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Exploratory Spatial Analysis of in vitro Respiratory Syncytial Virus Co-infections.doc

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Exploratory Spatial Analysis of in vitro Respiratory Syncytial Virus Co-infections

Viruses2010,2,2782-2802;doi:10.3390/v2122782 OPENACCESS viruses ISSN1999-4915 /journal/viruses Article ExploratorySpatialAnalysisofinvitroRespiratorySyncytial VirusCo-infections IvanSimeonov 1,?,XiaoyanGong 2,?,OekyungKim 2,MaryPoss 2,3,FrancescaChiaromonte 1 andJohnFricks 1,? 1 2 3 DepartmentofStatistics,PennsylvaniaStateUniversity,UniversityPark,PA16802,USA DepartmentofBiology,PennsylvaniaStateUniversity,UniversityPark,PA16802,USA FogartyInternationalCenter,NationalInstitutesofHealth,Bethesda,MD20892,USA Authortowhomcorrespondenceshouldbeaddressed;E-Mail:fricks@; Tel.:+1-814-865-3235;Fax:+1-814-863-7114. ? Theseauthorscontributedequallytothispaper. Received:15November2010;inrevisedform:12December2010/Accepted:13December2010/ Published:22December2010 Abstract: The cell response to virus infection and virus perturbation of that response is dynamic and is re?ected by changes in cell susceptibility to infection. In this study, we evaluated the response of human epithelial cells to sequential infections with human respiratory syncytial virus strains A2 and B to determine if a primary infection with one strainwillimpacttheabilityofcellstobeinfectedwiththesecondasafunctionofvirus strain and time elapsed between the two exposures. Infected cells were visualized with ?uorescentmarkers,andlocationofallcellsinthetissueculturewellwereidenti?edusing imaging software. We employed tools from spatial statistics to investigate the likelihood ofacellbeinginfectedgivenitsproximitytoacellinfectedwitheitherthehomologous or heterologous virus. We used point processes, K-functions, and simulation procedures designed to account for speci?c features of our data when assessing spatial associations. Ourresultssuggestthatintrinsiccellpropertiesincreasesusceptibilityofcellstoinfection, more so for RSV-B than for RSV-A. Further, we provide evidence that the primary infectioncandecreasesusceptibilityofcellstotheheterologouschallengevirusbutonlyat the 16 h time point evaluated in this