Southern blot detection of immunoglobulin lambda light chain gene rearrangements for clonality studies.
Southern blot analysis of immunoglobulin (Ig) genes has proven to be important for detection of clonal rearrangements in patients with lymphoproliferative diseases. To improve the detection of clonal Ig lambda (Ig lambda) gene rearrangements, we carefully determined the precise restriction map of the J-C lambda gene region, developed a suitable C lambda probe (IGLC3), and evaluated relevant restriction enzymes in combination with the IGLC3 probe. For the latter purpose, we selected 75 B cell malignancies with proven expression of Ig lambda protein chains in order to be sure that each malignancy contained at least one clonally rearranged IG lambda allele. Our extensive Southern blot analyses with the IGLC3 probe in EcoRI and/or HindIII digests revealed that combined EcoRI/HindIII digestion detected Ig lambda gene rearrangements in 95% of the 75 patients and 94% of the 98 rearranged alleles. In contrast, HindIII and EcoRI single digests allowed detection of rearrangements in only 78% and 83% of the patients, and 67% and 79% of rearranged alleles, respectively. We conclude that the use of the IGLC3 probe in combined EcoRI/HindIII digests is superior to EcoRI and HindIII single digests. This probe/enzyme combination is informative for clonality studies in approximately 95% of patients.
Tümkaya
T
T
Department of Immunology, Erasmus University Rotterdam, University Hospital Dijkzigt, The Netherlands.
Comans-Bitter
W M
WM
Verhoeven
M A
MA
van Dongen
J J
JJ
D000483:Alleles; D001483:Base Sequence; D015139:Blotting, Southern / Q000379:methods*; D015322:Gene Rearrangement, B-Lymphocyte; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007146:Immunoglobulin lambda-Chains / Q000235:genetics*; D016393:Lymphoma, B-Cell / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data
D007147:Immunoglobulin Light Chains; D007146:Immunoglobulin lambda-Chains