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Background
Lymphomas are broadly divided into Hodgkin's Lymphoma and Non-Hodgkin's Lymphoma. They are malignant proliferations of cells which are normally found in lymphoid organs such as lymph nodes, tonsils, spleen, and thymus gland. In addition, lymphomas may arise in every other type of organ such as the brain or stomach, and these are termed extra-nodal lymphomas. One of the most exciting areas of research has focused on the role of viruses as the cause of two types of lymphoma. The HTLV-1 virus, a retrovirus similar to HIV, has been definitely implicated in the pathogenesis of the rare Adult T cell leukemia/lymphoma. The Epstein-Barr virus, the agent that causes infectious mononucleosis, has been implicated in Burkitt's lymphoma. Over the years, classification systems have evolved from purely morphologic systems combined with prognostic categories (Working Classification) to classification systems which integrate immunohistochemical data (REAL and WHO Classifications).
Cutaneous Lymphomas (CTCL, Mycosis Fungoides, Sezary Syndrome)
Enteropathy-type T-cell Lymphoma
Extranodal LymphomasHodgkin's Disease
Hodgkin's Disease-Classic Type
Hodgkin's Disease-Nodular Lymphocyte Predominance TypeAnaplastic Large Cell Lymphoma (CD30 Positive Lymphoma)
Histiocytic Lymphoma
Interdigitating Dendritic Cell Sarcoma (Reticulum Cell Sarcoma)
MALTOMA (Mucosal Associated Lymphoid Tissue Lymphomas)
Mediastinal Large B-Cell Lymphoma
Natural Killer/T-cell Lymphoma (CD56 Lymphoma)Non-Hodgkin's Lymphoma
- Burkitt Lymphoma
- Diffuse Large B-Cell Lymphoma
- Follicular Center Cell Lymphoma
- Lymphoplasmacytic Lymphoma (Immunocytoma)
- Small Lymphocytic Lymphoma
- Lymphoblastic Lymphoma
- Marginal Zone Lymphoma (Monocytoid B-Cell Lymphoma)
- Mantle Cell Lymphoma
- T cell rich B Cell Lymphoma
- Waldenstrom's Macroglobulinemia
Peripheral T-cell Lymphomas
Primary Effusion Lymphoma (Body Cavity Lymphoma)
Pyothorax-Associated Lymphoma
Splenic Marginal Zone LymphomaOUTLINE
PATHOGENESIS LYMPHOMA TYPE SPECIFIC CHROMOSOMAL TRANSLOCATION IMPLICATED ONCOGENE OR TUMOR SUPPRESSOR GENE Follicular lymphoma t(14;18)(q32;q21) bcl2 Lymphoplasmacytic lymphoma t(9;14)(p13;q32) pax5 Mantle cell lymphoma t(11;14)(q13;q32) bcl1 Extranodal marginal zone B-cell lymphoma of MALT type t(11;18)(q21;q21)
t(1:14)(p22;q32)
API2, MLT
bcl10Diffuse large B-cell lymphoma t(3;14)(q27;q32) and translocations involving 3q27 with a number of chromosome partner bcl6 Burkitt lymphoma t(8;14)(q24;q32)
t(8;22)(q24;q11)
t(2;8)(p12;q24)c-myc Myeloma t(6;14)(p25;q32) mum1 Precursor T-lymphoblastic lymphoma/leukemia t(1;14)(p32-34;q11) tal1
Anaplastic large cell lymphoma, primary systemic form t(2;5)(p23;q35)
Variant translocations involving 2p23 eg. t(1;2), t(2;3), inv(2)(p23;q35)npm, ALK
ALK fused with other partner genes such as TPM3, TFG, ATIC, CLTCL, MSN
ANGIOGENESIS
Expression of angiogenic factors and hypoxia inducible factors HIF 1, HIF 2 and CA IX in non-Hodgkin's lymphoma.Stewart M, Talks K, Leek R, Turley H, Pezzella F, Harris A, Gatter K.
Nuffield Department of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford, UK, ICRF Medical Oncology Unit, University of Oxford, John Radcliffe Hospital, Oxford, UK
Histopathology 2002 Mar;40(3):253-60 Abstract quote Expression of angiogenic factors and hypoxia inducible factors HIF 1, HIF 2 and CA IX in non-Hodgkin's lymphoma
Aims: Angiogenesis in solid tumour pathology is well established but less is known about its role in haematological malignancies. Our study investigated the immunohistochemical expression of a variety of angiogenic and hypoxic factors and microvessel densities on 110 cases of high- and low-grade non-Hodgkin's lymphomas and reactive lymphoid tissues.
