B-cell Chronic Lymphoproliferative Disorders Research Articles

French National Retrospective Cohort of Hairy-Cell Leukemias: Risk of Second Malignancies after 10 Years of Follow-up

Introduction: Hairy-cell leukemia (HCL) is a chronic B-cell lymphoproliferative disorder with a favorable outcome thanks to treatment with purine analogues (PNA) like cladribine and pentostatin. Here, we updated the French national retrospective cohort of HCL after 10 years of follow-up, in order to evaluate the risk of second cancers in these patients. Methods: Data were collected up to June 2018 through a questionnaire sent to the members of the Société Française d'Hématologie, and centralized in the cohort database. We described the second malignancies observed during the follow-up, distinguishing second 'solid' cancers from second hematological malignancies. Then, using a Fine and Gray model, we performed a multivariate analysis in order to identify second cancer risk factors. Finally, to evaluate the excess of cancers in our cohort in comparison with the French general population, we calculated the standardized incidence ratio (SIR). Results: 279 patients (pts) from 19 centers were included in our retrospective cohort. The median age was 59 years old (range 29-88). 21% had an infectious disease at diagnosis, 23% had a familial history of cancer and 11% a personal history of cancer before HCL diagnosis. The median number of lines of treatments was 1 (0-7). PNA (cladribine or pentostatin) were the first therapeutic choice in frontline (75% of pts) and at relapse (69%). With a median follow-up of 127 months (2-413), the median overall survival for the overall study population was 328 months (95% CI 299-357) and the median relapse-free survival (RFS) was 136 months (95% CI 109-163). Pts treated with cladribine or pentostatin in first line had a statistically significant better RFS than pts treated with 'other' treatments (log rank test, p < 0.001). The 10-year cumulative incidence of relapse was 39% (95% CI 33-46). Pts who received treatments other than PNA in first line had a higher risk of relapse (Gray's test, p < 0.001). For pts receiving PNA in first and second lines, there was no difference in outcomes between those who switched PNA and those who did not. In this cohort, we observed 68 second malignancies during the follow-up: 49 solid cancers (most prevalent: prostate and non-melanoma skin cancers) and 19 hematological malignancies (most prevalent: monoclonal gammopathy of undetermined significance (MGUS) and myelodysplastic syndromes (MDS)). The median onset of second cancer, second solid cancer and second hematological malignancy from HCL diagnosis was 81 months, 99 months and 78 months, respectively. The median age at diagnosis of cancer, solid cancer and hematological malignancy was 70, 69 and 77 years old, respectively. Considering death as a competing risk, the 10-year cumulative incidence of cancer, solid cancer and hematological malignancy was 15% (95% CI 11-19), 11% (95% CI 7.2-15), and 5.0% (95% CI 2.8-8.2), respectively. In multivariate analyses, IFN treatment was associated with a decreased risk for all cancers (Fine and Gray regression model, subdistribution Hazard Ratio (sdHR) 0.53 (95% CI 0.29-0.97); p = 0.038), a familial history of cancer was a risk factor for solid cancers (sdHR 2.12 (95% CI 1.15-3.91); p = 0.017), a personal history of cancer was a risk factor for hematological malignancies (sdHR 3.47 (95% CI 1.14-10.55); p = 0.028). Even after excluding non-melanoma skin cancers and MGUS, there was an excess of cancers (SIR = 2.22), solid cancers (SIR = 1.81) and hematological malignancies (SIR = 6.67). Conclusions: In this updated real-world retrospective cohort with a long follow-up and most pts treated with PNA, we highlighted the importance and the excess of second cancers in HCL patients, in particular hematological malignancies. Figure Disclosures Paillassa: Janssen: Other: Bibliography board with young hematologists. Thieblemont:Roche: Honoraria, Research Funding; Gilead: Honoraria; Novartis: Honoraria; Kyte: Honoraria; Janssen: Honoraria; Celgene: Honoraria; Cellectis: Membership on an entity's Board of Directors or advisory committees. Hermine:AB Science: Membership on an entity's Board of Directors or advisory committees. Feugier:janssen: Honoraria, Research Funding, Speakers Bureau; gilead: Honoraria, Research Funding, Speakers Bureau; roche: Honoraria, Research Funding, Speakers Bureau; abbvie: Honoraria, Research Funding, Speakers Bureau. Troussard:Innate Pharma: Consultancy, Membership on an entity's Board of Directors or advisory committees; Abbvie: Consultancy, Membership on an entity's Board of Directors or advisory committees; Gilead: Consultancy, Membership on an entity's Board of Directors or advisory committees; Roche: Other: Research Support; Sysmex: Other: Research Support.

