Cerebrospinal Fluid Immunophenotyping in B-Cell Lymphomas: A Single Centre Experience on 99 Samples.

Abstract Central nervous system involvement in B-cell lymphomas is almost always lethal. Recognized risk factors include Burkitt histology and high LDH level, multiple extranodal sites or an age-adjusted International Prognostic Index of > 1 in the case of diffuse large B-cell lymphoma (DLBCL) (van Besien et al. Blood 1998, Haioun, Ann Oncol 2000). Standard diagnostic approaches such as imaging (MRI, CT) and cytological study of the cerebrospinal fluid (CSF) are flawed with a large number of false negative results. Accordingly, currently used prophylactic approaches based on these uncertain criteria carry a risk of over- or undertreating patients. Lately, flow cytometry (FCM) has been shown to detect a high incidence of “occult” CNS disease in patients at risk (Hedge et al. Blood 2005), thus paving the way towards a more specific management of these cases. We performed an exploratory single-center study on 99 CSF samples from patients with B-cell lymphomas. FCM results did not influence the therapeutic strategy. CSF samples were processed as soon as possible following lumbar puncture (always within 3hrs). They were washed in saline buffer, counted in Trypan Blue (to evaluate cellularity & viability), and 1/3 was labelled with the following antibodies: CD45, CD3, CD14, CD19, CD5, and CD10. Two thirds of the remaining cells were either labelled with the same antibodies (in case paucicellularity did impede interpretation of the first labelling on one third of the sample), or with one of two complementary antibody combinations: CD20, CD22, CD19 or Kappa/Lambda/CD19 according to cellularity and clinical data. From July 2005 to April 2007, 99 samples from 87 consecutive patients were analyzed. Median age was 61 years and 64% patients had an IPI score of 3 or more. Forty six patients were devoid of neurological symptoms at the time of analysis. Forty had DLBCL, 6 Burkitt. Thirteen out of the 99 samples could not be analyzed because they were massively contaminated with blood, 7 because they showed non specific fluorescence, thus leaving 79 (80%) analyzable samples from 75 patients. Although some samples contained an excess of B-cells (22% with ≥ 5% and < 10% CD19+ cells, 22% with ≥10% CD19+ cells), none showed a clearly phenotypically abnormal B-cell population. On a clinical basis, all were treated with curative intent and 82% had meningeal prophylaxis. No patient developed CNS localization with a median follow-up of 15 months.Twenty-five samples were taken from 19 patients with neurological symptoms (cranial nerve/radicular palsies n=9, CNS tumors n=6, epiduritis n=4). Three cases showed a clearly phenotypically abnormal B-cell population (1 case of Burkitt lymphoma, 2 cases of lymphoplasmacytoid lymphoma), all being also positive by cytology.In conclusion, although preliminary, these results suggest that in our hand FCM appears not more sensitive that cytology in detecting CSF infiltration by neoplastic B-cells in aggressive B-cell lymphoma when performing a single exploratory lumbar puncture. A larger cohort of patients needs to be explored with standardization of FCM immunophenotyping technique in order to improve further diagnostic accuracy of CSF localization in lymphoma.

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Deregulation of CCND1 or BCL2 in the Immature B-Cell Stage Determines the Resulting Lymphoma Histology

Blood ◽

10.1182/blood.v120.21.416.416 ◽

2012 ◽

Vol 120 (21) ◽

pp. 416-416

Author(s):

Masaharu Tashima ◽

Momoko Nishikori ◽

Wataru Kishimoto ◽

Ryo Yamamoto ◽

Tomomi Sakai ◽

...