Methods and results: Expression of vascular endothelial growth factor (VEGF) was present in 82 (96%) of the non-Hodgkin's cases and 35 (100%) of the reactive lymphoid tissue cases. Both hypoxia inducible factors 1alpha and 2alpha (HIF 1alpha, 2alpha) were weakly expressed in the majority of high- and low-grade lymphomas. Carbonic anhydrase IX (CA IX), a HIF-inducible membrane-bound enzyme, expression was not abundant with membranous staining being present in seven (8%) of the lymphoma cases and none of the reactive cases. Thymidine phosphorylase (TP) was distributed amongst macrophages and follicular dendritic cells but was not present in the neoplastic population. The vasculature was stained using CD34 which gave rise to a distinct vascular, predominantly paracortical network present in low-grade lymphomas and reactive lymphoid tissue but which was lost in high-grade lymphomas.
Conclusion: Our results suggest that non-Hodgkin's lymphomas may be less angiogenic and hypoxically driven than most solid tumours, which has implications for possible future therapies.
Bcl-6 Distribution and pattern of BCL-6 mutations throughout the spectrum of B-cell neoplasia.
Capello D, Vitolo U, Pasqualucci L, Quattrone S, Migliaretti G, Fassone L, Ariatti C, Vivenza D, Gloghini A, Pastore C, Lanza C, Nomdedeu J, Botto B, Freilone R, Buonaiuto D, Zagonel V, Gallo E, Palestro G, Saglio G, Dalla-Favera R, Carbone A, Gaidano G.
Divisions of Internal Medicine and Epidemiology, Department of Medical Sciences, Amedeo Avogadro University of Eastern Piedmont, Novara, Italy.
Blood 2000 Jan 15;95(2):651-9 Abstract quote
BCL-6 mutations are accumulated during B-cell transit through the germinal center (GC) and provide a histogenetic marker for B-cell tumors.
On the basis of a comprehensive analysis of 308 B-cell neoplasms, we (1) expand the spectrum of tumors associated with BCL-6 mutations; (2) corroborate the notion that mutations cluster with GC and post-GC B-cell neoplasms; and (3) identify heterogeneous mutation frequency among B-lineage diffuse large cell lymphoma (B-DLCL) subsets.
Mutations are virtually absent in acute lymphoblastic leukemia (P <.001) and mantle cell lymphoma (P <.05), whereas they occur frequently in GC or post-GC neoplasms, including lymphoplasmacytoid lymphoma, follicular lymphoma, MALT lymphomas, B-DLCL and Burkitt lymphoma. Among B-DLCL, mutations occur frequently in systemic nodal B-DLCL, primary extranodal B-DLCL, CD5(+) B-DLCL, CD30(+) B-DLCL, and primary splenic B-DLCL, suggesting a similar histogenesis of these B-DLCL subsets.
Conversely, mutations are rare in primary mediastinal B-DLCL with sclerosis (10.0%; P <.01), supporting a distinct histogenesis for this lymphoma. Longitudinal follow-up of B-DLCL transformed from follicular lymphoma shows that they BCL-6 mutations may accumulate during histologic progression. Mutations also occur in some B-cell chronic lymphocytic leukemias, small lymphocytic lymphomas, and hairy cell leukemias, consistent with the hypothesis that a fraction of these lymphoproliferations are related to GC-like cells.
Finally, the molecular pattern of 193 mutational events reinforces the hypothesis that mutations of BCL-6 and immunoglobulin genes are caused by similar mechanisms.
HER-2/NEU
Lack of HER-2/neu expression in Hodgkin and non-Hodgkin lymphoma.Bairey O, Pazgal I, Okon E, Shaklai M, Morgenshtern S.
Institutes of Hematology, Rabin Medical Center, Beilinson Campus, Petah Tiqva, Israel.