Subtype Distribution, Clinical Features, and Survival in B-cell Chronic Lymphoproliferative Disorders in China: A Review of 1592 Cases

BackgroundB-cell chronic lymphoproliferative disorders (B-CLPDs) are characterized by the sustained accumulation of monoclonal B cells. Limited studies have systematically described the clinical features and outcomes of the whole patient group, especially in Eastern populations. Patients and MethodsA total of 1592 patients with newly diagnosed B-CLPD were enrolled. Chronic lymphocytic leukemia (CLL) accounted for 39%, and Waldenström macroglobulinemia (WM), leukemic marginal zone lymphoma, follicular lymphoma (FL), and mantle cell lymphoma (MCL) constituted 13%, 13%, 9%, and 8% of cases, respectively. ResultsThe median age at diagnosis was 58 years, and the male/female ratio was 1.8:1. The 17p and 11q deletions were most common in MCL (36% and 17%, respectively), and 13q deletion and trisomy 12 were most frequent in CLL (35% and 21%, respectively). Patients with leukemic MCL had significantly worse survival than that of patients with other disease entities, with a 3-year overall survival (OS) of 58%, followed by 68.2% for WM/lymphoplasmacytic lymphoma. Those with CLL, leukemic marginal zone lymphoma, and FL had relatively favorable outcomes, with a 5-year OS > 80%. The survival of patients with B-CLPDs has improved over time with the emergence of novel drugs (3-year OS improvement from 82.1% to 92.2%). The improvement in survival mainly resulted from improvement among patients with MCL, WM/lymphoplasmacytic lymphoma, and FL. On multivariate analysis, only hemoglobin, lactate dehydrogenase, and 17p deletion were independently associated with survival (hazard ratio, 1.6, 2.0, and 3.1, respectively). ConclusionsComprehensive analysis of the clinical characteristics, immunophenotypic profiles, and cytogenetic features can be helpful in the differential diagnosis, especially for patients without a non–bone marrow biopsy specimen available. Universal prognostic factors could help with the early detection of high-risk patients and stratification for risk-adapted therapy.

A decision-tree approach for the differential diagnosis of chronic lymphoid leukemias and peripheral B-cell lymphomas

Background and ObjectiveHere we propose a decision-tree approach for the differential diagnosis of distinct WHO categories B-cell chronic lymphoproliferative disorders using flow cytometry data. Flow cytometry is the preferred method for the immunophenotypic characterization of leukemia and lymphoma, being able to process and register multiparametric data about tens of thousands of cells per second. MethodsThe proposed decision-tree is composed by logistic function nodes that branch throughout the tree into sets of (possible) distinct leukemia/lymphoma diagnoses. To avoid overfitting, regularization via the Lasso algorithm was used. The code can be run online at https://codeocean.com/2018/03/08/a-decision-tree-approach-for-the-differential-diagnosis-of-chronic-lymphoid-leukemias-and-peripheral-b-cell-lymphomas/ or downloaded from https://github.com/lauramoraes/bioinformatics-sourcecode to be executed in Matlab. ResultsThe proposed approach was validated in diagnostic peripheral blood and bone marrow samples from 283 mature lymphoid leukemias/lymphomas patients. The proposed approach achieved 95% correctness in the cross-validation test phase (100% in-sample), 61% giving a single diagnosis and 34% (possible) multiple disease diagnoses. Similar results were obtained in an out-of-sample validation dataset. The generated tree reached the final diagnoses after up to seven decision nodes. ConclusionsHere we propose a decision-tree approach for the differential diagnosis of mature lymphoid leukemias/lymphomas which proved to be accurate during out-of-sample validation. The full process is accomplished through seven binary transparent decision nodes.

CD43 and CD49d from the B-Cell Chronic Lymphoproliferative Disorders Diagnostic Panel Are Useful to Detect Erythroid Dysplasia.

Despite bone marrow (BM) immunophenotyping by flow cytometry has progressively been recognized as an important tool for the diagnosis of myelodysplastic syndromes (MDS), the sparse knowledge about normal erythroid maturation and the lack of markers for erythroid characterization is a major shortcoming. Here, we analyzed the expression of CD43 and CD49d, two markers included in the diagnostic panel for B-cell chronic lymphoproliferative disorders (B-CLPD), in the CD34+ compartment of normal BM and along the normal and dysplastic erythroid maturation. For this, 13 normal BM aspirates and 18 BM aspirates from MDS patients were studied by flow cytometry. Normal BM presented a higher expression of CD43 and CD49d among CD34+ erythroid precursors, compared to CD34+ cells committed to the remaining hematopoietic cell lineages. CD43 expression progressively decreased along the normal erythroid maturation, whereas CD49d levels increased from Stage I to Stage II, were maintained in Stages II and III, and then decreased until the last stage of maturation. In MDS, the expression of CD43 and CD49d followed a similar pattern, but with decreased expression levels for both markers, observed in all erythroid maturation stages (P < 0.05). Our results point to the usefulness of CD43 and CD49d, two markers commonly present in B-CLPD diagnosis panels, in the identification of dysplastic phenotypic features in the erythroid lineage. This allows a feasible and inexpensive way to identify patients who would benefit from a more extensive study to evaluate the presence of MDS, during the processing of suspected B-CLPD samples. © 2019 International Clinical Cytometry Society.

