scholarly journals Immunoglobulin Heavy Chain Gene Rearrangements in Patients with Gaucher Disease

2018 ◽  
Vol 37 (3) ◽  
pp. 307-312
Author(s):  
Predrag Rodić ◽  
Milan Lakočević ◽  
Sonja Pavlović ◽  
Teodora Karan Đurašević ◽  
Tatjana Kostić ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Gaucher Disease ◽  
Pcr Analysis ◽  
Chain Gene ◽  
Gene Rearrangements ◽  
Clonal Rearrangement ◽  
Pcr Products ◽  
Igh Rearrangement ◽  
Monoclonal Proteins

Summary Several studies support the evidence of increased incidence of hematological complications in Gaucher disease including monoclonal and polyclonal gammopathies and blood malignancies, especially multiple myeloma. Serum concentrations of immunoglobulins and PCR analysis of the IGH gene rearrangements were performed. The clonal PCR products were directly sequenced and analyzed with the appropriate database and tools. Serum monoclonal proteins were detected and identified by electrophoresis. Among 27 Gaucher patients, clonal IGH rearrangement was discovered in eight, with 5/8 having also serum monoclonal protein. Elevated immunoglobulins were detected in 9/27 patients. Follow-up data for 17 patients showed that the clonal rearrangement remained the same in four of them, however, in one patient it disappeared after the follow-up period. The remaining 12/17 patients were without previous IGH clonal rearrangement and remained so after the follow-up. Although clonal expansion may occur relatively early in the disease course, at least judging by the IGH gene rearrangements in Gaucher patients, the detected clones may be transient. A careful clinical follow-up in these patients is mandatory, including monitoring for lymphoid neoplasms, especially multiple myeloma.

Mono/Oligoclonal T and NK Cells Are Common in Philadelphia Chromosome Positive (Ph+) Leukemia Patients at Diagnosis and Expand During Successful Tyrosine Kinase Inhibitor Therapy.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 856-856
Author(s):  
Anna Kreutzman ◽  
Vesa Juvonen ◽  
Veli Kairisto ◽  
Marja Ekblom ◽  
Leif Stenke ◽  
...  
Keyword(s):  
T Cell ◽  
Tyrosine Kinase ◽  
Nk Cells ◽  
Research Funding ◽  
Nk Cell ◽  
Gene Rearrangements ◽  
Imatinib Treatment ◽  
Healthy Controls ◽  
Clonal Rearrangement