Keyword(s):

B Cells ◽

Mouse Model ◽

B Cell ◽

Tumor Cells ◽

Cell Population ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

B Cell Lymphomas ◽

Cell Stage ◽

Genetic Event

Abstract Abstract 416 In B-cell lymphomas, several chromosomal translocations are associated with specific histological subtypes. CCND1/IGH is detected in more than 90 % of the cases with mantle cell lymphoma (MCL), and BCL2/IGH is characteristically observed in follicular lymphoma and diffuse large B-cell lymphoma (DLBCL) of germinal center (GC) B-cell origin. Although these strong correlations are clinically recognized, their biological mechanism is not clearly explained so far. According to the results of translocation breakpoint mapping, both CCND1 and BCL2 translocations are considered to be generated by an error during physiological VDJ rearrangement of the IGH gene in the precursor-B cell stage. We hypothesized that the occurrence of these translocations in the immature B-cell stage, probably as an initial genetic event, should have a special impact on the determination of resulting lymphoma histology. We generated a mouse model mimicking human lymphoma with CCND1 or BCL2 translocation by lentivirally introducing these genes into Tp53+/− B6 mouse bone marrow cells and transplanting them to lethally irradiated wild-type B6 mice. In this model, CCND1 or BCL2 is expressed from immature to mature B cell stage under the control of the CD19 promoter, and subsequently mutations accumulate in the background of Tp53 haploinsufficiency. Both mice developed B-cell lymphomas several months after the transplantation, but their tumors showed some different features. CCND1-Tp53+/− mice developed B220lowCD5+CD23− tumors that expand into the B-cell area with sparing the GC, whereas BCL2-Tp53+/− mice preferentially developed B220highCD5−CD23+ tumors that tend to localize in the GC. Additionally, somatic hypermutation (SHM) analysis of the IGH gene of the tumor cells revealed obviously higher mutation frequency in the BCL2-Tp53+/− mice than in the CCND1-Tp53+/- mice (p<0.004). These results indicate that the primary gene deregulation of CCND1 and BCL2 determines the upcoming lymphoma of MCL-type and GC B-cell lymphoma-type, respectively. MCL has been postulated to be derived from naïve pre-GC B cells, with few SHM introduced in the immunoglobulin variable region. But the MCL-like tumor cells generated in CCND1-Tp53+/−mice seem to be originated from B-1a B cells, a distinct B cell population that does not enter GC and has few SHM in nature. There has been a debate whether B-1 cell population exists in humans, but it is recently proposed that the phenotype of human B-1 cells is CD20+CD27+CD43+CD70− by testing sort-purified B cell fractions for fundamental B-1 cell functions based on mouse studies (J Exp Med 2011;208:67–80). Interestingly, flow cytometric analysis of the human MCL tumor cells has shown that they mostly express this provisional B-1 cell phenotype, supporting the idea that human MCL is also derived from B-1 cells. Our mouse model precisely reproduces the link between CCND1 and BCL2 translocations and the resulting lymphoma subtypes in humans. It is assumed that these translocations trigger the cell expansion of different B-cell subgroups, which consequently leads to the development of lymphoma of distinct histology. Our findings provide new insights into the mechanism of lymphoma subtype determination. Disclosures: No relevant conflicts of interest to declare.

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Distinct roles for PARP-1 and PARP-2 in c-Myc-driven B-cell lymphoma in mice

Blood ◽

10.1182/blood.2021012805 ◽

2021 ◽

Author(s):

Miguel A Galindo-Campos ◽

Nura Lutfi ◽

Sarah Bonnin ◽

Carlos Martínez ◽

Talia Velasco-Hernandez ◽

...

Keyword(s):

