Immunoglobulin light chain repertoire in chronic lymphocytic leukemia

Blood ◽  
2005 ◽  
Vol 106 (10) ◽  
pp. 3575-3583 ◽  
Author(s):  
Kostas Stamatopoulos ◽  
Chrysoula Belessi ◽  
Anastasia Hadzidimitriou ◽  
Tatjana Smilevska ◽  
Evangelia Kalagiakou ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Light Chain ◽  
Lymphocytic Leukemia ◽  
Immunoglobulin Light Chain ◽  
Neoplastic Cells ◽  
Heavy Chains ◽  
Immunoglobulin Kappa ◽  
Complementarity Determining Region ◽  
Immunoglobulin Lambda

AbstractImmunoglobulin kappa (IGK) and immunoglobulin lambda (IGL) light chain repertoire was analyzed in 276 chronic lymphocytic leukemia (CLL) cases and compared with the relevant repertoires from normal, autoreactive, and neoplastic cells. Twenty-one functional IGKV genes were used in IGKV-J rearrangements of 179 kappa-CLL cases; the most frequent genes were IGKV3-20(A27), IGKV1-39/1D-39(O2/O12), IGKV1-5(L12), IGKV4-1(B3), and IGKV2-30(A17); 90 (50.3%) of 179 IGK sequences were mutated (similarity < 98%). Twenty functional IGLV genes were used in IGLV-J rearrangements of 97 lambda-CLL cases; the most frequent genes were IGLV3-21(VL2-14), IGLV2-8(VL1-2), and IGLV2-14(VL1-4); 44 of 97 IGL sequences (45.4%) were mutated. Subsets with “CLL-biased” homologous complementarity-determining region 3 (CDR3) were identified: (1) IGKV2-30-IGKJ2, 7 sequences with homologous kappa CDR3 (KCDR3), 5 of 7 associated with homologous IGHV4-34 heavy chains; (2) IGKV1-39/1D-39-IGKJ1/4, 4 unmutated sequences with homologous KCDR3, 2 of 4 associated with homologous IGHV4-39 heavy chains; (3) IGKV1-5-IGKJ1/3, 4 sequences with homologous KCDR3, 2 of 4 associated with unmutated nonhomologous IGHV4-39 heavy chains; (4) IGLV1-44-IGLJ2/3, 2 sequences with homologous lambda CDR3 (LCDR3), associated with homologous IGHV4-b heavy chains; and (5) IGLV3-21-IGLJ2/3, 9 sequences with homologous LCDR3, 3 of 9 associated with homologous IGHV3-21 heavy chains. The existence of subsets that comprise given IGKV-J/IGLV-J domains associated with IGHV-D-J domains that display homologous CDR3 provides further evidence for the role of antigen in CLL pathogenesis.

Immunoglobulin Light Chain Repertoire in Chronic Lymphocytic Leukemia (CLL): Recognition of Subsets with “CLL-Specific” CDR3 Regions and Associations with Heavy Chains.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 769-769 ◽  
Author(s):  
Kostas Stamatopoulos ◽  
Chrysoula Belessi ◽  
Anastasia Hadzidimitriou ◽  
Evangelia Kalagiakou ◽  
Tatjana Smilevska ◽  
...  
Keyword(s):  
Light Chain ◽  
Lymphocytic Leukemia ◽  
Molecular Processes ◽  
Immunoglobulin Light Chain ◽  
Heavy Chains ◽  
Molecular Features ◽  
Gene Usage ◽  
Region Length ◽  
Unequal Length

Abstract We analyzed immunoglobulin light chain (IgLC) repertoire in a series of 253 typical, unselected CLL cases and compared CLL IgLC sequences to GenBank IgLC sequences from normal, autoreactive and neoplastic cells. The present series included 165 κ- and 88 λ-CLL cases. Twenty-three functional IGKV genes were used in IGKV-J rearrangements in κ-CLL; the most frequent genes were: 3-20/A27 (25 cases), 1-39-1D-39/O2-O12 (19 cases), 4-1/B3 (16 cases), 1-5/L12 (15 cases), 2-30/A17 (13 cases) and 1-8/L9 (10 cases). There were 55/165 unmutated sequences (33%), 44/165 sequences (27%) with 97-99,6% homology to germline and 66/165 sequences (40%) with less than 97% homology. KCDR3 region length ranged from 6–11 (median, 9) aminoacids (aa). N nucleotides (median 3, range 1–12) were detected in 85/165 rearrangements (51.5%). IGKJ3-5 gene usage was observed in 51/165 rearrangements (30%); interestingly, IGKJ3-5 genes were used in 7/8 IGKV3-11 and 4/5 IGKV1-9 rearrangements. Subsets with homologous and “CLL-specific” KCDR3 regions were identified: IGKV2-30, 5 mutated sequences with identical KCDR3 (MQGTYWPYT), 3/5 associated with IGHV4-34 utilizing heavy chains with a similar HCDR3 of 20 aa; IGKV1-39/1D-39, 3 unmutated sequences with identical KCDR3 (QQSYSTTPLT), all associated with IGHV4-39 utilizing heavy chains with a similar HCDR3 of 19 aa; IGKV1-5, 4 unmutated sequences with identical KCDR3 (QQYNSYPWT), 2/4 associated with unmutated IGHV4-39 utilizing heavy chains with a HCDR3 of unequal length. Twenty-six functional IGLV genes were used in IGLV-J rearrangements in λ-CLL; the most frequent genes were: IGLV2-8/1-2 (14 cases), 3-21/2-14 (13 cases), 2-14/1-4 and 1-44/1-16 (7 cases each). There were 24/88 unmutated sequences (27%), 33/88 sequences (37,5%) with 97-99,6% homology to germline and 31/88 sequences (35%) with less than 97% homology. LCDR3 region length ranged from 8-13 aa (median, 11). N nucleotides (median 3, range 1-15) were detected in 42/88 rearrangements (47.7%). The IGLJ1 gene was used in 18/88 rearrangements (20%); all other rearrangements used the IGLJ3*01/*02 genes. Subsets with homologous and “CLL-specific” LCDR3 regions were identified: IGLV1-44, 2 sequences with very similar LCDR3 (AAWDDSLNGP/QV), both associated with IGHV4-b utilizing heavy chains with a similar HCDR3 of 11 aa; IGLV3-21, 7 sequences all with identical LCDR3 (QVWDSGSDHPWV), 3/7 associated with IGHV3-21 utilizing heavy chains with a similar HCDR3 of 9 aa. These results document that IgLC repertoire in CLL is biased by both intrinsic molecular processes as well as selection after LC expression. Genes that have been reported to be overexpressed in the normal and autoimmune disorders were also found to be overrepresented in the CLL repertoire, often with “CLL-specific” molecular features. Finally, the existence of subgroups with homologous CDR3 regions associated with similar heavy chains provides further evidence for the role of antigen selection in CLL pathogenesis.

Analysis of Non-Expressed IGK Locus Rearrangements in Chronic Lymphocytic Leukemia Indicates a Role for Secondary Rearrangements in Shaping the Expressed Immunoglobulin Repertoire.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 972-972 ◽  
Author(s):  
Chrysoula Belessi ◽  
Kostas Stamatopoulos ◽  
Katerina Hatzi ◽  
Anastasia Hadzidimitriou ◽  
Fotini Marantidou ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Light Chain ◽  
Somatic Hypermutation ◽  
Lymphocytic Leukemia ◽  
Receptor Editing ◽  
Rt Pcr ◽  
Stop Codons ◽  
Immunoglobulin Kappa ◽  
Immunoglobulin Repertoire ◽  
Dna Pcr

Abstract Immunoglobulin kappa (IGK) locus rearrangements were analyzed in 164 κ-light chain (LC) expressing chronic lymphocytic leukemia (CLL) cases and 95 λ-LC expressing CLL cases. In κ-CLL, 151 IGKV-J transcripts were successfully amplified by RT-PCR; 147/151 were in frame; four out-of-frame IGKV-J transcripts were heavily mutated and contained one or more stop codons, presumably introduced by somatic hypermutation; in all four cases the corresponding expressed transcript was also mutated. DNA-PCR identified 24/164 κ-CLL cases (15%) with double IGKV-J rearrangements; 9/24 non-expressed IGKV-J rearrangements were in-frame; 3/9 were mutated. The most frequently used IGKV genes in expressed IGKV-J rearrangements were: 3–20, 1-39/1D-39, 1–5 and 4-1; the IGKV4-1 gene was by far the most frequent in non-expressed IGKV-J rearrangements (9/24 cases, 33%). In λ-CLL, a total of 65 IGKV-J rearrangements were amplified in 59/95 cases (62.0%); six cases had two different rearrangements. IGVK4-1 was the most frequently used gene (14/65 sequences, 21.5%), followed by IGKV1-16 (8 cases,12.3%), 1-33/1D-33 and 2-30 (7 cases each, 10.8%). IGKV-J transcripts were detected by RT-PCR in 10/59 λ-CLL cases with IGKV-J rearrangements, of which four were in-frame and six out-of-frame. Seven IGKV-J rearrangements in λ-CLL had less than 100% homology to germline; 3/7 were in-frame; 6/7 patients had mutated IGHV and 5/7 had mutated IGLV genes. In κ-CLL, biased usage of the IGKJ1/IGKJ2 genes was observed both in expressed (69%) and non-expressed rearrangements (78%); in contrast, in λ-CLL, downstream IGKJ (IGKJ3-5) usage was observed in 32/59 sequences (53%). Nonproductive rearrangements involving the kappa deleting element (KDE) that render the IGK locus inactive were also analyzed. In κ-CLL, 22/147 cases (15%) carried IGKV to KDE rearrangements, while 24/147 cases (16.5%) carried IGKJ-C- INTRON (JKI) to KDE rearrangements. In λ-CLL, IGKV-KDE rearrangements were amplified in 55/94 cases (59%); JKI-KDE rearrangements were amplified in 52/94 cases (56%). IGKV1D-43 was the most frequent gene in IGKV-KDE joints in κ-CLL (4/22 cases); in contrast, the most frequent genes in IGKV-KDE joints in λ-CLL were IGKV3-20 and IGKV2-30 (9/55 and 7/55 cases, respectively). In conclusion, the present study confirms IGK locus rearrangements in the vast majority of λ-LC expressing CLL cases. Differential usage of IGKJ genes along with significant IGKV repertoire differences in both IGKV-J and IGKV-KDE rearrangements between κ- and λ-CLL allude to prolonged IGK locus recombination before CLL clonogenic cells became λ-producers. The inactivation of productive and potentially functional IGKV-J joints by secondary rearrangements indicates a role for receptor editing in shaping the expressed IG repertoire in CLL. Finally, the identification of mutated, non-expressed IGKV-J rearrangements both in κ- and λ-CLL might be considered as evidence for secondary rearrangements occurring after the onset of somatic hypermutation, at least in a proportion of cases.

The origin of deletion 22q11 in chronic lymphocytic leukemia is related to the rearrangement of immunoglobulin lambda light chain locus

2013 ◽  
Vol 37 (7) ◽  
pp. 802-808 ◽  
Author(s):  
Marek Mraz ◽  
Katerina Stano Kozubik ◽  
Karla Plevova ◽  
Katerina Musilova ◽  
Boris Tichy ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Light Chain ◽  
Lymphocytic Leukemia ◽  
Lambda Light Chain ◽  
Deletion 22Q11 ◽  
Chain Locus ◽  
Light Chain Locus ◽  
Immunoglobulin Lambda

Evidence for the Significant Role of Immunoglobulin Light Chains in Antigen Recognition and Selection in Chronic Lymphocytic Leukemia

