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 V_H251. a heavy chain variable region preferentially utilized in B lymphoproliferative disorders. Using synthetic amplimers derived from V_H251 and the heavy chain joining region, under conditions of high stringency, a homogeneous V_H251-specific fragment of approximately 350 bp could be amplified from leukaemic DNA. Of 53 cases of B lineage acute lymphoblastic leukaemia screened for V_H2 51 rearrangement by PCR, 10 were positive. A background level of V_H251 rearrangement could also be amplified from normal peripheral blood and bone marrow DNA. but a V_H251 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 V_H251 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.