PB2112 COEXISTENCE OF MONOCLONAL B‐ CELL POPULATION IN PATIENTS WITH NEWLY DIAGNOSED MULTIPLE MYELOMA; INCIDENCE, IMMUNOPHENOTYPING AND CLINICAL CHARACTERIZATION

Background:Clustering of B‐cell malignancies in families, and recently described genetic correlations between multiple myeloma (MM) and chronic lymphocytic leukemia (CLL) (Went M et al; Blood Cancer Journal 2018) suggest shared biological pathways influencing the development of these malignancies. Since we began applying multiparameter flow cytometry (FACS) analysis of 8‐color panel for plasma cell dyscrasia (PCD), we observed several cases of newly diagnosed multiple myeloma (NDMM) with a co‐existing monoclonal B‐cell clone. Herein, we retrospectively review the incidence of this phenomenon and describe phenotypic and clinical characteristics.Aims:To investigate patients with NDMM and a co‐existing monoclonal B‐cell population.MethodsHospital records of all consecutive patients who underwent FACS analysis of bone marrow aspirates between January 2016 and September 2018 were reviewed and NDMM, active or smoldering, according to IMWG criteria, were identified. Cases of Waldenstrom's macroglobulinemia and lymphoplasmacytic lymphoma were excluded. FACS analysis included a PCD panel comprising CD45, CD138, CD38, CD27, CD56, CD19, and CytoKappa/Lambda. Cases that were found to have a monoclonal B‐cell population were further evaluated by a lymphoid screening tube (LST) and a B‐cell chronic lymphoproliferative disorders (BCLD) panel as proposed by EuroFlow™. Demographics, clinical characteristics, and outcomes of patients were documented from the hospital records.Results:Out of 160 consecutive FACS analyses of patients with NDMM, 13 cases (8.1%) had a coexisting monoclonal B‐cell population. In most cases, the monoclonal B‐cells had a CLL‐like phenotype (n = 8, 61.5%), whereas in the rest they were CD5‐negative (n = 5, 38.5%), with mean percentage of the monoclonal B‐cells of 5.7% (±1.7) out of all leukocytes. Patients with MM and co‐existing monoclonal B‐cells had a mean age of 72 years (range: 46‐86), 6 had smoldering and 7 had active myeloma and none of them had lymphadenopathy or splenomegaly. Involved immunoglobulin was IgG (n = 7), IgA (n = 3), IgM (n = 1) and undetected/unavailable (n = 2). PCD involved light chain was Kappa in 9 and Lambda in 4 patients. Involved free light chain level median was 332 mg/L (range 34‐9310) and M‐SPIKE median was 2.3 g/dL (range 0‐8.1). Plasma cells light chain subtype was concordant with that of the monoclonal B‐cells in 8 patients (61.5%). Hemoglobin median was 11.8 g/dL (range: 7.4‐14), absolute lymphocyte counts median was 1.7 × 109/L (1.1‐4.3). CRAB symptoms included lytic or focal lesions (n = 5), anemia (n = 5), renal failure (n = 1). Five of the 6 patients who received anti‐myeloma therapy responded. At a median follow‐up time of 9 months, none of the patients developed expansion of their B‐cell clone or progressed to an overt clinical lymphoproliferative disorder and 11 (80%) patients remain alive.Summary/Conclusion:We describe for the first time presence of a B‐cell monoclonal population in 8.1% of patients with NDMM. The clone is mostly with a CLL‐like phenotype, and did not expand or require therapy. While this incidence is slightly higher than expected, it may still be merely a coincidence reflecting prevalence of monoclonal‐B cell lymphocytosis in older population; alternatively, it may be caused by shared genetic drivers. Further investigations, including longer follow‐up and correlative molecular studies, are required.

High-dose methylprednisolone with Rituximab and fresh frozen plasma in the treatment of six patients with B-cell lymphoproliferative disorders harboring TP53 abnormalities

目的探究大剂量甲泼尼龙联合利妥昔单抗及新鲜冰冻血浆（HDMP+RTX+FFP）治疗伴有TP53基因异常[TP53缺失和（或）突变]的B细胞慢性淋巴增殖性疾病（B-CLPD）的疗效。方法自2008年5月至2012年5月前瞻性入组6例伴有TP53基因异常的B-CLPD患者，给予最多6个周期的HDMP+RTX+FFP治疗并进行疗效评估和随访。结果6例B-CLPD患者中包括4例B-CLPD-不能分类型（B-CLPD-U），1例B细胞型幼淋巴细胞白血病（B-PLL）和1例套细胞淋巴瘤（MCL）。中位治疗3（2~6）个周期，4例获得完全缓解（CR），其中3例达微小残留病（MRD）阴性的CR；1例疾病稳定（SD），1例疾病进展（PD）。中位随访30（4~56）个月，2例无治疗反应（NR）的患者均较快进展、死亡。4例CR患者均无进展，全部存活。治疗后患者的骨髓造血功能显著改善，但IgA、IgG及IgM水平明显下降。所有患者对此方案都表现出了良好的耐受性。骨髓抑制发生率较低，常见不良事件为不超过3级的中性粒细胞减少及感染，均可控。结论HDMP+RTX+FFP是治疗伴有TP53基因异常B-CLPD患者有效且易耐受的治疗方案。