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 780-780
Author(s):  
Anastasia Hadzidimitriou ◽  
Nikos Darzentas ◽  
Fiona Murray ◽  
Tanja Smilevska ◽  
Eleni Arvaniti ◽  
...  
Keyword(s):  
Amino Acid ◽  
Chronic Lymphocytic Leukemia ◽  
Heavy Chain ◽  
Light Chain ◽  
Antigen Recognition ◽  
Lymphocytic Leukemia ◽  
Light Chains ◽  
Light Chain Gene ◽  
Heavy Chains ◽  
Gene Usage

Abstract The chronic lymphocytic leukemia (CLL) immunoglobulin (IG) heavy chain repertoire is known to display biased immunoglobulin variable heavy-chain (IGHV) gene usage, remarkable complementarity determining region 3 (HCDR3) stereotypy as well as distinctive somatic hypermutation (SHM) patterns, at least for subsets of cases. Our aim in the present study was to similarly investigate the IG light chain (LC) genes in terms of mutation frequency and targeting and CDR3 stereotypy to elucidate if the LC may play a significant complementary role in antigen recognition in CLL. We thus examined SHM patterns and secondary rearrangements of the IG LC gene loci in a total of 612 IGKV-J and 279 IGLV-J rearrangements from 725 patients with CLL. Firstly, we observed a highly restricted light chain gene usage in the vast majority of CLL cases with stereotyped HCDR3s. In particular, stereotyped IGHV3-21 CLL cases were characterized by a strikingly biased expression of lambda light chains utilizing the IGLV3-21 gene (36/37 cases of subset#2), whereas all 15 subset #4 cases with stereotyped IGHV4-34 IGs carried an IGKV2-30 rearrangement. In addition, subset-biased light chain CDR3 motifs were identified in groups of sequences utilizing the same IGKV or IGLV gene. For example, all 30 IGKV1-39/1D-39 light chains of subset#1 (using stereotyped IGHV1/5/7 genes) carried notably long KCDR3s (10–11 amino acids) generated by significant N region addition and characterized by the frequent introduction of a junctional proline (26/30 cases). Important differences regarding mutational load were observed in groups of sequences utilizing the same IGKV or IGLV gene and/or belonging to subsets with stereotyped B cell receptors (BCRs). In fact, significant differences were observed with regard to mutational status among groups of sequences utilizing different alleles of certain IGK/LV genes (specifically the IGKV1-5, IGLV1-51 and IGLV3-21 genes). At cohort level, the SHM patterns were typical of a canonical SHM process. A clustering of R mutations in KCDR1 was evident for all IGKV subgroups with the notable exception of the IGKV2 subgroup, which exhibited preferential targeting to the KCDR2, especially in IGKV2-30 rearrangements of cases with stereotyped IGHV4-34/IGKV2-30 BCRs (subset#4). Recurrent amino acid changes at certain positions across the entire IGKV/IGLV sequence were observed at a high frequency (27–67% of cases) in a number of stereotyped subsets, especially those expressing the IGHV3-21/IGLV3-21 BCR (subset #2) and the IGHV4-34/IGKV2-30 BCR (subset #4). Comparison with CLL LC sequences carrying heterogeneous K/LCDR3s or non-CLL LC sequences revealed that these distinct amino acid changes are greatly under-represented in such groups and appear therefore to be “subset-biased”. Finally, a significant proportion of CLL cases (63 cases; 26 kappa- and 37 lambda-expressing) with monotypic LC expression were found to carry multiple potentially functional LC rearrangements. Of note, nineteen of these 63 cases (30%) belonged to subsets with stereotyped BCRs. This finding alludes to the possibility of secondary rearrangements most likely occurring in the context of (auto)antigen-driven receptor editing, particularly in the case of stereotyped subsets. In conclusion, SHM targeting in CLL LCs appears to be just as precise and, most likely, functionally driven as in heavy chains. Secondary LC gene rearrangements and subset-biased mutations in CLL LC genes are strong indications that LCs are crucial in shaping the specificity of leukemic BCRs, in association with defined heavy chains. Therefore, CLL is characterized not only by stereotyped HCDR3 and heavy chains but, rather, by stereotyped BCRs involving both chains, which create distinctive antigen binding grooves.

