rologie i - European Congress of Virology

5 th European Congress of VirologyThursday 12 th September 2013, 8h30 – 10h30WORKSHOP 25: “RESPIRATORY VIRUS INFECTIONS”KEYNOTE:Chairpersons: Fausto BALDANTI (Pavia, ITALY)& Astrid VABRET (Caen, FRANCE)AmphitheaterRespiratory virus infections: from pathogenesis to pandemicsPeter OPENSHAWProfessor of Experimental Medicine, Imperial College London W2 1PG,UNITED KINGDOMDespite concerns over population growth, water shortages, global warming,political and military strife, pandemic diseases remain major threatsto global prosperity and stability. In addition, health and wellbeing areconstantly eroded by common colds which act as co-factors in manycommon diseases.Our understanding the mechanism of disease induction by respiratorypathogens has undergone radical change over the past two decades. Respiratorysyncytial virus used to be thought of a cause of serious diseaseonly in infants but is increasingly recognized as a cause of morbidity andmortality in the elderly and immunocompromised. Natural infections aredifficult to study and animal models have limitations.Human experimental challenge is uniquely able to provide detailedinsights into the mechanisms of disease in the natural host. We have infectedadult volunteers with Memphis 37 strain of RSV, causing reproducibledisease without serious adverse events. We have found that viral load isrelated to and severity of symptoms, strengthening the case for the developmentof antiviral drugs. Importantly, there we found only an indirect andweak association between baseline antiviral antibody and susceptibilityto viral challenge. These studies need to be complemented by observationalstudies of natural infections, but are providing novel insights that areleading to improved understanding of viral disease.Studies of individual responses to viral infection are also helpful in planningfor outbreaks of respiratory disease. The UK made detailed pandemicplans in response to avian influenza; these plans were tested by the 2009-10 influenza pandemic. Although generally mild, there were over 8,000hospitalizations in the UK and probably some 250,000 deaths worldwide(to August 2010). The disease was especially severe in some patients withasthma, in pregnant women and in children under 5 years of age. However,many of those admitted to hospital had no known risk factors and wereotherwise young and healthy.We established two research consortia to study disease in UK hospitals: aclinical information network based on case audit (Flu-CIN) and an in-depthstudy of the pathogenesis of influenza with intensive collection and analysisof samples from the respiratory tract and periphery during diseaseand recovery. This second study (the Mechanisms Of Severe InfluenzaConsortium, MOSAIC) involved 45 co-investigators and recruited 255patients with influenza-like illness from 11 hospitals (total 4800 beds).Of these, 170 patients had confirmed influenza (87% pH1N1); the causeof illness was determined in all cases. Rooted in extensive clinical information(>4000 data fields per patient), various research teams focused onhost (cellular immunology, soluble mediator responses, transcriptomicsand genomics), pathogen (influenza virus genetic and antigenic characteristics,viral shedding and viral load) and co-pathogen (co-infecting orsecondarily-infecting bacteria and viruses), with the aim of explaining thevariable response to pH1N1 infection.This unique study is allowing a series of related research questions to beexamined, linking the information on a case-by-case basis from patientsand controls. The consortium has identified a human gene deletion (witha mouse homologue) linked to defective host responses in about 6% ofour cases but is rarely present in normal individuals. This has been confirmedon other populations in the Far East. The success of this consortiumdemonstrates that it is possible to conduct high-quality research during anunexpected pandemic, but emphasizes the need to maintain infrastructureand preparedness.ORAL COMMUNICATIONSREF O43Growth and characterization of different HRV C types in 3D humanairway epithelia reconstituted in vitroKomla SOBO 1,2 , Caroline TAPPAREL 1,2 , Laurent KAISER 1,2 , SandraVAN BELLE 2 , Samuel CONSTANT 3 , Song HUANG 31 University of Geneva, Geneva, SWITZERLAND; 2 Laboratory of Virology,Division of Infectious Diseases and Division of Laboratory Medicine, Universityof Geneva Hospitals, Geneva, SWITZERLAND; 3 Epithelix sárl, 14Chemin des Aulx, 1228 Plan les Ouates, Geneva, Switzerland, Geneva,SWITZERLANDThe development of new molecular diagnostic tools has recently allowedthe discovery of human rhinovirus species C (HRV C). Several reportssuggest that HRV Cs may be overrepresented in children with pneumoniaor acute wheezing and exacerbation of asthma. HRV Cs cannot be propagatedin conventional immortalized cell lines, hence unlike HRV As andHRV Bs, the biological properties of these species C rhinoviruses havebeen difficult to study. However three groups have recently described thesuccessful amplification of HRV C15, untyped HRV C (W23), HRV C11and HRV C41 in sinus mucosal organ cultures {Bochkov, 2011 #1009}and in fully differentiated human airways epithelial cells (nasal or bronchialepithelial) {Hao, 2012 #1006; Ashraf, 2012 #1010}. Consistent withthese recent works, we report here that a panel of clinical HRV C specimensincluding HRV C2, HRV C7, HRV C12, HRV C15 and HRV C29types were all capable of mediating productive infection in reconstituted3D human primary upper airway epithelial tissues and that the virionsenter and exit preferentially through the apical surface. Similar to HRVA and HRV B, our data furthermore confirm the acid sensitivity of HRVCs. Finally, we show that the optimum temperature requirement duringthe growth of HRV Cs may be type dependant.REF O44Replication of Human Rhinovirus C in a Standard Cell LineAnna GÖRANSSON 1 , Nina JONSSON 1 , Fredrik LYSHOLM 2 , TobiasALLANDER 2 , A Michael LINDBERG 1 , Björn ANDERSSON 21 Linnaeus University, Kalmar, SWEDEN; 2 Karolinska Institutet, Stockholm,SWEDENHuman rhinovirus C (HRV C), a recently discovered picornavirus, is frequentlydetected during lower respiratory tract infections, primarily inchildren. HRV C is often associated with more severe infections than thoseof HRV A and –B. Studies of HRV C has so far been hampered due to theabsence of simple and rapid methods to propagate this virus in standard celllines. Recently Hao et al. reported replicating HRV C after transfection offully differentiated airway epithelial cells with RNA derived from a HRVC cDNA clone. Although this is a step forward it requires the access toprimary epithelial cells. Here we present data that support the replicationof a strain of HRV C in the standard cell line; human rhabdomyosarcoma(RD) cells. In vitro transcribed viral RNA from a cDNA clone of HRV C34was transfected into RD cells and viral replication was verified by methodsdetecting accumulation of viral genomes. Further characterization of thereplicating HRV C34 strain will be presented.Hao W, Bernard K, Patel N, Ulbrandt N, Feng H, Svabek C, Wilson S,Stracener C, Wang K, Suzich J, Blair W, Zhu Q. Infection and Propagationof Human Rhinovirus C in Human Airway Epithelial Cells. J Virol 2012;86 (24): 13524-32.S54 Virologie, Vol 17, supplément 2, septembre 2013