Characteristics of CD180 Expression and Its Diagnostic Value in B Cell Chronic Lymphoproliferative Disorders

To investigate the charactcristics of CD180 expression and differentiation diagnostic value in B cell chronic lymphoproliferative disorders (B-CLPD) through detecting the mean fluorescence intensity（MFI）of CD180 in different sub types of B-CLPD，using multiparameter flow cytometry (FCM). The CD180 MFI of malignant B cells in 178 patients with B-CLPD was detected by FCM. The level of CD180 MFI in various types of B-CLPD was compared to the normal control group. The level of CD180 MFI among sub-types of B-CLPD was also compared. (1) The expression levels of CD180 in B-CLPD was significantly lower as compared with the normal controls，except the spleen difuse red pulp lymphoma (SDRPL); (2) The CD180 MFI in chronic lymphocytic leukemia sCLL) was significantly lower as compared with other B-CLPD cells; (3) CD180 ware significantly overexpressed in HCL compared with MCL，LPL，and MZL (P <0.05); (4) In the spleen-derived B-CLPD，such as SMZL，HCL, HCL variation and SDRPL，the expression of CD180 has significant difference between lymphomas with or without villous. Utilizing the multiparameter flow cytometry for defecting expression of CD180 and other immunological markers can more efficiently distinguish the subtypes of B-CLPD.

The Role of Molacular Genetic Examination in Differential Diagnosis and Prognostic Prediction in Patints with B-Cell Chronic Lymphoproliferative Disorders: A Restrospective Study of 1592 Patients

Background: B-cell chronic lymphoproliferative disorders (BCLPD) are a heterogeneous group of mature B-cell malignancies characterized by monoclonal proliferation in the bone marrow (BM) or peripheral blood (PB). The overlapped clinical features and the lack of individual diagnostic markers make diagnosis challenging, especially in patients who have difficulty to get adequate non-BM biopsy specimens or patients manifest as leukemic onset without nodes-involvement. It is necessary to explore novel methods to make precise diagnostic classification and prognostic prediction through simply PB/BM examination.Methods and results: A total of 1592 patients with newly diagnosed BCLPD were enrolled from January 1998 to December 2016, of which chronic lymphocytic leukemia (CLL) accounted for 39%, leukemic marginal zone lymphoma (l-MZL)occupied 13%, and Waldenstrom's macroglobulinemia/ lymphoplasmacytic lymphoma (LPL/WM), leukemic follicular lymphoma (l-FL) and mantle cell lymphoma (l-MCL) were 13%, 9% and 8%, respectively. There are 14% patients failed to fit into any of the categories and remaining unclassified which defined as BCLPDU. The median age at diagnosis was 58 years old. The male/female ratio of all BCLPD patients was 1.84:1. Male predominance was pronounced in WM/LPL, l-MCL and HCL with a M/F ratio of 2.5-2.7, while with balance gender in l-MZL and l-FL.FISH analyses were performed in 1234 patients. 17p and 11q deletion were most common in l-MCL (36% and 17%), while 13q deletion and trisomy 12 were most frequent in CLL (35% and 21%). Patients with 17p deletion showed significantly adverse survival in CLL and l-MCL (p<0.05), but the phenomenon was not found in other types of BCLPD. Instead, complex karyotype showed better predictive capability of worse survival than 17p deletion in patients with l-FL, l-MZL and BCLPDU.A total of 665 patients had been successfully amplified the IGHV-IGHD-IGHJ rearrangements. Based on a cut-off value of 98% homology, the IGHV mutated proportion in patients with l-MCL was 48.6%, which was higher than other BCLPDs, following CLL 33.3% and l-MZL 28.6%, while most of WM and l-FL were unmutated. The most frequent rearrangement in l-MZL were IGHV1-2*01, which was seldom found in other subtypes. Also, DH2-2 and VH3-7 were mainly found in CLL and l-MCL respectively. Besides, we have identified several stereotyped IGH CDR3 which were specific for the certain BCLPD subtype. About 23% unclassified patients could acquire further diagnosis according to the special mutation status, VDJ rearrangements or stereotyped CDR3 sequencing.With a median follow-up of 50.3 months, 522 patients (38.0%) had disease progression or died. Patients with lMCL have a significant worse survival than other entities, with a 3-year overall survival (OS) rate of 58%. Following , patients with WM/LPL had relatively poorer prognosis with a 5-year OS rate of 68.2%, while the prognosis for l-MZL, l-FL and CLL were relatively favourable, with a 5-year OS of 80%.The OS of the B-CLPD had improved over time due to the emergence of immunotherapy and novel (3-year OS improved from 82.1% to 92.2%). The improvement of l-MCL and WM was obvious but not for CLL and l-MZL.Conclusion: Non-BM biopsy specimens are not available for part of BCLPD patients. Overlapped magnification of BM/PB brings on difficulties in differential diagnosis. Besides morphology and immunophenotype, comprehensive analysis of FISH, karyotype, IGHV, MYD88 or BRAF mutation and other molecular examination might contribute to further differential diagnosis and prognostic prediction. Moreover, it might provide advice for the initial time and strategy of treatment. DisclosuresNo relevant conflicts of interest to declare.

