title: A novel approach for the analysis of T-cell reconstitution by using a T-cell receptor beta-based oligonucleotide microarray in hematopoietic stem cell transplantation.
abstract: OBJECTIVE
  Analysis of T-cell population diversity is important to hematopoietic stem cell transplantation (HSCT). The millions of specificities in T-cell receptor (TCR) hypervariable complementarity- determining region 3 (CDR3) precludes detection of all T-cell populations by antibody-based flow cytometry. An alternative method, the TCR CDR3 spectratyping assay, involves multiple polymerase chain reaction (PCR) analyses and is interpreted only qualitatively. In this study, we designed the first TCRbeeta-based oligonucleotide microarray and investigated its specificity, clonality discrimination, sensitivity of detection, and feasibility for monitoring T-cell population diversity in HSCT.
  
  
  METHODS
  The array contains 27 TCR Vbeta probes and 13 Jbeta probes. TCRbeta repertoire diversity was detected with single PCR, microarray hybridization system, and Spotfire analysis software.
  
  
  RESULTS
  TCRO-based microarray provides specific sequence-based information and can distinguish T-cell monoclonal expansion within a polyclonal population. We successfully used this microarray to quantitatively and qualitatively analyze T-cell population diversity in recipients of hematopoietic stem cell transplants.
  
  
  CONCLUSIONS
  This success suggests broad potential applications of the microarray for use in many other areas, including anti-tumor immunity, vaccination, autoimmunity, infectious diseases, and leukemia. By providing a single PCR-based assay to quantify multiple T-cell populations in parallel, this device will allow clinicians and researchers to rapidly perform high-throughput surveys.
authors.affiliation: Division of Bone Marrow Transplantation, St Jude Children's Research Hospital, MaiJ Stop 321, 332 N Lauderdale Street, Memphis, TN 38105-2794, USA. xiaohua.chen@stjude.org
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Chen
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Hale
authors.forename: Geoffrey A M
authors.initials: GA
authors.lastname: Neale
authors.forename: James
authors.initials: J
authors.lastname: Knowles
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Barfield
authors.forename: Yong-Dong
authors.initials: YD
authors.lastname: Wang
authors.forename: Deepak
authors.initials: D
authors.lastname: Kaushal
authors.forename: Deanna C
authors.initials: DC
authors.lastname: Naeve
authors.forename: Deo Kumar
authors.initials: DK
authors.lastname: Srivastava
authors.forename: Xin
authors.initials: X
authors.lastname: Tong
authors.forename: Victoria
authors.initials: V
authors.lastname: Turner
authors.forename: Clayton W
authors.initials: CW
authors.lastname: Naeve
authors.forename: Rupert
authors.initials: R
authors.lastname: Handgretinger
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D002999:Clone Cells; D005434:Flow Cytometry; D006085:Graft Survival / Q000235:genetics / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D012680:Sensitivity and Specificity; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
chemical_list: D016693:Receptors, Antigen, T-Cell, alpha-beta