalian cells, but it is also possible to use influenza A virus infected cells as starting material. After cell lysis, the viral NP protein is pulled-down with a specific antibody and the resulting immune-complexes are precipitated with protein G beads. The successful pull-down of NP and the co-immunoprecipitation of the antiviral Mx1 protein are subsequently revealed by western blotting. A prerequisite for successful co-immunoprecipitation of Mx1 with NP is the presence of N-ethylmaleimide (NEM) in the cell lysis buffer. NEM alkylates free thiol groups. Presumably this reaction stabilizes the weak and/or transient NP&#8211;Mx1 interaction by preserving a specific conformation of Mx1, its viral target or an unknown third component. An important limitation of co-immunoprecipitation experiments is the inadvertent pull-down of contaminating proteins, caused by nonspecific binding of proteins to the protein G beads or antibodies. Therefore, it is very important to include control settings to exclude false positive results. The described co-immunoprecipitation protocol can be used to study the interaction of Mx proteins from different vertebrate species with viral proteins, any pair of proteins, or of a protein with other macromolecules. The beneficial role of NEM to stabilize weak and/or transient interactions needs to be tested for each interaction pair individually.

This co-immunoprecipitation protocol allows to study the interaction between the influenza A virus nucleoprotein and the antiviral Mx1 protein in human cells. The protocol emphasizes the importance of N-ethylmaleimide for successful co-immunoprecipitation of Mx1 and influenza A virus nucleoprotein.

Co-inmunoprecipitación de la proteína de ratón Mx1 con la Influenza A nucleoproteína del virus

Studying the interaction between proteins is key in understanding their function(s). A very powerful method that is frequently used to study interactions ...

Using Zebrafish Models of Human Influenza A Virus Infections to Screen Antiviral Drugs and Characterize Host Immune Cell Responses

Each year, seasonal influenza outbreaks profoundly affect societies worldwide. In spite of global efforts, influenza remains an intractable healthcare burden. The principle strategy to curtail infections is yearly vaccination. In individuals who have contracted influenza, antiviral drugs can mitigate symptoms. There is a clear and unmet need to develop alternative strategies to combat influenza. Several animal models have been created to model host-influenza interactions. Here, protocols for generating zebrafish models for systemic and localized human influenza A virus (IAV) infection are described. Using a systemic IAV infection model, small molecules with potential antiviral activity can be screened. As a proof-of-principle, a protocol that demonstrates the efficacy of the antiviral drug Zanamivir in IAV-infected zebrafish is described. It shows how disease phenotypes can be quantified to score the relative efficacy of potential antivirals in IAV-infected zebrafish. In recent years, there has been increased appreciation for the critical role neutrophils play in the human host response to influenza infection. The zebrafish has proven to be an indispensable model for the study of neutrophil biology, with direct impacts on human medicine. A protocol to generate a localized IAV infection in the Tg(mpx:mCherry) zebrafish line to study neutrophil biology in the context of a localized viral infection is described. Neutrophil recruitment to localized infection sites provides an additional quantifiable phenotype for assessing experimental manipulations that may have therapeutic applications. Both zebrafish protocols described faithfully recapitulate aspects of human IAV infection. The zebrafish model possesses numerous inherent advantages, including high fecundity, optical clarity, amenability to drug screening, and availability of transgenic lines, including those in which immune cells such as neutrophils are labeled with fluorescent proteins. The protocols detailed here exploit these advantages and have the potential to reveal critical insights into host-IAV interactions that may ultimately translate into the clinic.

Systemic and localized zebrafish infection models for human influenza A virus are demonstrated. Using a systemic infection model, zebrafish can be used to screen antiviral drugs. Using a localized infection model, zebrafish can be used to characterize host immune cell responses.

El uso de modelos de pez cebra de Infecciones por Virus de la Influenza Humana A a la pantalla medicamentos antivirales y caracterizar inmune del huésped respuestas de las células

Each year, seasonal influenza outbreaks profoundly affect societies worldwide. In spite of global efforts, influenza remains an intractable healthcare ...

Imaging Cell Interaction in Tracheal Mucosa During Influenza Virus Infection Using Two-photon Intravital Microscopy

The analysis of cell-cell or cell-pathogen interaction in vivo is an important tool to understand the dynamics of the immune response to infection. Two-photon intravital microscopy (2P-IVM) allows the observation of cell interactions in deep tissue in living animals, while minimizing the photobleaching generated during image acquisition. To date, different models for 2P-IVM of lymphoid and non-lymphoid organs have been described. However, imaging of respiratory organs remains a challenge due to the movement associated with the breathing cycle of the animal.
Here, we describe a protocol to visualize in vivo immune cell interactions in the trachea of mice infected with influenza virus using 2P-IVM. To this purpose, we developed a custom imaging platform, which included the surgical exposure and intubation of the trachea, followed by the acquisition of dynamic images of neutrophils and dendritic cells (DC) in the mucosal epithelium. Additionally, we detailed the steps needed to perform influenza intranasal infection and flow cytometric analysis of immune cells in the trachea. Finally, we analyzed neutrophil and DC motility as well as their interactions during the course of a movie. This protocol allows for the generation of stable and bright 4D images necessary for the assessment of cell-cell interactions in the trachea.

In this study, we present a protocol to perform two-photon intravital imaging and cell interaction analysis in the murine tracheal mucosa after infection with influenza virus. This protocol will be relevant for researchers studying immune cell dynamics during respiratory infections.

Proyección de imagen de interacción de células en Mucosa traqueal durante la infección del Virus de la Influenza usando microscopia Intravital de dos fotones

The analysis of cell-cell or cell-pathogen interaction in vivo is an important tool to understand the dynamics of the immune response to infection. ...

A Murine Model of Dengue Virus-induced Acute Viral Encephalitis-like Disease

Dengue virus (DENV), an arthropod-borne virus transmitted by mosquitoes, may cause the severe disease known as dengue hemorrhagic fever, which is characterized by lethal complications due to plasma leakage, ascites, pleural effusion, respiratory distress, severe bleeding, and organ impairment. A few cases of DENV infection present neurological manifestations; however, studies have not explored DENV-induced neuropathogenesis further. In this study, we present a protocol to use an immunocompetent outbred ICR (Institute of Cancer Research) mouse for investigating the induction of central nervous system (CNS) infection with DENV, followed by the progression of acute viral encephalitis-like disease.

Here, we present a protocol for creating an immunocompetent ICR (Institute of Cancer Research) murine model of central nervous system infection to display the development of neuropathy. Monitoring acute viral encephalitic disorders by identical disease scores could be performed for showing dengue virus-induced neuropathy in vivo.

Un modelo murino de enfermedad similar a la encefalitis vírica aguda inducida por el virus del dengue

Dengue virus (DENV), an arthropod-borne virus transmitted by mosquitoes, may cause the severe disease known as dengue hemorrhagic fever, which is ...

A Luciferase-fluorescent Reporter Influenza Virus for Live Imaging and Quantification of Viral Infection

Influenza A viruses (IAVs) cause human respiratory disease that is associated with significant health and economic consequences. As with other viruses, studying IAV requires the use of laborious secondary approaches to detect the presence of the virus in infected cells and/or in animal models of infection. This limitation has been recently circumvented with the generation of recombinant IAVs expressing easily traceable fluorescent or bioluminescent (luciferase) reporter proteins. However, researchers have been forced to select fluorescent or luciferase reporter genes due to the restricted capacity of the IAV genome for including foreign sequences. To overcome this limitation, we have generated a recombinant replication-competent bi-reporter IAV (BIRFLU) stably expressing both a fluorescent and a luciferase reporter gene to easily track IAV infections in vitro and in vivo. To this end, the viral non-structural (NS) and hemagglutinin (HA) viral segments of influenza A/Puerto Rico/8/34 H1N1 (PR8) were modified to encode the fluorescent Venus and the bioluminescent Nanoluc luciferase proteins, respectively. Here, we describe the use of BIRFLU in a mouse model of IAV infection and the detection of both reporter genes using an in vivo imaging system. Notably, we have observed a good correlation between the expressions of both reporters and viral replication. The combination of cutting-edge techniques in molecular biology, animal research and imaging technologies, provides researchers the unique opportunity to use this tool for influenza research, including the study of virus-host interactions and dynamics of viral infections. Importantly, the feasibility to genetically alter the viral genome to express two foreign genes from different viral segments opens up opportunities to use this approach for: (i) the development of novel IAV vaccines, (ii) the generation of recombinant IAVs that can be used as vaccine vectors for the treatment of other human pathogen infections.