Arch Pathol Lab Med 2002 May;126(5):574-6 Abstract quote OBJECTIVE: Overexpression of HER-2/neu oncoprotein, a tyrosine kinase receptor, occurs in a variety of human cancers and has been shown to play a critical role in their development. This overexpression is usually associated with poor clinical outcome. The significance of HER-2/neu in lymphomas is unknown. The aim of this study was to evaluate the expression of HER-2/neu in the malignant lymphomas: non-Hodgkin and Hodgkin lymphomas.
METHODS: We studied formalin-fixed, paraffin-embedded tissue from 50 patients with lymphoma. Forty-two specimens were from patients with various types of non-Hodgkin lymphoma, and 8 were from patients with Hodgkin lymphoma. HER-2/neu expression was examined by an immunohistochemical technique using the HercepTest.
RESULTS: None of the specimens demonstrated overexpression or even any expression of HER-2/neu. Reactive plasma cells showed cytoplasmic staining, which was not found in malignant plasma cells from patients with multiple myeloma.
CONCLUSIONS: Non-Hodgkin and Hodgkin lymphomas do not express the HER-2/neu oncoprotein. This finding suggests that HER-2/neu does not play a role in these diseases.
OCTAMER BINDING TRANSCRIPTION FACTORS
Analysis of Octamer-Binding Transcription Factors Oct2 and Oct1 and their coactivator BOB.1/OBF.1 in Lymphomas.Saez AI, Artiga MJ, Sanchez-Beato M, Sanchez-Verde L, Garcia JF, Camacho FI, Franco R, Piris MA.
Molecular Pathology Program, Centro Nacional de Investigaciones Oncologicas Carlos III, Madrid, Spain.
Mod Pathol 2002 Mar;15(3):211-20 Abstract quote Oct1 and Oct2 are transcription factors of the POU homeo-domain family that bind to the Ig gene octamer sites, regulating B-cell-specific genes. The function of these transcription factors is dependent on the activity of B-cell-restricted coactivators such as BOB.1/OBF.1. Independent studies of the expression of these proteins in non-Hodgkin's lymphoma have been restricted to single markers, and most lack data concerning immunohistochemical expression.
Thus, we have investigated the expression of Oct1, Oct2, and BOB.1/OBF.1 in human reactive lymphoid tissue and in a series of 140 Hodgkin and non-Hodgkin's lymphomas. None of these proteins was found to be restricted to B cells, although only B cells expressed high levels of all three markers. Additionally, germinal center B cells showed stronger Oct2 and BOB.1/OBF.1 staining. Consequently, most B-cell lymphomas showed reactivity for all three antibodies. Oct2 expression was significantly higher in germinal center-derived lymphomas, although other B-cell lymphomas also displayed a high level of Oct2 expression. Although T-cell lymphomas and Hodgkin's lymphomas expressed some of these proteins, they commonly exhibited less reactivity than B-cell lymphomas.
Despite not being entirely cell-specific, the strong nuclear expression of Oct2 and BOB.1/OBF.1 by germinal center- derived lymphomas makes these antibodies a potentially useful tool in lymphoma diagnosis.
PI-3-KINASE-PTEN-AKT
Analysis of the PI-3-Kinase-PTEN-AKT pathway in human lymphoma and leukemia using a cell line microarray.Abbott RT, Tripp S, Perkins SL, Elenitoba-Johnson KS, Lim MS.
Division of Anatomic Pathology, Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, Utah 84132, USA.
Mod Pathol. 2003 Jun;16(6):607-12. Abstract quote Tissue microarray technology facilitates rapid assessment of expression of molecular markers by enabling the simultaneous analysis of hundreds of tissue specimens.
We have applied this technology to establish a microarray composed of cell pellets derived from 40 human lymphoma/leukemia-derived cell lines harboring a variety of molecular abnormalities. The application of cell line microarrays to the assessment of biologic marker evaluation was validated by studying the immunohistochemical expression of PTEN and phosphorylated AKT, two mediators of the phosphatidylinositol (PI)-3-kinase pathway. In addition to the high throughout analysis of protein expression in lymphoma/leukemia cells, this methodology also enables the evaluation of subcellular localization of protein expression.
Cytoplasmic PTEN expression was observed in the majority of cell lines (87%), whereas a minor subset demonstrated nuclear expression. Phosphorylated AKT was also expressed predominantly within the cytoplasm in 65% of cell lines, whereas nuclear expression was seen in a minority. An inverse relationship between PTEN and phosphorylated AKT was observed in 63% of cell lines. No cell lines showed absence of PTEN expression, whereas 50% of cell lines showed low PTEN expression.