CD200 is a useful marker in the diagnosis of chronic lymphocytic leukemia.

The diagnosis of CLL is supported by a typical morphology and immunophenotype and usually does not present difficulties. Nevertheless, some patients with CLL can show an atypical phenotype, this raising the possibility of a lymphoproliferative disorder other than CLL. It has been recently shown that the expression of CD200 could be a rather consistent marker for CLL. The expression of CD200 was investigated in 120 consecutive patients with B-cell chronic lymphoproliferative disorders (B-CLPD) (65 cases diagnosed as typical CLL, 16 atypical CLL, and 39 non-CLL before entering the study) by using multiparametric flow cytometry with four color combinations. CD200 was analyzed as percentage of positive cells (≥30%) and MFIR expression. ROC curves were used to determine the cut-off for the CD200 MFIR. Matutes score (MS) was used as comparator. All 81 (100%) patients classified as CLL and 25 of 39 (64.1%) classified as non-CLL expressed high CD200 expression (≥30%). CD200 expression showed a high sensitivity (100%) and a low specificity (35.9%), and the accuracy was similar to that of Matutes score markers (range: 79.2%-86.7%); except SmIg that was 59.1%. The addition of CD200 to the Matutes score correctly identified 74 of 81 (91.4%) CLL cases including 9 of 16 atypical CLL cases. As per non CLL cases, 37 of 39 (94.9%) were correctly diagnosed by the modified system. Altogether, CD200 improved the diagnostic accuracy of Matutes score from 86.7% to 92.5% (P < .01). These results show that CD200 is a valuable, albeit not specific, CLL diagnostic marker. © 2018 International Clinical Cytometry Society.

B-cell chronic lymphoproliferative disorders: a clinical analysis of 40 cases

Objective To investigate the clinical features, treatment and prognosis of the patients with B-cell chronic lymphoproliferative disorders (B-CLPD). Methods The data of 40 patients with B-CLPD in the Third People's Hospital of Kunshan from September 2010 to June 2017 were retrospectively analyzed, including clinical features, laboratory inspections, immunophenotyping, genetics and molecules results, therapeutic regimens, evaluation of curative effect and disease outcome. Results There were 29 male and 11 female patients in 40 B-CLPD patients, with a median age of 71.5 years old (47-88 years old). The percentage of chronic lymphocytic leukemia (CLL) was 57.5% (23/40), monoclonal B lymphocytosis was 10.0% (4/40), Waldenstrom macroglobulinemia was 15.0% (6/40), marginal/splenic marginal zone lymphoma was 12.5% (5/40), and mantle cell lymphoma was 5.0% (2/40). The immunophenotyping of the whole patients had the expressions of CD19, and surface immunoglobulin light chain in cytomembrane of 37 patients had a restrictive expression. All CLL patients presented the expressions of CD5 and CD23, while the other types of B-CLPD expressed various level of CD20, CD22, CD10, CD5, FCM-7. Twenty-six patients received chemotherapies including purine analogue, anthracyclines, alkylating agents and hormone. The overall response rate (complete remission plus partial remission) was 69.2% (18/26). The complete remission rate was 15.4% (4/26), which only occurred in the cohort of CLL patients who received the regimen containing fludarabine. The median follow up time of 26 patients who received medical treatment was 42.8 months (0.5-82.0 months), not reaching the median survival time. Conclusions The clinical features of B-CLPD are various, which requires comprehensive analysis of clinical data, including medical history, laboratory findings, imageological examination, cell morphology, immunophenotyping, genetics as well as molecular biology. The choice of the treatment should take the individualized situation into consideration. Key words: B-cell chronic lymphoproliferative disease; Clinical features; Diagnosis; Treatment

Albumin-to-Fibrinogen Ratio as an Independent Prognostic Parameter in Untreated Chronic Lymphocytic Leukemia: A Retrospective Study of 191 Cases.

PurposeChronic lymphocytic leukemia (CLL) is one of the most frequent type of B-cell chronic lymphoproliferative disorders and chronic inflammation takes part in the development of CLL. However, there has been no valid immune biomarker to predict the prognosis of untreated CLL patients.Materials and MethodsIn this retrospective study, we analyzed the clinical correlations and prognostic value of albumin-to-fibrinogen ratio (AFR) detected at diagnosis in 191 CLL patients.ResultsThe cut-off value of AFR was 9.7 calculated by X-tile. Patients who were more than 65 years old were often accompanied by low level of AFR (p < 0.001). Survival analysis showed that patients with low level of AFR had shorter overall survival (OS) than patients with high level of AFR (p < 0.001). Multivariate analysis illustrated that AFR had a negative impact on OS (p=0.003) and was independent of parameters involved in CLL international prognostic index and other prognostic markers such as CD38 and ZAP-70.ConclusionThese data provide a comprehensive view of AFR and shows that AFR at diagnosis is an adverse prognostic factor in untreated CLL patients.