Abstract Abstract 856 Introduction. Central to current treatment of Ph+ leukemia patients are tyrosine kinase inhibitors (TKIs), which predominantly target the BCR-ABL1 kinase in malignant cells. However, broader-spectrum 2nd generation TKIs, such as bosutinib, dasatinib and nilotinib, also inhibit off-target kinases with important physiological functions. Several in vitro studies have implied that TKIs may have immunosuppressive effects by suppressing activation and proliferation of effector lymphocytes. In contrast, we recently observed immunostimulation during dasatinib therapy in the form of marked expansion of clonal cytotoxic lymphocytes (T- and NK cells) resulting in chronic LGL-type lymphocytosis in peripheral blood (PB). The prevalence, detailed molecular background and clinical implications of clonal lymphocytes during TKI therapy are currently unknown. The aim of this study was to comprehensively analyze clonality and evolution of lymphocyte clones during TKI therapy. Patients and methods. The study population included patients with Ph+ leukemia, both CML (n=28) and ALL patients (n=4) on dasatinib (n=23) and imatinib (n=9) therapies. In addition, samples from 12 healthy controls and diagnostic samples from the nine imatinib treated patients were analyzed. Lymphocyte clonality was determined by analysis of PB mononuclear cells (MNC) for clonal T cell receptor (TCR) γ and δ gene rearrangements by 18 primer pairs covering most known clonal TCR γ and δ rearrangements. Upon positive reaction in heteroduplex analysis, the purified PCR products were sequenced. If clonal rearrangement was observed, allele-spesific PCR primers were designed to allow for quantitative follow-up of lymphocyte clones in each patient. Results. Sequencing-confirmed clonal TCR γ rearrangement was observed only in 1 of 12 healthy controls and no TCR δ gene rearrangements were found in this group. Surprisingly, 7 of 9 (78%) CML patients showed clonal TCR rearrangements at diagnosis. In 3 patients the clonal rearrangement was detected in the TCR δ genes, in 7 patients in the TCR γ genes and 3 patients had rearrangemens both in TCR δ and γ genes. After one year of imatinib treatment the same clones could be detected in 5 of the 7 patients (71%). Although clonal cells were observed, none of the imatinib patients had signs of a concomitant lymphoproliferative disorder and the distribution of lymphocyte subclasses was normal. Next, 23 patients treated with dasatinib were studied, 10 without (LGLneg) and 13 with PB LGL lymphocytosis (LGLpos) including T- or NK-cell expansions. In all LGLpos dasatinib patients (including patients with a CD3neg NK-cell expansion) clonal TCR γ or δ rearrangements were found. In LGLneg dasatinib patients the prevalence of TCR rearrangements was 80%. LGLpos patients had more often clonal rearrangements in TCR δ genes (62%) than LGLneg patients (10%). No differences in clonal rearranged TCR γ genes (77% vs. 80%) were detected. Most patients displayed more than one clonal TCR rearrangement. Quantitative follow-up of LGLpos patients revealed that the expansion of a single predominant lymphocyte clone accounted for LGL lymphocytosis. Intriguingly, quantitative follow-up of lymphocyte clones by PCR showed that the observed clones existed at low levels already before start of dasatinib therapy during imatinib treatment, but no lymphocyte expansions were then seen. Sorting of lymphocytes showed that clonal cells resided in the CD8+ and CD4+ T-cell populations and, strikingly, also among CD16/56+CD3neg NK cells. All dasatinib patients with NK cell expansions (n=3) showed TCR δ rearrangements in their NK cells. Conclusions. Clonal lymphocytes could rarely be found in healthy controls. In contrast, they were frequently present in CML patients at diagnosis and persisted during TKI therapy. In a distinct subgroup of dasatinib treated patients, clonal cells massively expanded during successful therapy. Clonal TCR rearrangements were detected in CD4+, CD8+ and, unexpectedly, also in NK cells. The epitopes and function of clonal, CML-associated lymphocytes are under investigation. Previous studies showed that clonal expansions during dasatinib were associated with excellent, long-lasting therapy responses in advanced leukemia. We therefore hypothesize, that the clonal lymphocytes present at CML diagnosis may be anergic anti-leukemic cells and part of the immune escape mechanisms inherent to leukemogenesis and that dasatinib therapy can reverse this anergy. Disclosures: Ekblom: BMS: Honoraria. Seggewiss:BMS: Honoraria. Porkka:BMS: Honoraria, Research Funding; Novartis: Honoraria, Research Funding. Mustjoki:BMS: Honoraria.

FISH Analysis of IgH Rearrangement by 4 Kinds of Probes Involving IgH Genes in Multiple Myeloma: Improved Sensitivity for the Detection by Use of IGH Break-Apart Probe.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4134-4134
Author(s):  
Hye Yoon Chung ◽  
Miyoung Kim ◽  
Cha Ja See ◽  
Hyun Jung Min ◽  
Sung-Soo Yoon ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Gene Rearrangement ◽  
Chain Gene ◽  
Fish Analysis ◽  
Dual Color ◽  
Igh Genes ◽  
Detectable Rate ◽  
Igh Gene Rearrangement ◽  
Chain Gene Rearrangement ◽  
Igh Rearrangement

