FISH detection of chromosome 14q32/IgH translocations: evaluation in follicular lymphoma.
A FISH strategy capable of detecting chromosome 14q32 rearrangements involving the IgH locus, including in interphase nuclei, was developed using Ig variable and constant region cosmids from the extremities of the locus in a dual hybridization approach, using signal splitting as evidence of rearrangement. The large size of the locus (1.3 Mb) and the propensity for internal deletion due to physiological VDJ recombination and isotype switching complicate analysis of this locus. We used the Ig10 cosmid, which hybridizes to C epsilon and C alpha2 at the 3' end of the constant region, in order to minimize deletion and/or splitting of the constant region probe. Cos Ig10 and the IgV18 VH probes were compared with a specific IgH-BCL2 FISH dual hybridization approach in follicular lymphoma (FL). Both were capable of detecting the t(14;18) in interphase nuclei, including in cases with no apparent abnormality by classic karyotype analysis, although the sensitivity of the IgH approach was slightly lower. We have also successfully applied these probes to whole cell cytospin preparations, rendering analysis of cryopreserved material possible, although interpretation should be limited to frequent events, particularly following cell manipulation. Analysis of flow cytometric sorted bone marrow fractions from three FL patients by FISH and FICTION showed that the t(14;18) was present in a much lower proportion of CD34 positive than negative cells but that the higher level of background hybridization limits use of these techniques for the reliable quantification of rare events.
Rack
K A
KA
Department of Biological Haematology, Hôpital Necker-Enfants Malades, Paris, France.
Salomon-Nguyen
F
F
Radford-Weiss
I
I
Gil
M O
MO
Schmitt
C
C
Belanger
C
C
Nusbaum
S
S
Vekemans
M
M
Valensi
F
F
Macintyre
E A
EA
D002469:Cell Separation; D002883:Chromosomes, Human, Pair 14; D002887:Chromosomes, Human, Pair 18; D005434:Flow Cytometry; D019254:Genes, bcl-2; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence; D007399:Interphase; D007621:Karyotyping; D008224:Lymphoma, Follicular / Q000235:genetics*; D008677:Metaphase; D014178:Translocation, Genetic
D007143:Immunoglobulin Heavy Chains