Detection of immunoglobulin gene rearrangement in B lymphoid malignancies by polymerase chain reaction gene amplification

Abstract

Immunoglobulin heavy chain gene rearrangement serves as a marker of cell lineage and clonality in B lymphoproliferative disorders. We have used polymerase chain reaction (PCR) gene amplification to detect immunoglobulin gene rearrangements involving VH251, a heavy chain variable region preferentially utilized in B lymphoproliferative disorders. Using synthetic amplimers derived from VH251 and the heavy chain joining region, under conditions of high stringency, a homogeneous VH251-specific fragment of approximately 350 bp could be amplified from leukaemic DNA. Of 53 cases of B lineage acute lymphoblastic leukaemia screened for VH251 rearrangement by PCR, 10 were positive. A background level of VH251 rearrangement could also be amplified from normal peripheral blood and bone marrow DNA, but a VH251 rearranged leukaemic clone representing 0.01% of bone marrow mononuclear cells could be readily detected. The application of PCR to detect immunoglobulin gene rearrangement involving VH251 and potentially other preferentially utilized V regions provides a sensitive method both for tracking malignant B cells and for the study of normal B cell developmental pathways through which B lineage malignancies arise.

Immunoglobulin heavy-chain gene rearrangement in adult acute lymphoblastic leukemia reveals preferential usage of JH-proximal variable gene segments

“…The difference observed between BM or PB and the ALL group reflects a biological rather than a disease-related difference, as we first suggested in a preliminary study. 49 Comparing our results with previous reports, we did not find overuse of V H 5 (previously described as V H 251) in ALL 50 or V H , V H (4-39), V H (4-59), V H , and V H in fetal bone marrow. 28 The V H 6 gene The highly polymorphic human V H region is thought to have arisen prior to racial divergence and has been stable for at least 30 000 years.…”

Molecular monitoring of low grade non-Hodgkin's lymphoma by gene amplification

“…This highlights the need for other targets for amplification in low grade NHL since some 10-15% of tumours do not have a bcl-2 rearrangement involving the MBR or MCR. One target that may prove suitable is the rearranged variable gene of the immunoglobulin heavy chain locus (Deane & Norton, 1990;Yamada et al, 1990).…”

IgM-producing chronic lymphocytic leukemia cells undergo immunoglobulin isotype-switching without acquiring somatic mutations.

“…To determine if a subset of the IgM ϩ CLL cells in these patients undergoes isotype-switching, we used an isotype-specific Ig gene fingerprinting assay to compare the CDR3 lengths of the , ␣, ␥1, ␥2, ␥3, and ␥4 PBL mRNAs (19,22,23). The assay was based on specific amplification of cDNAs belonging to a particular H chain isotype, followed by radioactive labeling of the PCR products by primer-extension with internal isotypespecific oligonucleotides.…”