Del17p does not always significantly influence the survival of B-cell chronic lymphoproliferative disorders.

B-cell chronic lymphoproliferative disorders (B-CLPD) comprise several entities with indolent clinical manifestations but heterogeneous survival. Cytogenetic aberrations are now the standard prognostic predictors in chronic lymphocytic leukemia (CLL) but have been less investigated in other subtypes of B-CLPD. In this study, we detected cytogenetic aberrations by fluorescence in situ hybridization (FISH) in 875 B-CLPD patients, based on a panel probes locating at 13q14, 11q22, 17p13 and CEP12. We identified del17p acted as the independent adverse cytogenetic predictor for overall survival (OS) in CLL. Del13q, del11q and del17p were adverse factors for OS in Waldenström's macroglobulinemia in the univariate analysis but lost their role in the multivariate analysis. Trisomy 12 acted as an independent poor factor for both marginal zone lymphoma (MZL) and unclassified B-CLPD (BCLPD-U) subtype. Del17p did not impact survival in MZL and BCLPD-U patients. These contrasting results indicate different roles of the same cytogenetic aberrations in the pathogenesis of each B-CLPD subtype. As del17p contributed to the poorest survival in CLL and desired extraordinary treatment strategy, the imitation of CLL strategy to other B-CLPD with del17p should be carefully advocated based on this study.

Spectrum and immunophenotyping of 653 patients with B-cell chronic lymphoproliferative disorders in China: A single-centre analysis.

The incidence of B-cell chronic lymphoproliferative disorders (B-CLPDs) is significantly lower in China than that in western countries. There have been studies involving small cohorts with conflicting results regarding the spectrum of B-CLPDs in China, and the types and immunophenotyping of B-CLPDs in China remain largely unexplored. We conducted a retrospective analysis of 653 cases of B-CLPDs seen in our centre from 2011 to 2015. Four-colour flow cytometry was used to determine the expression of each immunological marker, and the diagnostic values of the immunological markers were also investigated. Chronic lymphocytic leukaemia (CLL) was the most common type of B-CLPD, which was consistent with that in west countries. However, the proportions of CLL (55.9%), follicular lymphoma (2.6%), and hairy cell leukaemia (0.2%) were lower, while the proportion of lymphoplasmacytic lymphoma/WaldenstrÖm macroglobulinaemia (5.4%) was higher in China, as compared with western countries. With respect to immunophenotypic characteristics, CD23 (31.7%) was more frequently expressed in mantle cell lymphoma (MCL) in our cohort than that in western countries. Immunophenotyping was useful in differentiating MCL from CLL or B-cell prolymphocytic leukaemia and lymphoplasmacytic lymphoma/WaldenstrÖm macroglobulinaemia from splenic marginal zone lymphoma. CD200 was of better diagnostic performance (accuracy: 94.6%) in differentiating CLL from MCL compared with CD23 (accuracy: 93.3%). Some cases of B-CPLDs, however, had no definite diagnoses, which were diagnosed as CD5+ B-CPLDs unclassified (7.7%) and CD5- B-CPLDs unclassified (15.8%). This is the largest study that systematically explores the spectrum and immunophenotyping of B-CLPDs in Asia, confirming that spectrum of B-CLPDs in China was different from that in western countries. The immunophenotypic features of B-CLPDs were similar between China and western countries, although a few disparities exist. Cases with no definite diagnoses warrant further studies in the future.

The prognostic significance of 13q deletions of different sizes in patients with B-cell chronic lymphoproliferative disorders: a retrospective study.

Patients with chronic lymphocytic leukemia (CLL) with 13q deletion as the sole cytogenetic abnormality usually have a favorable outcome, but the frequency of the 13q14 deletion and its impact on the outcome of other B-cell chronic lymphoproliferative disorders (BCLPDs) remain unclear. To further characterize this aberration, we investigated the prognostic significance of 13q deletion in 541 patients with BCLPDs. We performed fluorescence in situ hybridization (FISH) studies with 13q locus-specific LSI-D13S25 and LSI-RB1 probes. 52.1% of the patients with CLL harbored 13q deletion, which was significantly higher than that of other BCLPDs (p<0.001). The size of 13q deletion was heterogeneous in both CLL and other BCLPDs. However, the distribution of cases with different deletion sizes showed no significant difference between the two groups. Whereas 13q deletion was a favorable prognostic factor in CLL, a large deletion of 13q was associated with poor prognosis in terms of time to first therapy (p=0.020), progression-free survival (p=0.05) and overall survival (p=0.002) in BCLPD cases other than CLL. In conclusion, the deletion of 13q varied in size both in CLL and in other BCLPDs and adversely influenced the prognosis of patients with other BCLPDs.