Dna Damage ◽

B Cells ◽

B Cell ◽

Immune Escape ◽

Cell Lymphoma ◽

Tumour Progression ◽

Cancer Therapeutics ◽

B Cell Lymphoma ◽

B Cell Lymphomas ◽

Parp 1

Dysregulation of the c-Myc oncogene occurs in a wide variety of haematologic malignancies and its overexpression has been linked with aggressive tumour progression. Here, we show that Poly (ADP-ribose) polymerase (PARP)-1 and PARP-2 exert opposing influences on progression of c-Myc-driven B-cell lymphomas. PARP-1 and PARP-2 catalyse the synthesis and transfer of ADP-ribose units onto amino acid residues of acceptor proteins in response to DNA-strand breaks, playing a central role in the response to DNA damage. Accordingly, PARP inhibitors have emerged as promising new cancer therapeutics. However, the inhibitors currently available for clinical use are not able to discriminate between individual PARP proteins. We found that genetic deletion of PARP-2 prevents c-Myc-driven B-cell lymphomas, while PARP-1-deficiency accelerates lymphomagenesis in the Em-Myc mouse model of aggressive B-cell lymphoma. Loss of PARP-2 aggravates replication stress in pre-leukemic Em-Myc B cells resulting in accumulation of DNA damage and concomitant cell death that restricts the c-Myc-driven expansion of B cells, thereby providing protection against B-cell lymphoma. In contrast, PARP-1-deficiency induces a proinflammatory response, and an increase in regulatory T cells likely contributing to immune escape of B-cell lymphomas, resulting in an acceleration of lymphomagenesis. These findings pinpoint specific functions for PARP-1 and PARP-2 in c-Myc-driven lymphomagenesis with antagonistic consequences that may help inform the design of new PARP-centred therapeutic strategies with selective PARP-2 inhibition potentially representing a new therapeutic approach for the treatment of c-Myc-driven tumours.

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Profiles Of De Novo CD25-Positive Mature B-Cell Lymphomas

Blood ◽

10.1182/blood.v122.21.4308.4308 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4308-4308

Author(s):

Shin-ichiro Fujiwara ◽

Raine Tatara ◽

Kiyoshi Okazuka ◽

Iekuni Oh ◽

Ken Ohmine ◽

...

Keyword(s):

Flow Cytometry ◽

B Cells ◽

B Cell ◽

Germinal Center ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

High Expression ◽

B Cell Lymphomas ◽

Cns Involvement ◽

Cd25 Expression

Abstract Background Interleukin 2 (IL-2) is an important cytokine that controls the proliferation and differentiation of not only T- but also B-lymphocytes. Recently, we reported that CD25 (IL-2 receptor alpha chain, IL-2R) is expressed in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL), and high expression of CD25 in the two types of lymphoma is correlated with a poor prognosis following chemotherapy regimens containing rituximab (ASH annual meeting, 2011 118:2666, 2012 120:1543). We evaluated the clinical significance of CD25 expression in a larger series of different mature B-cell lymphomas (BCL). Patients and Methods Four hundred and thirty-seven newly diagnosed patients who were admitted to our hospital between 2002 and 2013 were retrospectively evaluated. Lymph node or related tissue biopsy samples of BCL were analyzed using flow cytometry, as follows: 182 patients, DLBCL; 92, FL; 48, chronic lymphocytic leukemia (CLL); 21, mantle cell lymphoma (MCL); 23, marginal zone lymphoma (MZL); 8, Burkitt lymphoma (BL); 18, B-cell lymphoma unclassifiable with features intermediate between BL and DLBCL (BL/DLBCL); 5, lymphoplasmacytic lymphoma (LPL); and 39, reactive lymphadenopathy with sufficient B-cells. CD25-positivity was defined as >20% of clonal B-cells in a gated region. Results CD25 expression in patients with MCL, CLL, MZL, and DLBCL was significantly higher than that in patients with reactive lymphadenopathy (P<0.001,<0.001, =0.019, and <0.001, respectively). BL and FL, which were derived from germinal center B-cells, did not express CD25. These results indicate that pre- or post- germinal center-derived B-cells, activated by IL-2/IL-2R signaling, may give rise to CD25+ BCL such as CD25+ MCL, CLL, MZL, and DLBCL. The highest median CD25 expression (41.5%) was observed in MCL. CD25 expression was higher in MCL than CD5+ BCL (CLL and CD5+ DLBCL) (median, 41.5 vs. 16.9%, respectively; P<0.001). With a cut-off value of 60% CD25-positivity, patients with CD25-high (>60%) MCL (n=9) were not treated with aggressive chemotherapy regimens such as Hyper-CVAD due to their age and characteristics, compared with those with CD25-low (<60%) MCL (n=12) (11.1 vs. 72.7%, respectively, P=0.021). In patients with CLL, the range of CD25 expression was wide (0.4-90.7%), and 29 patients (60%) showed CD25-positivity (CD25+ CLL). CD25+ CLL showed higher soluble IL-2R (sIL-2R) levels and an inferior overall survival (OS) than CD25- CLL (median sIL-2R, 2,195 vs. 706 U/ml P=0.047; 5-year OS, 62.7 vs. 100%; P=0.037). There was a significant correlation between levels of CD25 and sIL-2R (r=0.53, P=0.0053). It is clinically important to distinguish between DLBCL and BCL involving MYC oncogene rearrangement (BL and BL/DLBCL, MYC+ BCL). The former showed higher CD25 expression than the latter (median, 10.2 vs. 2.1%, respectively, P=0.04). The progression-free survival rate (PFS) after rituximab containing chemotherapy was inferior in patients with CD25+ DLBCL (n=72) than those with CD25- DLBCL (n=110) and MYC+ BCL (5-year PFS, 49 vs. 70.4, 66.3%, respectively). In patients with DLBCL, central nerve system (CNS) involvement was observed in 15 patients (7 at diagnosis and 8 at relapse). CD25+ DLBCL showed a higher frequency of CNS involvement than CD25– DLBCL (13.8 vs. 4.5%, respectively, P=0.049). Regarding MZL, CD25 was highly expressed in nodal MZL, but it showed a low expression in splenic MZL. Regarding the sites of extranodal MZL, CD25 expression was lower in the thyroid than at other sites (median, 5.1 vs. 21.2%, respectively, P=0.37). There were some differences between CD25+ (n=9) and CD25- (n=14) MZL concerning the presence of B symptoms (33.3 vs. 0%, respectively) and advanced stage (66.6 vs. 35.7%, respectively). Conclusion CD25 expression using flow cytometry can potentially provide diagnostic and prognostic implications on BCL patient. The high expression of CD25 in MCL and CLL suggests the possibility of targeted anti-CD25 immunotherapy. These findings may shed light on the role of CD25 expression in B-cell lymphomagenesis. Disclosures: No relevant conflicts of interest to declare.

