title: Induction of IL-6 and CCL5 (RANTES) in human respiratory epithelial (A549) cells by clinical isolates of respiratory syncytial virus is strain specific.
abstract: BACKGROUND
  Respiratory syncytial virus (RSV) is the major respiratory pathogen of infants and young children. During each seasonal epidemic, multiple strains of both subgroup A and B viruses circulate in the community. Like other RNA viruses, RSV genome replication is prone to errors that results in a heterogeneous population of viral strains some of which may possess differences in virulence. We sought to determine whether clinical isolates of RSV differ in their capacity to induce inflammatory cytokines IL-6 and CCL5 (previously known as RANTES [regulated upon activation, normal T-cell expressed and secreted protein]), which are known to be induced in vitro and in vivo in response to RSV, during infection of A549 cells.
  
  
  RESULTS
  Screening of subgroup A and B isolates revealed heterogeneity among strains to induce IL-6 and CCL5. We chose two subgroup B strains, New Haven (NH)1067 and NH1125, for further analysis because of their marked differences in cytokine inducing properties and because subgroup B strains, in general, are less genetically heterogeneous as compared to subgroup A strains. At 12 and 24 hours post infection RSV strains, NH1067 and NH1125 differed in their capacity to induce IL-6 by an order of magnitude or more. The concentrations of IL-6 and CCL5 were dependent on the dose of infectious virus and the concentration of these cytokines induced by NH1125 was greater than that of those induced by NH1067 when the multiplicity of infection of NH1067 used was as much as 10-fold higher than that of NH1125. The induction of IL-6 was dependent on viable virus as infection with UV-inactivated virus did not induce IL-6. The difference in IL-6 induction most likely could not be explained by differences in viral replication kinetics. The intracellular level of RSV RNA, as determined by quantitative RT-PCR, was indistinguishable between the 2 strains though the titer of progeny virus produced by NH1125 was greater than that produced by NH1067 at 16, 24 and 36 hours but essentially equal at 48 and 72 hours. Full genome sequencing of the 2 strains revealed 193 polymorphisms and 4 insertions in NH1067 when compared to NH1125 (2 single base insertions in non-coding regions and 2 duplications of 3 and 60 bases in the RSV G gene). Of the polymorphisms, 147 occurred in coding regions and only 30 resulted in amino acid changes in 7 of the RSV genes.
  
  
  CONCLUSIONS
  These data suggest that RSV strains may not be homogeneous with regard to pathogenesis or virulence. Identification of the genetic polymorphisms associated with variations in cytokine induction may lead to insights into RSV disease and to the development of effective antiviral agents and vaccines.
authors.affiliation: Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, TX, USA.
authors.forename: Ruth
authors.initials: R
authors.lastname: Levitz
authors.forename: Rachel
authors.initials: R
authors.lastname: Wattier
authors.forename: Pamela
authors.initials: P
authors.lastname: Phillips
authors.forename: Alexandra
authors.initials: A
authors.lastname: Solomon
authors.forename: Jessica
authors.initials: J
authors.lastname: Lawler
authors.forename: Isaac
authors.initials: I
authors.lastname: Lazar
authors.forename: Carla
authors.initials: C
authors.lastname: Weibel
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Kahn
mesh_terms: D002460:Cell Line; D018946:Chemokine CCL5 / Q000096:biosynthesis*; D002675:Child, Preschool; D004847:Epithelial Cells / Q000276:immunology* / Q000821:virology*; D020869:Gene Expression Profiling; D054884:Host-Pathogen Interactions; D006801:Humans; D007223:Infant; D015850:Interleukin-6 / Q000096:biosynthesis*; D060888:Real-Time Polymerase Chain Reaction; D018357:Respiratory Syncytial Virus Infections / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000276:immunology* / Q000302:isolation & purification / Q000472:pathogenicity*; D014774:Virulence
chemical_list: C483024:CCL5 protein, human; D018946:Chemokine CCL5; C508600:IL6 protein, human; D015850:Interleukin-6