B-cell chronic lymphoproliferative disorders misdiagnosed successively as autoimmune hemolytic anemia and pure red-cell aplasia: report of one case and review of literature

目的 探讨慢性B淋巴细胞增殖性疾病（B-CLPD）的临床及实验室特征。 方法 报道1例先后以自身免疫性溶血性贫血（AIHA）及纯红细胞再生障碍（PRCA）为表现的B-CLPD患者的临床资料及诊治经过，结合文献复习，探讨B-CLPD的临床及实验室特征及诊断。 结果 B-CLPD常见临床表现为淋巴结、脾大以及骨髓、外周血中淋巴细胞增多等淋巴增殖表现。但极少数患者仅以血细胞减少为主要表现，而无典型淋巴细胞增殖特征，容易导致误诊。明确诊断及分型需要综合分析外周血及骨髓细胞涂片、骨髓病检、免疫分型、细胞遗传学及分子学检查。 结论 对于AIHA及PRCA患者，即使外周血淋巴细胞无增多，亦应全面完善骨髓涂片、淋巴细胞免疫表型分析、IGH基因重排以及特异性的遗传分子学相关检查，除外B-CLPD，减少误诊。

Expression of serum monoclonal immunoglobulins in B-cell chronic lymphoproliferative disorders and their significances

Objective To investigate the incidence of serum monoclonal immunoglobulins (McIg) in B-cell chronic lymphoproliferative disorders (B-CLPD) and the clinical significance of McIg in B-CLPD and its possible sources. Methods A total of 1 147 patients with B-CLPD treated from May 2006 to May 2015 were enrolled into this retrospective study. The incidence of McIg and the relationship between McIg and prognostic factors in patients with B-CLPD were analyzed. Results Out of 1 147 B-CLPD patients, there were 164 patients with lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM), and among them, McIg was detected in 140 cases (85.4%). In the remaining 983 patients with B-CLPD, monoclonal Ig was detected in 50 (5.1%) patients. Most of McIg in 2 groups were IgM paraprotein. The levels of IgM paraprotein of the LPL/WM group, non-LPL/WM group and McIg-negative patients were (48.88 ± 33.42) g/L, (27.9 ± 15.23) g/L and (2.75±1.21) g/L, respectively, the difference was statistical significance (P=0.000); the level of IgM paraprotein in LPL/WM group was significantly higher than that in non-LPL/WM group (P=0.000). The level of paraprotein decreased significantly when the patients got complete response after therapy (P=0.001, 0.048, respectively). The incidence of serum McIg was higher in the group with complex karyotype (P=0.016) and with high level of β2-microglobulin (β2-MG) (P=0.001). In the 47 non-LPL/WM patients with positive McIg, serum McIg in 38 (80.9%) patients were expressed in a pattern consistent with the distribution of tumor cells (P<0.005). Most of the light chain subtype of the McIg were consistent with the light chain subtype of the membrane immunoglobulin on the tumor cells. Conclusions Some non-LPL/WM B-CLPD patients also have serum McIg, and it could have certain relevance with the prognosis of B-CLPD. Moreover, the McIg may be secreted by tumor cells or those derived from the same progenitor cells with tumor cells. Key words: Lymphoproliferative disorders; Immunoglobulin; Prognosis

Prognostic Significance of Serum LDH in B Cell Chronic Lymphoproliferative Disorders: A Single-Institution Study of 829 Cases in China

Background: B-cell chronic lymphoproliferative disorders (B-CLPD) comprise several indolent lymphoma subtypes with similar characteristics but also difference and heterogeneity. Several prognostic systems had been established for separate subtypes, such as Rai/Binet stage system for CLL, FLIPI for follicular lymphoma. Here, we want to explore the prognostic role of some common clinical characteristics in each lymphoma subtype, such as serum LDH.Materials and methods: The clinical data of 829 patients with B-CLPD including clinical features, LDH expression level, cytogenetic abnormalities and therapeutic outcome were analyzed retrospectively in department of lymphoma & myeloma, Institute of hematology & Blood disease hospital from April 1990 to August 2013, and the prognosis of B-CLPD patients with different expression levels of LDH was compared, including overall survival (OS) and progression-free survival (PFS) rates.Results: 829 patients were divided into three groups, including 426 (51.4%) patients with chronic lymphocytic lymphoma (CLL), 307 (37.0%) patients with non-CLL B-CLPD and 96 (11.6%) patients with BLPD-U. Elevated serum LDH at diagnosis was detected in 112 (26.3%), 85 (27.7%) and 23 (24.9%) of CLL, non-CLL B-CLPD and BLPD-U group, respectively. Patients with elevated LDH B-CLPD had poorer 5-year overall survival rate (CLL: 63.8% vs 83.0%; non-CLL B-CLPD: 62.2% vs 78.1%; BLPD-U: 47.4% vs 88.6%) and 5-year progression-free survival rate (CLL: 52.0% vs 78.2%; non-CLL B-CLPD: 40.8% vs 75.6%; BLPD-U: 48.9% vs 89.5%) than LDH normal patients. Univariate analysis showed that LDH expression, hemoglobin level, platelet count, B symptoms, P53 deletion were prognostic factors for CLL patient survival, while LDH expression, platelet count, albumin level affected non-CLL B-CLPD patient survival significantly and LDH expression, albumin level, ECOG PS scores for BLPD-U patient survival. Multivariate analysis showed that elevated LDH was an independent significant prognostic factor for overall survival (P<.001) and progression-free survival (P<.001) in all three groups.Conclusions: Based on these data, we conclude that elevated serum LDH at diagnosis is an unfavorable prognostic factor in patients with B-CLPD. DisclosuresNo relevant conflicts of interest to declare.