Our data support the integrity of the PI-3-kinase-PTEN-AKT pathway in a majority of cell lines derived from hematologic malignancies and clearly demonstrates the utility of microarray technology in the in situ assessment of expression of molecular markers in tumor-derived cell lines.RB2/p130 TUMOR SUPPRESSOR GENE
Expression of RB2/p130 tumor-suppressor gene in AIDS-related non-Hodgkin's lymphomas: implications for disease pathogenesis.Lazzi S, Bellan C, De Falco G, Cinti C, Ferrari F, Nyongo A, Claudio PP, Tosi GM, Vatti R, Gloghini A, Carbone A, Giordano A, Leoncini L, Tosi P.
Institute of Pathological Anatomy and Histology, University of Siena, Siena, Italy.
Hum Pathol 2002 Jul;33(7):723-31 Abstract quote In this study we examined 21 cases of AIDS-related lymphomas for genomic organization and expression of RB2/p130 oncosuppressor gene and compared the results with the proliferative features of these neoplasms.
We found no mutations in the RB2/p130 gene and unusually high percentages of cells expressing nuclear pRb2/p130 in tumors with a high proliferative activity, such as AIDS-related lymphomas. These findings might suggest that a molecular mechanism usually observed in viral-linked oncogenesis could be involved. We performed in vitro and in vivo binding assays to investigate whether the human immunodeficiency virus (HIV) gene product Tat and Rb2/p130 could interact. The results of these assays revealed that the HIV-1 Tat protein binds specifically to pRb2/p130. This may result in the inactivation of its oncosuppressive properties and the induction of genes needed to proceed through the cell cycle including p107, cyclin A, and cyclin B.
Using single-cell polymerase chain reaction (PCR) assay, we found HIV-1 DNA in the neoplastic cells of only 2 of the 21 cases examined, whereas PCR on whole tissue revealed HIV-1 DNA in all of the cases. Furthermore, a diffuse and nuclear stain was observed in tissue sections with anti-Tat monoclonal antibody. These findings are in accordance with the notion that soluble Tat protein could function as a biologically active extracellular protein released by infected cells and taken up readily by uninfected B cells. In conclusion, our results seem to suggest that pRb2/p130 oncosuppressor protein may be a target in the interaction between the HIV-1 gene products and host proteins.
TCL1 Regulation of TCL1 expression in B- and T-cell lymphomas and reactive lymphoid tissues.
Narducci MG, Pescarmona E, Lazzeri C, Signoretti S, Lavinia AM, Remotti D, Scala E, Baroni CD, Stoppacciaro A, Croce CM, Russo G.
Istituto Dermopatico dell'Immacolata, Istituto di Ricovero e Cura a Carattere Scientifico, Rome, Italy.
Cancer Res 2000 Apr 15;60(8):2095-100 Abstract quote
Chromosomal rearrangements observed in T-cell prolymphocytic leukemia involve the translocation of one T-cell receptor gene to either chromosome 14q32 or Xq28, deregulating the expression of cellular protooncogenes of unknown function, such as TCL1 or its homologue, MTCP1. In the human hematopoietic system, TCL1 expression is predominantly observed in developing B lymphocytes, whereas its overexpression in T cells causes mature T-cell proliferation in transgenic mice.
In this study, using a newly generated monoclonal antibody against recombinant TCL1 protein, we extended our analysis mainly by immunohistochemistry and also by fluorescence-activated cell sorting and Western blot to a large tumor lymphoma data bank including 194 cases of lymphoproliferative disorders of B- and T-cell origin as well as reactive lymphoid tissues.
The results obtained show that in reactive lymphoid tissues, TCL1 is strongly expressed by a subset of mantle zone B lymphocytes and is expressed to a lesser extent by follicle center cells and by scattered interfollicular small lymphocytes. In B-cell neoplasia, TCL1 was expressed in the majority of the cases, including lymphoblastic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma, follicular lymphoma, Burkitt lymphoma, diffuse large B-cell lymphoma (60%), and primary cutaneous B cell lymphoma (55%). TCL1 expression was observed in both the cytoplasmic and nuclear compartments, as confirmed by Western blot analysis. Conversely, TCL1 was not expressed in Hodgkin/Reed-Sternberg cells, multiple myelomas, marginal zone B-cell lymphomas, CD30+ anaplastic large cell lymphoma, lymphoblastic T-cell lymphoma, peripheral T-cell lymphoma, and mycosis fungoides.
These data indicate that TCL1 is expressed in more differentiated B cells, under both reactive and neoplastic conditions, from antigen committed B cells and in germinal center B cells and is down-regulated in the latest stage of B-cell differentiation.
THYMUS AND ACTIVATION REGULATED CHEMOKINE
Differential Expression of Thymus and Activation Regulated Chemokine and Its Receptor CCR4 in Nodal and Cutaneous Anaplastic Large-Cell Lymphomas and Hodgkin's Disease.Vermeer MH, Dukers DF, Ten Berge RL, Bloemena E, Wu L, Vos W, De Vries E, Tensen CP, Meijer CJ, Willemze R.
Departments of Dermatology (MHV, EdV, CPT, RW) and Pathology (DFD, RLtB, EB, WV, CJLMM), Vrije Universiteit Amsterdam, the Netherlands.
Mod Pathol 2002 Aug;15(8):838-44 Abstract quote Recent studies demonstrated that Hodgkin/Reed-Sternberg (H/RS) cells in Hodgkin's disease (HD) express thymus and activation-regulated chemokine (TARC), whereas reactive lymphocytes surrounding H/RS cells express its ligand, CC-chemokine receptor 4 (CCR4). Because in vitro studies showed that CCR4 expression is a marker for lymphocytes bearing a T-helper 2 (Th2) phenotype, it was suggested that expression of TARC is a new immune escape mechanism in HD.
To find out whether this mechanism might also be operative in CD30+ malignant lymphomas other than HD, TARC and CCR4 expression was investigated by immunohistochemistry on paraffin and frozen-tissue sections of 39 nodal CD30+ anaplastic large cell lymphomas (ALCL), including 27 ALK-negative and 12 ALK-positive ALCL, 25 primary cutaneous CD30+ ALCL, including 11 patients with lymphomatoid papulosis, and 31 cases of HD. TARC was expressed by the neoplastic cells in 12/27 (44%) nodal ALK-negative ALCL and all cases of classic HD, but not in nodal ALK-positive ALCL (0/12) and only rarely in primary cutaneous CD30+ ALCL (3/25). In contrast, CCR4 was expressed by the neoplastic cells in 9/9 cutaneous CD30+ ALCL, and in 9/15 (60%) nodal ALK-negative ALCL, but only in 1/4 (25%) nodal ALK-positive ALCL and not by the H/RS cells in HD (0/8). Apart from three cases of HD showing 10 to 15% CCR4-positive lymphocytes surrounding TARC-positive H/RS cells, CCR4-positive reactive T cells were few (<5%) in all other cases studied.
Our results demonstrate a differential expression of TARC and CCR4 in different types of CD30+ malignant lymphomas. The small number of CCR4-positive reactive T cells in most cases studied argues against an important role of TARC expression in the evasion of antitumor responses.
LABORATORY/
RADIOLOGIC/OTHERCHARACTERIZATION IgH PCR Contribution of IgH-PCR to the Evaluation of B-Cell Lymphoma Involvement in Paraffin-Embedded Bone Marrow Biopsy Specimens
Richard BraunschweigMD, , Audrey Sylvia BaurMD, , Françoise DelacrétazMD, , Charlotte Bricod, and Jean BenhattarPhDAm J Clin Pathol 2003;119:634-642 Abstract quote We investigated whether the determination of clonality by polymerase chain reaction (PCR) analysis of immunoglobulin heavy chain (IgH) gene rearrangements could be helpful in the evaluation of B-cell lymphoma (BCL) involvement of bone marrow (BM) biopsy specimens.
We evaluated 83 paraffin-embedded BM biopsy specimens from 26 patients with BCL. When BM biopsy specimens considered positive, "suspicious," or negative by morphologic and immunohistochemical examination were evaluated by PCR, a monoclonal B-cell population was detected in 81% (39/48), 64% (9/14), and 11% (2/18), respectively. In most cases, a reproducible monoclonal IgH gene rearrangement was observed from BM and extramedullary sites. Nevertheless, in 4 cases, a different and independent monoclonal IgH rearrangement was observed during the disease course.PCR is efficient and complementary to morphologic and immunohistochemical examination for the evaluation of BCL involvement of BM biopsy specimens, especially when a reproducible rearrangement is found in 2 different samples.
p53 The circulating auto-antibodies to p53 protein in the follow-up of lymphoma patients.
Jezersek B, Rudolf Z, Novakovic S.
Department of Medical Oncology, Institute of Oncology, 1000 Ljubljana, Slovenia.
Oncol Rep 2001 Jan-Feb;8(1):77-81 Abstract quote
Mutant p53 proteins may be targets of the host immune system - consequently a certain proportion of cancer patients (the percentage varies according to the type of cancer) with tumors that carry p53 missense mutations develop circulating p53 antibodies.
The present study was aimed at defining the occurrence of circulating antibodies to p53 protein in patients with various types of non-Hodgkin's lymphomas (NHL). Altogether, the sera of 108 cases with various histological types of NHL and of 20 healthy controls were assessed for the presence of antibodies to p53 protein with an ELISA method. In 73 cases of NHL, also the immunohistochemical staining for p53 antigen was performed to make a rough estimation of the frequency of mutational events. The development of autoantibodies to p53 protein was observed in approximately 7% of NHL patients (predominantly in the more aggressive variants of the disease, but also in one case of small lymphocytic lymphoma). This proportion represents roughly one third of the number of patients assessed (immunohistochemically) to carry a missense p53 mutation in their tumors.
The autoantibodies to p53 protein can be used as a tumor marker (early appearance, high specificity) in the follow-up of a subset of NHL patients, but, unfortunately, this subset comprises only approximately 7% of NHL patients.
FLOW CYTOMETRY
Xian-Feng Zhao, MD, PhD, Sindhu Cherian, MD, Rachel Sargent, MD, Raja Seethala, MD, Hugh Bonner, MD, Bernard Greenberg, MD, and Adam Bagg, MD
Immunophenotypic analysis is useful in distinguishing reactive from neoplastic lymphoproliferations, particularly when tissue is limited or histologic findings are equivocal. Surface membrane immunoglobulin (SmIg) light chain restriction in B cells is especially helpful in documenting clonality, and the loss of SmIg by B cells in extramedullary sites also has been used as a criterion to support the presence of lymphoma.
However, we identified 3 cases of benign follicular hyperplasia (in 101 cases analyzed) with profound expansions (56%-88% of the B cells) of SmIg light chain–negative B cells without clonality by immunoglobulin heavy chain gene polymerase chain reaction. Thus, although uncommonly encountered, lack of SmIg light chain expression by B cells should not necessarily be interpreted as indicative of lymphoma. Interestingly, 2 of the 3 patients with these "aberrant" expansions were HIV+, and such patients are at heightened risk for the development of lymphoma.
Therefore, there is the potential for misdiagnosing lymphoma if flow cytometric data are interpreted inappropriately in isolation.
Steven J. Kussick, MD, PhD, Michael Kalnoski, MD, Rita M. Braziel, MD, and Brent L. Wood, MD, PhD
We describe 6 cases from the University of Washington Hematopathology Laboratory (Seattle) in which prominent, clonal, follicle center B-cell populations were identified by flow cytometry and confirmed by molecular methods, but in which the histologic features showed reactive follicular hyperplasia without evidence of bcl-2 overexpression or the t(14;18).
The 6 cases included 5 lymph node biopsy specimens and 1 tonsillectomy specimen. Of the 6 cases, 5 occurred in young males (8-28 years) with no known immunologic abnormality; the other case was a 32-year-old, HIV-positive woman. In all 6 cases, clonal CD10+ B cells representing at least 20% of the total B cells were identified. Available clinical follow-up ranging from 13 to 56 months revealed no evidence of lymphoma in any of the 6 patients.
Our findings add rare cases of follicular hyperplasia to the list of histologically reactive settings in which clonal B-cell populations might be present.
Comparative analysis of light chain expression in germinal center cells and mantle cells of reactive lymphoid tissues. A four-color flow cytometric study.Reichard KK, McKenna RW, Kroft SH.
Department of Pathology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75930-9072, USA.