Abstract Background: IgH (immunoglobulin heavy chain) gene rearrangement is known to be the most frequent chromosome change in multiple myeloma. The detection of this genetic change is conveniently done by using fluorescence in situ hybridization (FISH) method recently. The aim of this study is to determine the utility of most commonly used probes, IGH/CCND1 dual color, dual fusion probe (Downers Grove, IL, USA), IGH/BCL2 dual color, dual fusion probe (Downers Grove, IL, USA), IGH/FGFR3 dual color, dual fusion probe (Downers Grove, IL, USA) and IGH dual color break apart rearrangement probe from Vysis Products (Downers Grove, IL, USA). Methods: We applied four different probes of IgH FISH on 202 Korean patients with multiple myeloma for comparing the utility of four probes. Results: 84 of 202 patients (41.6%) had the IgH gene rearrangement. 44 of 84 patients (52.4%) showed positive to all four probes, but 40 of 84 patients (47.6%) showed discrepancy. IGH dual color break apart rearrangement probe showed highly detectable rate (41.6%) compare to IGH/CCND1 (31.2%), IGH/BCL2 (27.7%) and IGH/FGFR3 (24.3%). Among patients who showed discrepancy between four probes, 20 of 40 patients (50.0%) were only positive to IGH dual color break apart rearrangement probe. Conclusions: The IGH break-apart probe was qualitatively and quantitatively better than three other probes at initial diagnosis and during follow-up of myeloma. In conclusion, it would be most efficient and advisable to use the IGH break-apart probe at first, and then to identify the translocation partner in case of positive rearrangement of IgH.

Cell-Free DNA for Minimal Residual Disease Monitoring in Multiple Myeloma Patients

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3423-3423 ◽  
Author(s):  
Lenka Kubiczkova-Besse ◽  
Daniela Drandi ◽  
Lenka Sedlarikova ◽  
Stefania Oliva ◽  
Manuela Gambella ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Residual Disease ◽  
Patient Specific ◽  
Cell Free Dna ◽  
Qrt Pcr ◽  
Free Dna ◽  
Diagnostic Samples ◽  
Igh Rearrangement ◽  
Minimal Residual