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Persistent expression of the full genome of hepatitis C virus in B cells induces spontaneous development of B-cell lymphomas in vivo

Blood ◽

10.1182/blood-2010-05-283358 ◽

2010 ◽

Vol 116 (23) ◽

pp. 4926-4933 ◽

Cited By ~ 55

Author(s):

Yuri Kasama ◽

Satoshi Sekiguchi ◽

Makoto Saito ◽

Kousuke Tanaka ◽

Masaaki Satoh ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

B Cells ◽

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Hcv Infection ◽

Mouse Serum ◽

B Cell Lymphomas ◽

Α Subunit

AbstractExtrahepatic manifestations of hepatitis C virus (HCV) infection occur in 40%-70% of HCV-infected patients. B-cell non-Hodgkin lymphoma is a typical extrahepatic manifestation frequently associated with HCV infection. The mechanism by which HCV infection of B cells leads to lymphoma remains unclear. Here we established HCV transgenic mice that express the full HCV genome in B cells (RzCD19Cre mice) and observed a 25.0% incidence of diffuse large B-cell non-Hodgkin lymphomas (22.2% in males and 29.6% in females) within 600 days after birth. Expression levels of aspartate aminotransferase and alanine aminotransferase, as well as 32 different cytokines, chemokines and growth factors, were examined. The incidence of B-cell lymphoma was significantly correlated with only the level of soluble interleukin-2 receptor α subunit (sIL-2Rα) in RzCD19Cre mouse serum. All RzCD19Cre mice with substantially elevated serum sIL-2Rα levels (> 1000 pg/mL) developed B-cell lymphomas. Moreover, compared with tissues from control animals, the B-cell lymphoma tissues of RzCD19Cre mice expressed significantly higher levels of IL-2Rα. We show that the expression of HCV in B cells promotes non-Hodgkin–type diffuse B-cell lymphoma, and therefore, the RzCD19Cre mouse is a powerful model to study the mechanisms related to the development of HCV-associated B-cell lymphoma.

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A Novel NF-kB:NFAT Enhanceosome Synergistically Regulate CD154 Gene in Aggressive B Cell Lymphomas.

Blood ◽

10.1182/blood.v104.11.28.28 ◽

2004 ◽

Vol 104 (11) ◽

pp. 28-28 ◽

Cited By ~ 1

Author(s):

Lan Pham ◽

Archito Tamayo ◽

Linda Yoshimura ◽

Yen-Chiu Lin-Lee ◽

Richard J. Ford

Keyword(s):

Transcriptional Regulation ◽

B Cells ◽

B Cell ◽

Gene Transcription ◽

Promoter Activity ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Cd40 Ligand ◽

B Cell Lymphomas ◽

Incidence And Mortality

Abstract Non-Hodgkin’s Lymphoma (NHL) causes many deaths world wide, and is one of the few cancers that have a continual increase in incidence and mortality rates over the last few decades. Diffused large B cell lymphoma (DLBCL) is an aggressive B cell lymphoma that accounts approximately 40% of all NHL-B cells. We have previously shown that dysregulated CD40 ligand (CD40L/CD154) expression in DLBCL maintains lymphoma cell growth and cell survival. CD40 ligand is a member of the TNF superfamily of proteins that has a wide variety effects throughout the immune system and is critical for both cellular and humoral immunity. It was originally thought that CD154 expression was restricted to activated CD4+ T lymphocytes. However, expression of CD154 has also been demonstrated in non-lymphocytic leukocytes such as mast cells, basophils, eosinophils, dendritic cells and macrophages. More importantly, dysregulated CD154 expression has been noted in a number of diseases, including systemic lupus erythematosus (SLE), Alzheimer’s, and in B cell lymphomas. Although the functional analysis of CD154 has been extensive, little is known about the mechanisms controlling CD154 expression in activated T cells, let alone in normal and malignant B cells. In this study, we show that constitutive NF-kB and NFAT activation in aggressive lymphoma B cells directly interact and synergistically regulate CD154 gene expression. Immunoprecipitation and confocal analyses indicate that NFATc1 and NF-kB (p65 and c-rel) complex and colocalize in lymphoma B cells. Chromatin Immunoprecipitation (CHIP) assays analysis demonstrate that NFATc1, p65 and c-rel bind to the CD154 promoter, that is enhanced by HDAC inhibitor (TSA or SAHA) treatments. HDAC inhibitors also enhance CD154 promoter activity, providing evidence that CD154 transcriptional regulation involves histone acetylation. Over-expression studies show a synergistic effect by NFATc1 and NF-kB (c-rel) on the CD154 promoter. Promoter deletion and site direct mutagenesis studies on the CD154 promoter reveal that the transcriptional regulation of CD154 requires two sites, the distal kB site at −1180 and a proximal NFAT site at −250. These sites bind to both, NFAT and NF-kB proteins, as wells as the DNA architectural protein HMG-1, indicating that the CD154 promoter DNA may loop and form an enhanceosome-like complex. Studies using siRNA to p65, c-rel, or NFATc1 repress CD154 promoter activity, indicating that these transcription factors are necessary and sufficient for CD154 gene transcription. These findings strongly suggest a novel control mechanism for CD154 gene transcription, therefore, providing potential treatment modalities for lymphoma B cells as well as other disorders involving dysredulated CD154 expression.

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Inactivation Of BANK1 By a Novel IGH-Associated Translocation and 5’ Hypermethylation In B-Cell Lymphomas

Blood ◽

10.1182/blood.v122.21.2497.2497 ◽

2013 ◽

Vol 122 (21) ◽

pp. 2497-2497 ◽

Cited By ~ 1

Author(s):