Influenza A viruses (IAVs) are contagious respiratory pathogens that cause annual epidemics and occasional pandemics. Here, we describe a protocol to track viral infections in vivo using a novel recombinant luciferase and fluorescence-expressing bi-reporter IAV (BIRFLU). This approach provides researchers with an excellent tool to study IAV in vivo.

Un virus de la gripe Reportero fluorescente de Luciferase para imágenes en vivo y cuantificación de la infección viral

Influenza A viruses (IAVs) cause human respiratory disease that is associated with significant health and economic consequences. As with other viruses, ...

Evaluation of T Follicular Helper Cells and Germinal Center Response During Influenza A Virus Infection in Mice

T Follicular Helper (Tfh) cells are an independent CD4+&#160;T cell subset specialized in providing help for germinal center (GC) development and generation of high-affinity antibodies. In influenza virus infection, robust Tfh and GC B cell responses are induced to facilitate effective virus eradication, which confers a qualified mouse model for Tfh-associated study. In this paper, we described protocols in detection of basic Tfh-associated immune response during influenza virus infection in mice. These protocols include: intranasal inoculation of influenza virus; flow cytometry staining and analysis of polyclonal and antigen-specific Tfh cells, GC B cells and plasma cells; immunofluorescence detection of GCs; enzyme-linked immunosorbent assay (ELISA) of influenza virus-specific antibody in serum. These assays basically quantify the differentiation and function of Tfh cells in influenza virus infection, thus providing help for studies in elucidating differentiation mechanism and manipulation strategy.

This paper describes protocols of evaluating Tfh and GC B response in mouse model of influenza virus infection.

Evaluación de las células auxiliares foliculares T y la respuesta del centro germinal durante la infección por el virus de la gripe A en ratones

T Follicular Helper (Tfh) cells are an independent CD4+&#160;T cell subset specialized in providing help for germinal center (GC) development and ...

A Mouse Model for the Transition of Streptococcus pneumoniae from Colonizer to Pathogen upon Viral Co-Infection Recapitulates Age-Exacerbated Illness

Streptococcus pneumoniae (pneumococcus) is an asymptomatic colonizer of the nasopharynx in most individuals but can progress to a pulmonary and systemic pathogen upon influenza A virus (IAV) infection. Advanced age enhances host susceptibility to secondary pneumococcal pneumonia and is associated with worsened disease outcomes. The host factors driving those processes are not well defined, in part due to a lack of animal models that reproduce the transition from asymptomatic colonization to severe clinical disease. 
This paper describes a novel mouse model that recreates the transition of pneumococci from asymptomatic carriage to disease upon viral infection. In this model, mice are first intranasally inoculated with biofilm-grown pneumococci to establish asymptomatic carriage, followed by IAV infection of both the nasopharynx and lungs. This results in bacterial dissemination to the lungs, pulmonary inflammation, and obvious signs of illness that can progress to lethality. The degree of disease is dependent on the bacterial strain and host factors. 
Importantly, this model reproduces the susceptibility of aging, because compared to young mice, old mice display more severe clinical illness and succumb to disease more frequently. By separating carriage and disease into distinct steps and providing the opportunity to analyze the genetic variants of both the pathogen and the host, this S. pneumoniae/IAV co-infection model permits the detailed examination of the interactions of an important pathobiont with the host at different phases of disease progression. This model can also serve as an important tool for identifying potential therapeutic targets against secondary pneumococcal pneumonia in susceptible hosts.

A Mouse Model for the Transition of Streptococcus pneumoniae from Colonizer to Pathogen upon Viral Co-Infection Recapitulates Age-Exacerbated Illness

This paper describes a novel mouse model for the transition of pneumococcus from an asymptomatic colonizer to a disease-causing pathogen during viral infection. This model can be readily adapted to study polymicrobial and host-pathogen interactions during the different phases of disease progression and across various hosts.

Un modelo de ratón para la transición de Streptococcus pneumoniae de colonizador a patógeno tras la coinfección viral recapitula la enfermedad exacerbada por la edad

Streptococcus pneumoniae (pneumococcus) is an asymptomatic colonizer of the nasopharynx in most individuals but can progress to a pulmonary and systemic ...

Estudios de Virus de influenza A en un modelo murino de infección

Resumen

Explorar más videos

PLAYLIST