Abstract Background Circulating nucleic acids, such as cell-free DNA (cf-DNA), are becoming a promising minimally-invasive diagnostic tool for cancer detection. Recent studies demonstrated that tumor-derived cf-DNA can be used to monitor tumor burden and response to treatment in patients (pts) with solid tumors as well as hematological malignancies (Dawson et al, 2013, Armand et al, 2013). In this study we investigated the clinical utility of cf-DNA in the monitoring of minimal residual disease (MRD) of pts with multiple myeloma (MM) carrying the tumor specific immunoglobulin (IGH) rearrangement. Methods Cf-DNA was extracted from 1 ml of serum sample from 13 MM patients enrolled in Italian CRD/MEL-200 and EMN-02 protocols. The total amount of cf-DNA was estimated by fluorometric measurement (median 560 ng, range 15-5158 ng) and the length of fragments was evaluated by high sensitivity dsDNA chips (Agilent). Patient specific clonal IGH rearrangement was identified at the time of diagnosis from bone marrow (BM) genomic DNA (gDNA) as previously reported (Ladetto et al, 2000). For each patient, MRD in BM and peripheral blood (PB) was estimated by real time quantitative PCR (qPCR) using ASO-specific primers and the quantification was based on serial 10-fold dilution standard curves from plasmid carrying the patient specific IGH rearrangement. The amount of IGH rearrangement in cf-DNA (cf-IGH) was estimated by qPCR and droplet digital PCR (ddPCR) (Bio-Rad) on diagnostic and follow up samples and was expressed as the amount of copies per 1 µg of total cf-DNA. qPCR and ddPCR results were interpreted according to the Euro-MRD guidelines (van der Velden et al, 2003). Results Overall, 54 cf-DNA samples from MM serum (13 diagnostic, 41 follow-up samples) were analyzed for the presence of patient specific IGH rearrangement. The most abundant fraction of cf-DNA was 180-220bp, than 350-400bp and 700-10000bp (in 100%, 85% and 68% of samples respectively), whereas longer fragments more often appeared in follow-up samples. By qPCR, cf-IGH at diagnosis were observed in 11/13 diagnostic samples. Only 3/13 pts were quantifiable (116, 85, 187 copies/1 µg of cfDNA) and 8/13 pts were positive but not quantifiable (PNQ) cf-IGH. By ddPCR, levels of cf-IGH at diagnosis were observed in 9/13 pts. 6/13 pts were quantifiable (246, 195, 96, 88, 184, 25 copies/1µg of cfDNA), and only 3/13 pts were PNQ. In follow-up samples, levels of cf-IGH were undetectable by qRT-PCR; however in 5 samples they were PNQ by ddPCR. Interestingly, in one available relapse sample, cf-IGH reappeared again to quantifiable level (61 copies by qRT-PCR and 190 copies by ddPCR). The levels of cf-IGH are quantifiable in samples with higher amount of tumor specific IGH rearrangements in BM or PB; however, no association was observed between cf-IGH level at diagnosis and disease burden estimated by the PCs infiltration in BM or the monoclonal immunoglobulin concentration in blood/urine. Conclusions These data show the potential utility of cf-IGH monitoring in MM pts. Although by qPCR, cf-IGH were detected in 11/13 pts, they were quantifiable only in 3/13 pts and ddPCR was more precise as it was able to quantify cf-IGH in 6/13 pts. Since cf-IGH copies were quantifiable only in diagnostic samples and in 1 available sample at the relapse, we conclude that higher amounts of serum are necessary to overcome the limitation of assay sensitivity. Potential advantages and predictive value, for monitoring tumor marker in a non-invasive manner, need to be further validated on larger cohort of samples using increased amount of cf-DNA. Work was supported by IGA grants NT12130, NT14575. This work is funded by a Black Swan Research Initiative grant by the International Myeloma Foundation "Dynamics of microRNA and cell-free DNA profiles during multiple myeloma progression“. Disclosures Boccadoro: Celgene: Honoraria; Janssen: Honoraria; Onyx: Honoraria. Palumbo:Amgen: Consultancy, Honoraria; Bristol-Myers Squibb: Consultancy, Honoraria; Array BioPharma: Honoraria; Genmab A/S: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Janssen-Cilag: Consultancy, Honoraria; Millennium Pharmaceuticals, Inc.: Consultancy, Honoraria; Onyx Pharmaceuticals: Consultancy, Honoraria; Sanofi Aventis: Honoraria.

scholarly journals PCR Analysis of Immunoglobulin Heavy Chain (IgH) and TcR-γ Chain Gene Rearrangements in the Diagnosis of Lymphoproliferative Disorders: Results of a Study of 525 Cases

2000 ◽  
Vol 13 (12) ◽  
pp. 1269-1279 ◽  
Author(s):  
Catherine Thériault ◽  
Sandrine Galoin ◽  
Severine Valmary ◽  
Janick Selves ◽  
Laurence Lamant ◽  
...  
Keyword(s):  
Heavy Chain ◽  
Immunoglobulin Heavy Chain ◽  
Lymphoproliferative Disorders ◽  
Pcr Analysis ◽  
Chain Gene ◽  
Gene Rearrangements

scholarly journals Advancement of multiple myeloma from diagnosis through plateau phase to progression does not involve a new B-cell clone: evidence from the Ig heavy chain gene

Blood ◽  
1993 ◽  
Vol 82 (1) ◽  
pp. 202-206 ◽  
Author(s):  
QM Ralph ◽  
MJ Brisco ◽  
DE Joshua ◽  
R Brown ◽  
J Gibson ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
B Cell ◽  
Heavy Chain ◽  
Cell Clone ◽  
Neoplastic Cell ◽  
Chain Gene ◽  
Plateau Phase ◽  
Igh Genes ◽  
Igh Rearrangement ◽  
Ig Heavy Chain