Kui Nie ◽

Taotao Zhang ◽

Jiong Yan ◽

Leonardo Boiocchi ◽

Shuhua Cheng ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Line ◽

Cell Lines ◽

Cell Lymphoma ◽

Methylation Status ◽

B Cell Lymphoma ◽

B Cell Lymphomas ◽

Intron 1 ◽

Large B Cell

Abstract A novel IGH-associated reciprocal translocation, t(4;14)(q24;q32), was identified, along with trisomy 9, in 20 of 20 metaphases by conventional karyotyping in a case of malignant gastric post-transplant lymphoproliferative disorder (PTLD). Cloning of the translocation site by inverse PCR identified BANK1 (B-cell scaffold protein with ankyrin repeats 1), a B-cell-specific adaptor protein with putative functions in B-cell receptor and CD40 signaling, as a novel IGH translocation partner. The breakpoints were located at the Sα region of IGH and intron 1 of BANK1. The translocation juxtaposed the two genes in opposite orientations, and surprisingly, resulted in transcriptional inactivation of BANK1 as a result of dissociation of the major BANK1 promoter. While BANK1 isoforms were expressed in all tonsillar B-cells, with lower levels (∼ 5 fold) in the germinal centers (GC) compared to naïve and memory B-cells, transcription from the major promoter in the tumor was absent and transcription from the minor promoter was reduced 50% relative to GC B-cells, suggesting that the non-translocated BANK1 allele was also inactivated. The total BANK1 expression was very low (∼10% of normal GC B cells) and crytic promoter activation was not identified. Several genes (PPP3CA, MIR1255A, FLJ20021 and SLC39A8), located 180 to 440 kb away from BANK1, were analyzed for mRNA expression; there is no significant activation in any of these genes, further supporting that BANK1is indeed the target gene affected by the translocation. Interphase FISH using break-apart BANK1 probes confirmed breakpoint in the index case but did not identify translocations in additional 15 PTLDs and 68 diffuse large B-cell lymphomas (DLBCL), implying that BANK1 translocation may be a rare event. To determine if BANK1 inactivation may occur in B-cell lymphomas by other mechanisms, 23 B-cell lymphoma cell lines, including 8 Burkitt lymphoma (BL), 9 diffuse large B cell lymphoma (DLBCL), 3 primary effusion lymphoma (PEL), and 3 classical Hodgkin lymphoma (cHL) were bisulfite sequenced to assess the methylation status of 37 CpG dinucleotides in a 436 base-pair region at the 5’ end of BANK1, which extends across exon 1 into the 5’ portion of intron 1. High level of methylation (>60% methylation on average among all CpGs) was seen in all 3 cHL and 2 of 3 PEL cell lines. Regional methylation was seen in 3 of 8 BL lines and 1 of 3 PEL lines. No hypermemethylation was identified in the DLBCL lines or in normal tonsils. Hypermethylation was associated with almost complete silencing of BANK1 transcription. In the DLBCL lines and BL lines without BANK1 hypermethylation, BANK1mRNA expressions were variable, ranging from <5% to 130% of GCB cells. To confirm that BANK1 hypermethylation is present in primary lymphoma cases, methylation status of 17 of the 37 CpGs were assessed in 23 cHL cases using en bloc formalin-fixed, paraffin-embedded materials and also laser-capture micro-issected Hodgkin/Reed-Sternberg (HRS) cells. There was evidence of BANK1 hypermethylation in the tumor cells in 9 of 23 cHL. Tumor cell specificity of BANK1 hypermethylation was further confirmed in 4 cHL cases using micro-dissected HRS cells. HRS cells were negative for BANK1 in 28 of 29 cHL cases examined by immunohistochemistry, suggesting that other mechanisms other than DNA methylation may be responsible for silencing BANK1expression. To investigate whether BANK1 has biological effects on B-cells related to lymphoma development, exogenous BANK1 was re-introduced to BC3, a PEL cell line showing marked BANK1 hypermethylation with absence of BANK1 expression. We established a stable doxycycline-inducible BC3 cell line expressing BANK1. Inhibition of cell growth was observed 2 to 3 days after doxycyline induction, and the number of viable cells with transfected BANK1 was only 25% compared to BC3 cells carry vehicle alone at day 6. An analysis of 5-bromo-2’ deoxyuridine (BrdU) incorporation after 48 hours of doxycline induction revealed that the fraction of cells in S-phase was reduced by 50% in the BANK1 transfectants, suggesting that BANK1has a negative effect on cell proliferation in these B cells. In summary, we have identified a novel IGH translocation partner and provide an example of an unusual consequence (gene inactivation) of IGH-associated translocation. We provide for the first time evidence of a potential role of BANK1 down-regulation in the development of B-cell lymphomas. Disclosures: No relevant conflicts of interest to declare.