Bendamustine and Rituximab, as First Line Treatment, in Intermediate, High Risk Splenic Marginal Zone Lymphomas of Elderly Patients.

BackgroundSplenic marginal zone lymphoma (SMZL) is a chronic B-cell lymphoproliferative disorder, comprising less than 2% of non-Hodgkin’s lymphomas, and affecting mainly middle-aged and elderly patients with a median survival of >10 years. The typical clinical features of SMZL include splenomegaly. Treatment should be patient-tailored and can range from a ‘watchful waiting’ approach for asymptomatic patients without cytopenias to surgery, localized radiation therapy or immuno/chemotherapies. Recently, the combination of rituximab and Bendamustine (R-Benda) has been defined as highly active in patients with follicular lymphomas, but little is known about the efficacy of R-Benda in SMZL.Aim of the studyThe purpose of this retrospective study was to report our experience on the efficacy of R-Benda as first line treatment in 23 consecutive elderly SMZL patients.ResultsAll patients had a complete resolution of splenomegaly along with restoration of their blood counts. Nineteen patients (83%) achieved a complete response (CR) to therapy; three patients (13%) achieved a partial response (PR).Ten patients (43%) obtained molecular remission. Toxicities were mild and mainly haematological and result in dose reductions for fourteen patients.ConclusionsOur data suggest a high activity and good tolerance of R-Benda, despite dose reduction due to potential toxicity.

Autoimmunity and lymphoproliferation markers in naïve HCV-RNA positive patients without clinical evidences of autoimmune/lymphoproliferative disorders

BackgroundHCV can lead to both chronic liver disease and B-cell lymphoproliferative disorders. A strong association exists between HCV and mixed cryoglobulinaemia (MC). MethodsAnti-nuclear antibodies (ANA), rheumatoid factor Ig-G (RF-IgG), free light chain κ and λ (FLC-κ, FLC-λ) levels and κ/λ ratio were evaluated in 50/420 subjects unexpectedly resulted anti-HCV positive after routine screenings for non-hepathological procedures. ResultsThree/fifty patients had HCV-RNA undetectable in the serum and were excluded from the analysis. Thirty-nine/fifty patients had laboratory evidence of circulating cryoglobulins without liver disease and MC-related symptoms. Among them, 17 resulted ANA-positive.The mean cryocrit was higher in ANA-positive patients, while no other demographic/clinical differences were observed between the groups. Significantly higher levels of RF-IgG were observed in ANA-positive vs ANA-negative patients. κ and λ FLC were higher in ANA-positive patients.A ROC analysis, based on ANA-positivity vs ANA-negativity, confirmed a high sensitivity and specificity of RF-IgG test. ConclusionsPublished data concerning MC come mostly from symptomatic vasculitis. We analyzed HCV-patients without MC symptoms, founding cryoglobulins in the majority of them.The increased levels of FR-IgG and FLC in CGs-ANA-positive patients, suggest these test could be used to identify a state of silent autoimmune and/or lymphoproliferative condition before the transition to a frank disease in naïve HCV-patients without symptoms of extrahepatic manifestations.

Low expression of CD200 predicts shorter time-to-treatment in chronic lymphocytic leukemia.

CD200, formerly known as OX-2, is a type I glycoprotein that is expressed on a variety of cell types. CD200 has been shown to be overexpressed in chronic lymphocytic leukemia (CLL). Although previous studies have confirmed the diagnostic value of CD200 in differentiating CLL from to other B-cell chronic lymphoproliferative disorders especially mantle cell lymphoma, whether CD200 has prognostic significance in CLL remains to be determined. We evaluated the mean fluorescence intensity (MFI) of CD200 in 307 consecutive, untreated patients with CLL in our center using flow cytometry. Using a CD200 MFI cutoff of 189.5, these cases could be divided into two groups. Patients with lower CD200 MFI (< 189.5) had a significantly shorter time-to-treatment (TTT) than those with higher CD200 MFI (≥ 189.5) (median TTT: 2 months vs 28 months, p = 0.0008). However, the effect of CD200 MFI on overall survival was not significant (CD200 MFI < 189.5: undefined vs CD200 MFI ≥ 189.5: undefined, P = 0.2379). In subgroup analysis, CD200 MFI retained its prognostic value in patients with favourable characteristics such as Binet stage A disease, mutated IGHV status, normal TP53 or negative CD38 expression. In conclusion, our study identified CD200 MFI as a potential prognostic factor in CLL.