The Ig heavy chain (IgH) gene was used as a marker to investigate clonal succession and the origin of the neoplastic cell in multiple myeloma. The polymerase chain reaction (PCR) was used to amplify a section of the rearranged IgH gene at diagnosis and at progression in 21 patients who had exhibited a plateau phase. A monoclonal PCR product was seen for 16 of the patients and the product present at progression was of the same molecular weight as that at diagnosis. This finding suggests that the IgH rearrangement present at diagnosis and progression was the same. This was confirmed by sequencing the IgH gene in 10 patients. The IgH genes were found to be hypermutated at diagnosis, but no further hypermutation occurred during the course of the disease. The results provide evidence that the neoplastic cell in myeloma may originate as a memory B cell, plasmablast, or plasma cell, and suggest that progression beyond the plateau phase is not caused by clonal succession.

Long-Term Follow-Up of Patients with T-Large Granular Lymphocyte Leukemia (T-LGL); Experience in a Single Institution.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5026-5026
Author(s):  
Ahmed Aribi ◽  
Michael Keating ◽  
Susan O’Brien ◽  
Alessandra Ferrajoli ◽  
Stefan Faderl ◽  
...  
Keyword(s):  
Respiratory Infections ◽  
Cell Receptor ◽  
Treatment Modalities ◽  
Cancer Center ◽  
Pcr Analysis ◽  
Chain Gene ◽  
Gamma Chain ◽  
Lymphoid Leukemia ◽  
Minor Response

Abstract T-LGL is a clonal indolent lymphoid leukemia characterized by accumulation of large granular T-cells and cytopenias. We have identified 21 patients with T-LGL who were seen at the MD Anderson Cancer Center between 1997 and 2005. The median age at diagnosis was 53.5 years (range 34 to 72) without a male or female predilection. Fourteen patients (67%) were asymptomatic at diagnosis. Five patients (24%) presented with fatigue secondary to anemia; 1 (5%) had symptomatic splenomegaly and 1 presented with a diverticular abscess. Eighty percent of the patients presented with anemia with median hemoglobin of 9.9g/dl (range 7.4 to12.9 g/dl), 47% with neutropenia with a median absolute neutrophil count of 975/ul (range 110 to 5600/ul), and 38% had thrombocytopenia with a median platelets count of 61.5 x 109/l (range18 to 135 x 109/l). Six patients (29%) had splenomegaly, 3 (14.2%) had hepatomegaly and 1 had retroperitoneal lymph node enlargement on CT. During the course of follow-up (median 19 months, range 2–78 months), 11 patient (52%) developed constitutional symptoms. Six patients had recurrent infections including upper respiratory infections, pneumonia, and folliculitis unrelated to initiation of treatment. Associated autoimmune conditions included rheumatoid arthritis in 3 patients (14%) and hemolytic anemia in one patient. Only two patients had cytogenetic abnormalities, both including abnormalities involving chromosome 6. Monoclonal T cell receptor gamma chain gene rearrangements were detected by PCR analysis in 17 of 18 evaluable patients. Immunophenotypic studies revealed 16 patients to be CD4−/CD8+, 3 CD4−/CD8−, 1 CD4+/CD8+, and 1 CD4+/CD8−. CD56 was positive in 1 patient and none expressed CD26. Five patients (24%) have required no therapy, and remain asymptomatic with a median follow up of 67 months (range 5 to 78). Three patients received erythropoietin or G-CSF with some improvement. Nine patients (43%) were treated with cyclosporine, 2 achieving a response, 2 with stable disease, 3 with progression, and 2 were lost follow up. One of the 3 patients with disease progression had splenectomy and 1 was treated with infliximab; both responded. Combination of pentostatin and alemtuzumab was used in 2 patients with no improvement in the cytopenias; therapy was discontinued in both due to worsening of cytopenias and infections. Other treatment modalities included alemtuzumab alone in 1 patient with no improvement, fludarabine in 1 with minor response, combination of fludarabine and cyclophosphamide in 1 with minor response, methotrexate in 2 with no response and splenectomy in 3 with response and no requirements for further intervention. We conclude that T-LGL generally has an indolent course with no need for active therapy; patients with progressive cytopenias or refractory disease may benefit from splenectomy. New treatment modalities are needed.

