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KMID : 0350519940470020899
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Journal of Catholic Medical College
1994 Volume.47 No. 2 p.899 ~ p.912
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Identification of Immunogenotype and Detection of Minimal Residual Disease in Childhood Acute Lymphocytic Leukemia by Amplification of Antigen Receptor Genes
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Abstract
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Hybirdoma technology made it possible to produce virtually limitless quantities of highly specific monoclonal antibodies that recognize distinct epitopes of cellular antigens. This advance, combined with molecular probes that identity
antigen-receptor
gene rearrangements, has led to valuable insights into normal lymphocyte differentiation and the cellular origins of lymphoid leukemia. Improved methods of immunogenetic marker evaluation have also increased the precision of diagnosis,
classification
and treatment of the lymphoid malignancies, now permitting the detection of minimal residual disease. The polymerase chain reaction (PCR) was used to study immunogenotype in 29 cases of children with acute lymphocytic leukemia(ALL) and to detect
minimal
residual disease in 4 cases of morphological remission state of B-lineage ALL. We amplified both the complementarity determinig region Ⅲ(CDR Ⅲ) of rearranged immunoglobulin heavy chain(IgH) gene and the junctional site of rearranged T cell
receptor
γ(TCRγ) gene by using double(nested) PCR.
@ES The results were as follows:
@EN 1. PCR with primers corresponding to the consensus sequences of IgH CDR Ⅲ gene in B-lineage ALL patients showed a discrete band of amplified product, 100 to 200 base pairs(bp) in size and PCR with consensus primers of TCRγ gene in T-lineage
ALL
patients showed a discrete band of amplified product, 400 to 500 bp in size.
2. Immunogenotype of 29 cases of childhood ALL were 8 cases of B lineage, 5 cases of T lineage, 12 cases of coross lineage and 4 cases of unknown lineage compared with 21 cases of B lineage, 6 cases of T lineage and 2 cases of cross lineage of
immunophenotype.
3. Immunological markers according to the prognostic factors of ALL such as sex, age, initial leukocyte count and FAB classification showed statistically no significnat differences respectively, but the more incidence of T lineage in
immunogenotype
appeared below 8 years old and L2 of FAB classification.
4. As 10E4 and 10E3 fold dilution of the positive DNA could be amplified in B and T lineage ALL, the sensitivity of PCR technique was 0.01% and 0.1%, respectively.
5. 4. Cases of morphological remission state showed positive band in 2 cases of 4 weeks after induction chemotherapy, but no band in 2 cases of maintained remission for 2.5 years.
These results suggest that immunogenotype can be discriminated by PCR in the majority of children with ALL. PCR amplification of IgH CDR Ⅲgene was highly sensitive method for detecting of the minimal residual disease in B lineage ALL. Also, it
is
necessary that further study must be extended to T lineage lymphoid neoplasms.
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