Detection of minimal residual disease in acute lymphoblastic leukemia by in vitro amplification of rearranged T-cell receptor delta chain sequences.
Human T-cell receptor (TCR) delta-chain diversity mainly originates from high junctional variability, since only a limited number of germline elements is available. This extraordinary diversity at the V.J junction, due to the use of two D delta elements and extensive incorporation of N nucleotides, constitutes a specific clonal marker for cell populations exhibiting rearranged TCR delta genes. To this end we amplified in vitro by polymerase chain reaction (PCR) the TCR delta junctional region of five acute lymphoblastic leukemias (ALL), isolated respective DNA fragments, and used them directly as clonospecific probes. The combination of PCR technology and hybridization to clonospecific probes permitted the detection of leukemia DNA at dilution of 1:100,000 in all five cases. Moreover, we were able to investigate one of the ALL patients 11 months after achieving continuous complete remission. Conventional Southern blot analysis failed to detect rearranged TCR genes at this stage. However, residual leukemic cells could readily be detected by PCR technique. We conclude that the strategy proposed here is a very sensitive tool to detect minimal residual disease in a significant proportion of human lymphoid neoplasias.
Hansen-Hagge
T E
TE
Department of Pediatrics II, University of Ulm, FRG.
Yokota
S
S
Bartram
C R
CR
D001483:Base Sequence; D015139:Blotting, Southern; D004273:DNA, Neoplasm / Q000235:genetics; D005784:Gene Amplification; D015329:Gene Rearrangement, T-Lymphocyte; D006801:Humans; D008969:Molecular Sequence Data; D015337:Multicenter Studies as Topic; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis / Q000235:genetics* / Q000276:immunology; D011379:Prognosis; D011446:Prospective Studies; D015183:Restriction Mapping
D004273:DNA, Neoplasm; D015345:Oligonucleotide Probes