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LAG3: a novel immune checkpoint expressed by multiple lymphocyte subsets in diffuse large B-cell lymphoma

Blood Advances ◽

10.1182/bloodadvances.2019001390 ◽

2020 ◽

Vol 4 (7) ◽

pp. 1367-1377 ◽

Cited By ~ 8

Author(s):

Colm Keane ◽

Soi C. Law ◽

Clare Gould ◽

Simone Birch ◽

Muhammed B. Sabdia ◽

...

Keyword(s):

Gene Expression ◽

T Cells ◽

B Cells ◽

B Cell ◽

Cell Lymphoma ◽

International Prognostic Index ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

The Impact ◽

Large B Cell

Abstract Blockade of the PD-1 axis has modest efficacy in diffuse large B-cell lymphoma (DLBCL), but data regarding LAG3 are sparse. The impact of LAG3 digital gene expression was tested in 309 patients with DLBCL treated with standard chemoimmunotherapy. Cellular distribution of LAG3 protein was determined by immunohistochemistry and flow cytometry. In tumor-infiltrating lymphocytes (TILs), LAG3 expression was highest on CD4+ regulatory T cells (Tregs) and was also highly expressed on CD8+ T cells compared with CD4+ non-Tregs (both P = .008). LAG3high TILs were enriched in PD-1 and TIM-3. LAG3 was also expressed on a proportion of malignant B cells, and these patients had significantly higher LAG3 messenger RNA in their biopsies (P = .03). LAG3high gene expression was associated with inferior survival in discovery/validation cohorts, independent of cell of origin and the international prognostic index. Patients who were PD-L1high were fivefold more likely to be LAG3high (P < .0001). Patients who were LAG3high/PD-L1high had an inferior progression-free survival (P = .011) and overall survival (P = .005) compared with patients who were LAG3low/PD-L1high. Digital spatial protein analysis confirms LAG3 expression on T cells and, surprisingly, tumor-associated macrophages (TAMs) at higher levels than found on CD20+ B cells in the tumor microenvironment. LAG3 is frequently expressed on CD4+ Tregs and CD8+ TILs, typically with other immune checkpoints, and is also present in a proportion of malignant B cells in DLBCL and in areas enriched for TAMs. LAG3high expression is associated with poor outcome independent of conventional prognosticators.

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TCL1: a shared tumor-associated antigen for immunotherapy against B-cell lymphomas

Blood ◽

10.1182/blood-2011-09-382838 ◽

2012 ◽

Vol 120 (8) ◽

pp. 1613-1623 ◽

Cited By ~ 25

Author(s):

Jinsheng Weng ◽

Seema Rawal ◽

Fuliang Chu ◽

Hyun Jun Park ◽

Rakesh Sharma ◽

...

Keyword(s):

T Cells ◽

B Cells ◽

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Lymphocytic Leukemia ◽

Patient Specific ◽

Autologous Tumor ◽

B Cell Lymphomas ◽

B Cell Malignancies

Abstract Immunotherapy with therapeutic idiotype vaccines offers promise for treatment of B-cell malignancies. However, identification of novel immunogenic lymphoma-associated antigens that are universally expressed is necessary to overcome the barriers of patient-specific idiotype vaccines. Here, we determined whether T-cell leukemia/lymphoma 1 (TCL1) oncoprotein encoded by the TCL1 gene could be a target for immunotherapy of B-cell malignancies. We show that TCL1 mRNA and protein are selectively expressed in normal B cells but markedly hyperexpressed in multiple human B-cell lymphomas, including follicular lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma, diffuse large B-cell lymphoma, and splenic marginal zone B-cell lymphoma. We demonstrated that TCL1-specific CD8+ T cells can be generated from HLA-A*0201 (HLA-A2)+ normal donors and identified TCL171-78 (LLPIMWQL) as the minimal epitope recognized by these T cells. More importantly, TCL171-78 peptide-specific T cells were present in the peripheral blood and tumor-infiltrating lymphocytes of lymphoma patients, could be expanded in vitro, and lysed autologous tumor cells but not normal B cells in an HLA-A2–restricted manner. Our results suggest that TCL1 is naturally processed and presented on the surface of lymphoma cells for recognition by cytotoxic T cells and can serve as a novel target for development of immunotherapeutic strategies against common B-cell lymphomas.

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