Aggregation of Serum Free Light Chains (FLC) Causes Overestimation of FLC Nephelometric Results as Compared to Serum Protein Electrophoresis (SPE) While Preserving Clinical Usefulness.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4767-4767
Author(s):  
Jean-Pierre Émond ◽  
Stephen Harding ◽  
Bernard Lemieux
Keyword(s):  
Molecular Weight ◽  
Multiple Myeloma ◽  
Gel Filtration ◽  
Added Value ◽  
High Dose ◽  
Serum Free ◽  
M Spike ◽  
Pass Through ◽  
Monoclonal Proteins

Abstract Multiple myeloma is a disease characterised by the overproduction of monoclonal proteins. Whilst concentration of FLC correlates poorly with tumour burden at diagnosis, changes during treatment are considered to be an accurate and sensitive measure of response. Aggregation of FLC presents a problem for traditional means of FLC estimation, large aggregates will not pass through the kidney making urine unreliable. Objectives : To report follow-up of 2 patients with discrepancy between SPE M-spike and serum FLC, one with serum free lambda, the other with serum free kappa. To analyze and confirm presence of multimer FLC aggregates in these 2 patients. Methods : Routine SPE and immunofixation were performed by capillary electrophoresis (CZE). Serum FLC was measured by nephelometry. FLC polymerisation studies were undertaken by SDS-PAGE, Western blot and Sephadex S-200 gel filtration chromatography. Results: First patient (free lambda) is a 80 y woman with IgA lambda multiple myeloma. Upon treatment, IgA lambda peak fell from 18 g/L down to < 0,4 g/L with a serum free lambda remaining at 10 700 g/L. Monoclonal free lambda peak at CZE was estimated to 1,5 g/L, a 7x discrepancy. High dose Decadron therapy was unsuccessful, serum free lambda remaining about 2500 to 4000 g/L for the next 6 months. Polymerisation studies revealed multiple aggregates of FLC with molecular weight (MW) ranging from 24 000 Da to > 200 000 Da. Second patient (free kappa) is a 70 y old man followed since 1985 for an IgG kappa indolent multiple myeloma. Treatment with melphalan in 1998 caused disappearance of M-spike at SPE and persistant hypogammaglobulinemia. Introduction of FLC testing in August 2006 revealed an unexpected serum free kappa of 2740 g/L which remained stable in the following months. This serum free kappa is not visible by CZE. Polymerisation studies revealed multiple aggregates of FLC with MW ranging from 24 000 Da to > 200 000 Da. Conclusion : Serum FLC measurement is a new biological parameter for investigation and follow-up of monoclonal gammopathies with added value as showed here for 2 myeloma treated patients. One should be aware of well known interferences in FLC measurement such as renal dysfunction and polyclonal hyper-gammaglobulinemia. Here we demonstrated a less well documented confounding factor creating an unexpected discrepancy between this nephelometric technique and traditional CZE/SPE. We showed highly overestimated FLC by nephelometry. We revealed a profile of multiple complexes in 2 cases of FLC (one free lambda and one free kappa) of molecular weight up to > 200 000 Da. These multimers may well impact kidney function and interfere with standard urine investigation. It appears clear to us that an interpretation from an experienced professional should accompany any FLC measurement.

scholarly journals Proinflammatory Cytokines Profile, a Link Between Multiple Myeloma and Gaucher Disease