IG Heavy and Light Chain Variable Genes in Chronic Lymphocytic Leukemia Exhibit Distinct Somatic Mutation Patterns and a Comparable Imprint of Antigen Selection.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1921-1921
Author(s):  
Kostas Stamatopoulos ◽  
Chrysoula Belessi ◽  
Anastasia Hadzidimitriou ◽  
Katerina Hatzi ◽  
Niki Stavroyianni ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Light Chain ◽  
Somatic Mutations ◽  
Somatic Hypermutation ◽  
Multinomial Distribution ◽  
Lymphocytic Leukemia ◽  
Distribution Model ◽  
Mutational Hotspots ◽  
Antigen Selection

Abstract We conducted a parallel analysis of somatic mutations in paired IG heavy (HC) and light chain (LC) chain genes in a series of 253 (165 κ- and 88 λ-LC expressing) chronic lymphocytic leukemia (CLL) cases. IGHV/IGKV/IGLV sequences were 100% homologous to germline in 32/33/27% of cases; had 97–99.7% homology in 50/27/37,5% of cases; less than 97% homology in 18/40/35% of cases. IGHV sequences were generally more mutated than the corresponding IGKV/IGKL sequences (average mutation load for IGHV/IGKV/IGLV genes: 94.4/96.4/96.8); 18/253 cases (7.1%) had more mutated LCs than HCs. The most frequent genes among 100% homologous sequences were IGHV4-39 and IGHV1-69 (10/16 and 9/15 cases, respectively) and IGKV1-39/1D-39 (9/19 cases); among IGLV genes, 3/13 IGLV3-21 sequences were unmutated and 9/13 had >98% homology. The most frequent genes in the “<97%-homology” subgroup were IGHV3-7 and IGHV4-34 (18/19 and 23/27 cases, respectively), IGKV3-20 (12/25 cases) and IGLV2-8 (8/14 cases). Analysis after the multinomial distribution model disclosed significant evidence for positive selection by classical T-dependent antigen from both chains in 43 cases, from HCs only in 53 cases and from LCs only in 16 cases; 27 HC+LC mutated cases (7/27 with <97% homology) had no evidence for antigen selection from either HCs or LCs. Mutations followed a similar pattern, with elevated replacement/silent (R/S) ratios in CDRs (mean, 2.8, 2.6 and 3.7 for IGHV, IGKV and IGLV, respectively) and decreased R/S in FRs (mean, 1.4, 1.5 and 1.6 for IGHV, IGKV and IGLV, respectively). R mutations in FRs usually led to substitution by aminoacids of similar charge and size. The somatic hypermutation process generally spread diversity to the periphery of the CDR1 region. Mutational “hotspots” differed not only between HCs and LCs but among subgroups (eg, IGHV1 vs. IGHV3 vs. IGHV4: thus, S-92 in IMGT-FR3 was mutated in 52/21/5% of IGHV4/3/1 sequences; IGKV1/3 vs. IGKV2 subgroup genes; IGLV1 vs. IGLV3 subgroup genes) or even individual genes, e.g. IGHV1-8, IGHV3-23, IGHV4-34, IGKV2-30, IGLV3-21; in this context, a subgroup of three mutated IGHV4-34 sequences was identified, all in sIgG+ cases, with similar HCDR3 regions and associated with IGKV2-30 κ-LCs with identical KCDR3; furthermore, in the case of IGLV3-21, an already described association with IGHV3-21 HCs was again noted in our series (three cases). Importantly, some IMGT positions were universal “hotspots” (eg. 32 in IMGT-CDR1) or “coldspots” (both invariant C at IMGT positions 23/104 in all sequences; IMGT positions 41, 43, 74, 75 and 102 for all IGHV subgroups; 16, 41, 76, 87, 89 and 98 for all IGKV subgroups; 41, 43, 50, 70, 76, 83, 89, 98, 99 and 102 for all IGLV subgroups). Finally, IGLV sequences showed a lower incidence of CDR1 and FR2/3 mutations compared to IGKV sequences. In conclusion, a complementary imprint of antigen selection in CLL might be witnessed either by IGHV, IGKV or IGLV rearranged sequences. These results provide further evidence for the role of antigen selection in the pathogenesis of at least a proportion of CLL cases.

scholarly journals Preferential linkage of bcl-2 to immunoglobulin light chain gene in chronic lymphocytic leukemia.

