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.
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.
Chen
Xiaohua
X
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
Hale
Gregory A
GA
Neale
Geoffrey A M
GA
Knowles
James
J
Barfield
Raymond C
RC
Wang
Yong-Dong
YD
Kaushal
Deepak
D
Naeve
Deanna C
DC
Srivastava
Deo Kumar
DK
Tong
Xin
X
Turner
Victoria
V
Naeve
Clayton W
CW
Handgretinger
Rupert
R
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*
D016693:Receptors, Antigen, T-Cell, alpha-beta