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4952-4952
Author(s):  
Javier Gervas ◽  
Marcio M Andrade ◽  
Pilar Giraldo
Keyword(s):  
Multiple Myeloma ◽  
Proinflammatory Cytokines ◽  
Gaucher Disease ◽  
Conflicts Of Interest ◽  
Analytical Data ◽  
Critical Factor ◽  
Cytokine Analysis ◽  
Significant Difference ◽  
Skeletal Complications

Abstract Gaucher disease (GD) is characterized by a latent chronic inflammatory macrophage activation expressed by an imbalance of pro-inflammatory cytokines, hyperferritinemia, hypergammaglobulinemia, altered calcium homeostasis and metabolic syndrome. The incidence of Monoclonal Gammopathy of Uncertain Significance (MGUS) and multiple myeloma (MM) is increased in GD1 patients (3.5-12.5). The hypothesis is that deregulation of immune system resulting from glucocerebroside accumulation may influence the development of hematological malignancies. However, the pathological mechanisms that influence in MM development in GD1 patients are unknown. Cytokines produced secondary to glucocerebroside accumulation, may be a critical factor to explain the pathogenesis of MM in GD. This cytokine environment could trigger lymphocyte expansion and ultimately transformation. Objective: To analyze and compare the proinflammatory cytokines profile among MGUS, MM and GD1patients. Methods: Between February-May 2014 we have analyzed a panel of eight cytokines (IL4, IL6, IL7, IL10, IL13, MIP1α, MIP1β and TNF) by Luminex®100 platform and Millipore cytokine kits. The analysis was performed in plasma samples stored at diagnosis in Aragon Biobank. We have studied four cohorts: 71 healthy controls, 38 GD1 patients (36.8% women, mean age 39.2 years, range: 17-79), 36 MGUS (58.3% women, mean age: 75 years, range 53-89), 11 IgGk; 6 IgGL; 5 IgAk; 8 IgAL; 3 IgMk; 3 doble IgG+IgA and 49 MM (42% women, mean age 68 years, range 39-89), 20 IgGk; 5 IgGL; 8 IgAk; 5 IgAL, 8 ligth chain disease and 3 doble IgG+IgA. Concerning ISS classification 44.0% MM patients had a score 1, 24.3% score 2 and 31.7% score 3. All patients with MGUS or MM had been diagnosed consecutively during 2009 and followed until now. Clinical and analytical data of MGUS and MM were obtained from the records of the Hematology Department; and the data of GD1 patients from the Spanish Registry (FEETEG). Data were analyzed using non-parametric tests (Mann-Whitney-U and Kruskall-Wallis). Results: After 5 years in follow-up 21 MM patients had died (42.8%) and 8 in the MGUS group (22.2%), 4 MGUS developed a MM (11.1%); and 8 another neoplasia (22.2%); in MM patients 7 developed a second tumor (14.2%). Between GD1 patients one developed a MM and two a solid tumor (7.9%). Concerning the cytokine analysis, a significant difference (p<0.05) was observed in IL4; MIP-1α; MIP-1β and TNFα values between controls; MGUS; MM and GD1 patients; and also IL13 in MM and GD1. MM and GD1 not showed differences in the median values of IL4 and IL13. These groups showed significant differences in MIP-1α, MIP-1β and TNFα values. In MM there are significant differences in IL4 (p=0.012) and MIP-1α (p=0.022) between ISS 1 and ISS 3. In addition there are significant differences in MIP-1α between IgGk vs IgAL and MGUS patients that will be transformed to MM (p=0.018).(See table 1)There are not significant differences in cytokines profile between patients who developed a second neoplasia. Conclusions: These results support that GD1 and MM patients share similar proinflammatory profiles. The chronic cytokines increase in GD patients would contribute to the development of malignancies in these patients. GD1 and MM share skeletal complications, MIP-1α as osteoclast-activating factor could be a good biomarker during follow-up of GD1 or MGUS to detect patients in risk to develop bone complications. MIP-1α is described as significantly elevated in osteopenic MGUS patients(1). Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Histopathologic and Molecular Features of Persistent Polyclonal B-Cell Lymphocytosis Challenging the Benign Nature of the Disorder.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4612-4612
Author(s):  
Ilaria Del Giudice ◽  
Stefano A. Pileri ◽  
Maura Rossi ◽  
Elena Sabattini ◽  
Cristina Campidelli ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Pcr Analysis ◽  
White Pulp ◽  
Splenic Marginal Zone Lymphoma ◽  
Molecular Features ◽  
Lymphoid Infiltration ◽  
Igh Rearrangement

