title: Detection of T-cell receptor gene rearrangement in children with Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis using the BIOMED-2 multiplex polymerase chain reaction combined with GeneScan analysis.
abstract: BACKGROUND
  Hemophagocytic lymphohistiocytosis (HLH) is a serious immune disorder. Epstein-Barr virus (EBV) is the most common trigger of secondary HLH. T-cell receptor (TCR) gene rearrangement is detectable in half of patients with EBV-associated HLH using Southern blotting and conventional polymerase chain reaction (PCR) analyses. However, its clinical significance is unclear. The impact of TCR gene clonality on the outcome of patients with EBV-HLH should be evaluated using more sensitive methods. In this study, we used BIOMED-2 multiplex PCR and GeneScan analysis to evaluate TCR gene rearrangement in childhood EBV-HLH.
  
  
  METHODS
  Six children with EBV-HLH were enrolled in this study. EBV load in blood was quantified by real-time PCR. TCR gene rearrangement was analyzed by BIOMED-2 multiplex PCR.
  
  
  RESULTS
  All 6 patients showed monoclonal peaks in TCRβ and/or TCRγ at diagnosis. Serial monitoring of one patient disclosed a change in the rearrangement pattern of the TCR genes in response to immunochemotherapy.
  
  
  CONCLUSIONS
  These findings suggest that BIOMED-2 multiplex PCR, a highly sensitive method for detecting T-cell clonality, is useful for predicting the therapeutic response in childhood EBV-HLH.
authors.affiliation: Department of Laboratory Medicine, Shinshu University Hospital, Asahi, Matsumoto, Japan.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsuda
authors.forename: Yozo
authors.initials: Y
authors.lastname: Nakazawa
authors.forename: Ryu
authors.initials: R
authors.lastname: Yanagisawa
authors.forename: Takayuki
authors.initials: T
authors.lastname: Honda
authors.forename: Eiichi
authors.initials: E
authors.lastname: Ishii
authors.forename: Kenichi
authors.initials: K
authors.lastname: Koike
mesh_terms: D000293:Adolescent; D001483:Base Sequence; D015139:Blotting, Southern; D002648:Child; D017931:DNA Primers; D005260:Female; D015329:Gene Rearrangement, T-Lymphocyte / Q000235:genetics*; D004854:Herpesvirus 4, Human / Q000472:pathogenicity*; D006801:Humans; D007223:Infant; D051359:Lymphohistiocytosis, Hemophagocytic / Q000235:genetics / Q000276:immunology / Q000821:virology*; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers