Immunoglobulin heavy chain variable region family usage is independent of tumor cell phenotype in human B lineage leukemias

Abstract

During B cell development, immunoglobulin heavy chain (IgH) variable region (VH) genes are rearranged and expressed in a programmed manner and accumulating evidence suggests recurrent utilization of developmentally restricted VH genes in malignant B lymphoid populations. We have used polymerase chain reaction gene amplification in conjunction with a panel of VH family-specific amplimers to directly compare the repertoire of VH region rearrangement in mature, CD5+ B cell chronic lymphocytic leukemia with that in immature, CD5 B lineage acute lymphoblastic leukemia. The results revealed a diverse pattern of VH family utilization common to both disease groups in which VH regions most proximal to the IgH joining locus were preferentially rearranged relative to their family sizes with recurrent utilization of several known developmentally restricted VH genes in close to germ-line configuration. These results indicate that biased VH family usage is independent of tumor cell phenotype in B lineage leukemias. This bias may reflect similar stages or compartments in normal B lymphopoiesis from which diverse types of B cell malignancy may arise. Moreover, since blast cells in acute lymphoblastic leukaemia do not express functional immunoglobulin, we infer that the tumor cell-associated VH family repertoire is determined through antigen-independent mechanisms.

Normal B Cells Express51p1-Encoded Ig Heavy Chains That Are Distinct From Those Expressed by Chronic Lymphocytic Leukemia B Cells

“…2). This mean value is significantly shorter than that previously reported for 51p1-expressing CLL B cells (19.08 Ϯ 3.5: p Ͻ 0.0001, n ϭ 36 unpaired t test) (6,7,14,(17)(18)(19)(20)(21), as well as shorter than these 36 sequences combined with 15 additional 51p1-expressing CLL sequences reported in the literature (18.80 Ϯ 3.2: p Ͻ 0.0001, n ϭ 51 unpaired t test) (6,7,14,(17)(18)(19)(20)(21)(22)(23)(24). The mean CDR3 length of the 39 sequences is also not significantly different from that calculated for the same and similar blood B cell samples using allele-specific PCR (Fig.…”

“…Allelespecific PCR of rearranged 51p1 genes from eight normal blood and tonsillar B cell samples revealed a mean CDR3 length that ranges from 14.6 to 15.5 codons. Both individually and collectively, the mean CDR3 lengths determined using either genomic DNA or cDNA from normal blood or tonsillar B cells were significantly different from the 18.8-codon (Ϯ3.2, n ϭ 51) mean CDR3 length noted for 51p1-encoded Ig expressed by CLL B cells ( p Ͻ 0.0001; see Table I) (6,7,14,(17)(18)(19)(20)(21)(22)(23)(24). However, the calculated mean CDR3 lengths of 51p1-encoded Ig of normal blood B cells were not significantly different from the mean CDR3 lengths of 146 Ig encoded by random V H genes expressed by normal adult B cells (13.9 Ϯ 4.2) (25,26) or the mean CDR3 lengths of CLL B cells that express IgH encoded by V H 1 genes other than 51p1 (7,14,20,22,23).…”

Atiprimod blocks STAT3 phosphorylation and induces apoptosis in multiple myeloma cells

“…We then tested the effects of Atiprimod on the IL-6-responsive myeloma cell line MM-1 (Deane and Norton, 1990). As expected, we found that IL-6 induced STAT3 phosphorylation in a dosedependent manner at concentrations ranging from 0.5 to 2 ng ml À1 .…”

“…To avoid contamination by residual effete myeloma cells, only isolated colonies were microaspirated. The rearranged IgH gene in the DNA from the diagnostic myeloma BM cells and from individual colonies was amplified using a modification of a previously described method (Deane and Norton, 1990). Briefly, cells were placed in 1 ml of QuickExtract DNA Extraction solution 1.0 (Epicentre, Madison, WI, USA) and lysed in accordance with the manufacturer's protocol.…”

Comparative analysis of flow cytometry and polymerase chain reaction for the detection of minimal residual disease in childhood acute lymphoblastic leukemia

“…At diagnosis, we identified IgH gene rearrangements using the consensus V-region primers FR-1C 26 or FR-2B, 27 in conjunction with a universal J H primer. 28 Reaction mixtures of 50 ml contained 1 Â PCR buffer (PE Applied Biosystems, Foster City, CA, USA), 50-200 ng of genomic DNA, 1.5-2.5 mM MgCl 2 , 200 mM dNTPs, 0.25-0.50 mM primers, and 1 U of AmpliTaq Gold (PE Applied Biosystems). PCR products were analyzed on 3% agarose gels and potential rearrangements were analyzed by heteroduplex analysis on 6% acrylamide -1 Â TBE gels.…”