Am J Clin Pathol 2003 Jan;119(1):130-6 Abstract quote We studied skewing of light chain ratios (LCRs) in germinal center cells (GCCs) relative to mantle cells (MCs) by flow cytometry (FC) in 98 reactive lymphoid tissues. LCRs were assessed using a 4-color lambda/kappa/CD20/CD38 tube. GCCs and MCs were discriminated by CD20 and CD38 density.
Of 98 cases, 65 contained distinct GCCs and MCs. Light chain expression usually was dimmer on GCCs than on MCs; in 22 cases, the kappa and lambda clusters converged and accurate LCRs could not be determined. Of the remaining 43 cases, the mean GCC LCR was 1.78 (range, 1.10-3.07) vs 1.56 (range, 1.00-2.24) in the MCs (P = .001). The overall kappa/lambda ratio in cases containing GCCs and MCs was 1.65 (range, 1.18-2.69) compared with 1.46 (range, 1.00-1.98) in cases containing MCs only. Of 43 cases, 19 (44%) showed differences of 20% or more between the LCRs of GCCs and MCs. LCRs of GCCs and MCs may differ substantially in reactive lymphoid tissues.
These subsets may form distinct clusters and skews in their LCRs and should not be misinterpreted as evidence of occult lymphoma.
Immunophenotypic variability of B-cell non-Hodgkin lymphoma: a retrospective study of cases analyzed by flow cytometry.Echeverri C, Fisher S, King D, Craig FE.
Department of Pathology, University of Texas Health Science Center, San Antonio, USA.
Am J Clin Pathol 2002 Apr;117(4):615-20 Abstract quote Flow cytometric analysis is important in the diagnosis, classification, and follow-up of non-Hodgkin lymphoma. It is assumed that the lymphoma phenotype for each patient remains unchanged over time and is consistent from one specimen to another. To determine the variability in expression of lymphoid antigens, we reviewed 211 flow cytometry specimens of malignant lymphoma from 81 patients. Some antigens showed a stable pattern of expression such as CD5, CD10, CD19, CD20, and HLA-DR. In contrast, CD21, CD22, CD23, and CD25 showed more variability from one specimen to another.
We believe several factors affect the stability of antigen expression. True differences in expression most probably are related to the biology and function of the different antigens. For instance, CD19 and CD20 are essential in cell maturation and function and, therefore, are present on the majority of cells. In contrast, CD22 has a role during B-cell activation and, therefore, is more variable.
Lack of standardization inflow cytometry procedures also is responsible for some variability. Instrument settings for adequate compensation and the criteria used to determine when an antigen is reported as positive are important considerations when evaluating flow cytometry histograms.
Lack of Surface Immunoglobulin Light Chain Expression by Flow Cytometric Immunophenotyping Can Help Diagnose Peripheral B-Cell Lymphoma
Shiyong Li, MD, PhD, James R. Eshleman, MD, PhD, and Michael J. Borowitz, MD, PhDAm J Clin Pathol 2002;118:229-234 Abstract quote
We determined the prevalence and significance of finding B cells without surface immunoglobulin (SIg) light chain expression. The flow cytometry database at Johns Hopkins Medical Institutions was searched for cases in which immunoglobulin light chain staining was performed to rule out a B-cell malignant neoplasm between January 1994 and February 2000.We excluded plasma cell dyscrasias, precursor B-cell acute lymphoblastic leukemia/lymphomas, and hematogones. Cases with more than 25% of B cells lacking SIg light chain expression were retrieved. Polymerase chain reaction assays for immunoglobulin heavy chain gene rearrangements were performed in SIg-negative cases with available tissue blocks.
We identified 36 cases; all represented lymphoma. Their diagnoses included diffuse large B-cell lymphoma (20), HIV-related lymphoma (5), follicular lymphoma (5), Burkitt lymphoma (2), monomorphic posttransplant lymphoproliferative disorder (1), chronic lymphocytic leukemia/small lymphocytic lymphoma (1), marginal zone B-cell lymphoma (1), and low grade B-cell lymphoma (1). Of the 17 SIg-negative cases with amplifiable DNAs, 12 (71%) showed a clonal immunoglobulin heavy chain gene rearrangement. SIg-negative B-cell lymphomas are rare.
Complete absence of SIg light chain expression in a mature B cell proliferation can be used as a surrogate marker to help diagnose peripheral B-cell lymphoma.
KARYOTYPING
James R. Cook, MD, PhD, etal
Classical cytogenetic studies have a critical role in the diagnosis of acute leukemias; however, they are much less widely used in lymphoma diagnosis.