1990 ◽  
Vol 171 (2) ◽  
pp. 559-564 ◽  
Author(s):  
M Adachi ◽  
A Tefferi ◽  
P R Greipp ◽  
T J Kipps ◽  
Y Tsujimoto
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Light Chain ◽  
Chromosome Translocation ◽  
Lymphocytic Leukemia ◽  
Chain Gene ◽  
Immunoglobulin Light Chain ◽  
Light Chain Gene ◽  
Flanking Region ◽  
Follicular Lymphomas

Most of human follicular lymphomas possess the t(14;18) chromosome translocation that juxtaposes the IgH gene to the 3' region of bcl-2 in a head-to-tail configuration. Here we show that the rearrangement of the bcl-2 gene occurs in a significant fraction (approximately of 10%) of B cell CLL. In all cases analyzed, breakpoints on chromosome 18 clustered at the 5' flanking region of the bcl-2 gene, and no rearrangements were found at the major or minor breakpoint clustering region (3' region of bcl-2 gene) typical of the t(14;18) chromosome translocation. All of the rearranged bcl-2 genes were juxtaposed with the Ig lambda or K genes in a head-to-head configuration. These results imply that the bcl-2 gene is preferentially linked to the IgL genes in CLL and could function in leukemogenesis.

Evidence for the significant role of immunoglobulin light chains in antigen recognition and selection in chronic lymphocytic leukemia

Blood ◽  
2009 ◽  
Vol 113 (2) ◽  
pp. 403-411 ◽  
Author(s):  
Anastasia Hadzidimitriou ◽  
Nikos Darzentas ◽  
Fiona Murray ◽  
Tanja Smilevska ◽  
Eleni Arvaniti ◽  
...  
Keyword(s):  
Amino Acid ◽  
Chronic Lymphocytic Leukemia ◽  
Somatic Hypermutation ◽  
Significant Proportion ◽  
Lymphocytic Leukemia ◽  
B Cell Receptors ◽  
Immunoglobulin Light Chains ◽  
Heavy Chains ◽  
Gene Usage

Abstract We analyzed somatic hypermutation (SHM) patterns and secondary rearrangements involving the immunoglobulin (IG) light chain (LC) gene loci in 725 patients with chronic lymphocytic leukemia (CLL). Important differences regarding mutational load and targeting were identified in groups of sequences defined by IGKV/IGLV gene usage and/or K/LCDR3 features. Recurrent amino acid (AA) changes in the IGKV/IGLV sequences were observed in subsets of CLL cases with stereotyped B-cell receptors (BCRs), especially those expressing IGHV3-21/IGLV3-21 and IGHV4-34/IGKV2-30 BCRs. Comparison with CLL LC sequences carrying heterogeneous K/LCDR3s or non-CLL LC sequences revealed that distinct amino acid changes appear to be “CLL-biased.” Finally, a significant proportion of CLL cases with monotypic LC expression were found to carry multiple potentially functional LC rearrangements, alluding to active, (auto)antigen-driven receptor editing. In conclusion, SHM targeting in CLL LCs is just as precise and, likely, functionally driven as in heavy chains. Secondary LC gene rearrangements and subset-biased mutations in CLL LC genes are strong indications that LCs are crucial in shaping the specificity of leukemic BCRs, in association with defined heavy chains. Therefore, CLL is characterized not only by stereotyped HCDR3 and heavy chains but, rather, by stereotyped BCRs involving both chains, which generate distinctive antigen-binding grooves.

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