Abstract Persistent polyclonal B-cell lymphocytosis (PPBL) is an unusual expansion of B lymphocytes, whose immunophenotype resembles that of memory B cells. The physiopathology of PPBL is unknown and whether this syndrome represents a pre-malignant or benign disease is unclear. Aiming at further defining the histological features of BM involvement and at challenging the polyclonal nature of PPBL, we reviewed 6 cases of PPBL who underwent BM biopsy. There were 3 males and 3 females, with a median age at diagnosis of 35 years (range 17–46). The median lymphocyte count was 5.3 × 109/L (5.1–8.3). One patient had been splenectomized 10 years earlier and 2 were splenomegalic. During the follow-up (median 20 months, range 5–138), 1 showed progressive spleen enlargement and 2 additional patients developed splenomegaly. Screening for hepatitis B, C and HIV was negative. PB morphology showed 2 to 32% of cleaved lymphocytes. Immunophenotype documented 21 to 70% B lymphocytes in both PB and BM aspirate, all positive for CD19, CD22, CD79b, FMC7, CD18 and CD11a, negative for CD5, CD23, CD10, with no Ig light chain restriction. CD25 was expressed in 4 cases. PCR analysis of the IgH gene configuration of PB and BM cells showed a polyclonal picture in all cases. On Giemsa-stained sections of BM trephines, BM was normocellular in 5 cases and slightly hypocellular in 1. A mild lymphoid infiltrate was visible, mainly interstitial with occasional small lymphoid aggregates. An intrasinusoidal pattern was seen in 4/6 cases. Immunohistochemistry confirmed the interstitial infiltration by B cells (CD79a+, CD20+, IgM+, IgD+, DBA44−/+, CD5−, AnnexinD1−) in all cases and highlighted the intrasinusoidal pattern in 4 in which a splenic marginal zone lymphoma (MZL)-like morphology was described. By IgH gene PCR on BM tissue, a clonal rearrangement was detected in 2/4 cases with intrasinusoidal infiltration; in 2 cases DNA quality was poor. The 4 cases showed splenomegaly at diagnosis or during the follow-up. Splenectomy was performed in 1 patient due to progressive enlargement of the spleen over 9 years. White pulp was normal and red pulp was expanded by an infiltration of small lymphocytes with an intrasinusoidal distribution. Splenic B lymphocytes were CD79a+, CD20+, IgM+, IgD+, DBA44−, as in the BM biopsy, with low proliferation activity and proven clonality by IgH-PCR. In the remaining 2 cases, in which no intrasinusoidal lymphoid pattern was evident, clonal IgH rearrangement was absent and no splenomegaly has developed. We confirm the interstitial lymphoid infiltration of BM trephine in all cases of PPBL, with an evident intrasinusoidal component in some. The detection of IgH rearrangements on lymphocytes infiltrating the BM from bioptic samples and the absence of clonality in PB lymphocytes and in the BM aspirate, possibly due to the intrasinusoidal distribution, have not been described. These findings suggest that at least some of the cases described as PPBL are pre-malignant disorders close to splenic MZL, justifying the occurrence of progressive splenomegaly over time. Prolonged follow-up of patients with PPBL and an accurate biologic work-up of the BM, are therefore warranted.

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