[T cell receptor gene recombination in human T-cell leukemia cell line jurkat].
OBJECTIVE
Recently, recombination activating gene (RAG)-mediated transposition has been found to contribute to chromosomal translocation and may be related to the occurrence of lymphoid malignancy; however, the underlying mechanism is unclear. Our previous study showed a co-expression of RAG1 and RAG2 in a human T-cell leukemia cell line Jurkat, which represents a mature stage of T cell development, and that the mRNA levels could be regulated by T cell activators. This study was to determine RAGs-mediated T cell receptor(TCR) gene recombination in Jurkat cells, and thus to provide a new thoughtway for studying the correlations of TCR gene recombination to lymphoid malignancy.


METHODS
TCR Dbeta-Jbeta signal joint T-cell receptor excision DNA circles (sjTRECs) were determined by nested and semi-nested polymerase chain reaction (PCR). Double-strand DNA breaks at recombination signal sequences (RSSs) in the TCR beta chain locus were detected by ligation-mediated polymerase chain reaction (LM-PCR). TdT and Ku70/Ku80 were detected by reverse transcription-polymerase chain reaction (RT-PCR).


RESULTS
Characteristics of junctional diversity of Dbeta2-Jbeta2 sjTRECs and ds RSS breaks associated with Dbeta2 5'-site and Dbeta2 3'-site were detected in DNA of Jurkat cells. TdT and Ku70/Ku80 were also detected.


CONCLUSIONS
There is an ongoing TCR gene recombination in Jurkat cells. Jurkat may be used as an ideal cell line model for studying the correlation of TCR gene recombination to lymphoid malignancy.
Zou
Hong-Yun
HY
Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou, Guangdong, 510515, P. R. China.
Ma
Li
L
Luo
Wei
W
Wang
Xiao-Ning
XN
D034961:Antigens, Nuclear / Q000378:metabolism; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D053903:DNA Breaks, Double-Stranded; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D019672:Genes, T-Cell Receptor; D006801:Humans; D019169:Jurkat Cells; D000072200:Ku Autoantigen; D015458:Leukemia, T-Cell / Q000235:genetics*; D008969:Molecular Sequence Data; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D011995:Recombination, Genetic
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D004247:DNA; D004253:DNA Nucleotidylexotransferase; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen