Korean J Lab Med.  2009 Aug;29(4):269-276. 10.3343/kjlm.2009.29.4.269.

Detection of Putative T cell Clones Using T cell Receptor beta Chain Gene Clonality Assay in Korean Patients with Aplastic Anemia

• Choi HJ
• Shin MG
• Kim HR
• Kim HJ
• Kook H
• Kee SJ
• Kim SH
• Shin JH
• Suh SP
• Ryang DW

• KMID: 1096917
• DOI: http://doi.org/10.3343/kjlm.2009.29.4.269

Abstract

BACKGROUND: We analyzed T cell receptor beta chain (TCRB) gene to investigate the presence of putative T cell clones and its clinicopathologic implications in Korean patients with aplastic anemia (AA).
METHODS
Twenty-nine bone marrow specimens were collected from 20 AA patients, 19 specimens from initial diagnosis and 10 from follow-up. T cell clonality assay was performed using IdentiClone(TM) TCRB Gene Clonality Assay kit (InVivoScirbe Technology, USA) and automatic genetic analyzer. Patients' clinical information and laboratory parameters were also analyzed.
RESULTS
Five patients had definitive underlying factors related with aplastic anemia, such as hepatitis B virus (4 cases) and benzene exposure (1 case). Putative T cell clones were detected in bone marrow specimens of 11 (58%) out of 19 patients at diagnosis. The location of putative T cell clones of TCRB gene (diversity region, Dbeta; joining region, Jbeta; variable region, Vbeta) was distributed in Dbeta2+Jbeta2 (6 cases), Dbeta1+Jbeta1 (3 cases), Vbeta+Jbeta1 (2 cases), and Dbeta1+Jbeta2 (2 cases). Interestingly, among seven patients who underwent stem cell transplantation, five patients with no T cell clones detected at diagnosis developed new T cell clones during the follow-up.
CONCLUSIONS
Putative pathogenetic T cell clones were detected in most of AA patients in the current study. T cell clonality assay would be useful for investigating the pathophysiology of acquired AA.

Keyword

Aplastic anemia; T cell receptor beta chain gene; T cell clone

MeSH Terms

Adolescent
Adult
Aged
Anemia, Aplastic/*diagnosis/genetics/therapy
Bone Marrow Transplantation
Female
Humans
Male
Middle Aged
Reagent Kits, Diagnostic
Receptors, Antigen, T-Cell, alpha-beta/*genetics
Republic of Korea
T-Lymphocytes/*cytology/immunology

Figure

• 

  Fig. 1. Schematic diagram of strategy for T cell clonality assay. A representative rearranged T cell receptor beta gene on chromosome 7 (7q35) shows the approximate placement of the upstream and downstream DNA primers. The numbers of primers and their specificity are listed for master mix tubes A, B and C.

• 

  Fig. 2. Capillary electrophoretogram of amplified T-cell receptor beta chain (TCRB) gene. Positive peaks (presence of clonal cell population) are observed in IVS-0004 positive control DNA at 295 base pairs (bp) (A), and patient No. 13 at 21 days after bone marrow transplantation in TCRB variable and joining region (B). Typical polyclonal peaks are observed in Patient No. 20 at 240-285 bp (C). The definition of positive peaks was at least three times the amplitude of the third largest peak in the polyclonal background within the valid size range.

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