To evaluate their utility in this latter setting, G-banded karyotyping was performed on 279 consecutive lymph node or tissue biopsy specimens with suspected lymphoma. Complete cytogenetic studies were successfully obtained in 177 cases (63.4%), including 115 (69.3%) of 166 hematolymphoid neoplasms. Success rates varied with the specific diagnosis (range, 33%-100%). The karyotypes were abnormal in 97 (84.3%) of the hematolymphoid neoplasms. In at least 3 cases of non-Hodgkin lymphoma, the findings contributed directly to the specific diagnosis made. In a much larger proportion of cases, characteristic but nonspecific findings were identified. Abnormalities of suggested prognostic importance in follicular lymphoma and in diffuse large B-cell lymphoma were identified in 14 (44%) of 32 cases and 8 (24%) of 33 cases, respectively.
Most karyotyped lymphomas will display abnormal findings including many that are not completely specific but support the diagnosis, some that provide additional prognostic information, and, infrequently, some that help establish a diagnosis that might otherwise have been missed.TISSUE MICROARRAY
Application of tissue microarray technology to the study of non-Hodgkin's and Hodgkin's lymphoma.Hedvat CV, Hegde A, Chaganti RS, Chen B, Qin J, Filippa DA, Nimer SD, Teruya-Feldstein J.
Laboratory of Cancer Genetics, Cell Biology Program, Departments of Pathology and Medicine; and the Division of Hematologic Oncology, Department of Biostatistics, Memorial Sloan Kettering Cancer Center, New York, NY.
Hum Pathol 2002 Oct;33(10):968-74 Abstract quote The immunohistochemical analysis of lymphoid neoplasms has led to refined classification schemes based on the profile of antigen expression and correlation with morphological, cytogenetic, molecular, and clinical features. Tissue microarrays (TMAs) are a powerful tool to rapidly characterize the phenotypic profile of a large number of samples.
We show that this technique can be readily applied to the study of lymphoma by examining the expression profile of a series of 193 B-cell non-Hodgkin's lymphomas (NHLs) and 29 Hodgkin's lymphomas (HLs) using immunohistochemistry and in situ hybridization (ISH). The NHL cases were studied for the expression of commonly used markers-including CD3, CD5, CD10, CD20, CD23, CD30, CD43, Bcl-2, and cyclin D1 by immunohistochemical staining of TMAs-and these results were compared with whole sections (WS) of the same cases.
We found a high degree of correlation between the results achieved with TMAs or WS (86% to 100% of cases). P53 and MIB-1 staining were studied, and the results were similar to that reported in the literature. HL cases were stained for CD20, CD30, CD15 (LeuM1), and latent membrane protein 1 expression, and ISH was performed using probes for EBER-1 and-2 transcripts. The results from HL cases on TMA sections matched exactly with those of WS.
We correlated cytogenetic results with immunohistochemical stains and morphology in cases of mantle cell lymphoma [t(11;14)(q13;q32)] and follicular lymphoma [t(14;18)(q32;q24)]. This extensive expression profile of B-cell NHLs and HL tissues discloses the ability of TMAs to rapidly screen a large series of cases and represents the first report of method validation for this technique in the study of lymphoma.
HISTOPATHOLOGY CHARACTERIZATION DISCORDANCE
Am J Surg Pathol. 2005 Dec;29(12):1549-57. Abstract quote
To evaluate the features of bone trephine biopsy involvement by non-Hodgkin lymphoma, 450 specimens were evaluated for percentage of marrow involvement, pattern of involvement, presence of germinal centers or follicular structures, and discordance with other involved sites.
A subset of 197 cases was evaluated for evidence of concurrent peripheral blood involvement. Follicular grade 1 lymphoma (30.4%) was the most common type to involve the marrow, followed by diffuse large B-cell lymphoma (16.0%), mantle cell lymphoma (9.3%), low-grade B-cell lymphoma, not otherwise specified (8.7%), lymphoplasmacytic lymphoma (8.4%), follicular grade 2 lymphoma (7.1%), and mature T- and NK-cell lymphomas (6.4%). A mixed pattern of infiltration was most common, followed by paratrabecular, nodular, diffuse, and interstitial patterns. Greater than 90% of follicular lymphomas had at least a focal paratrabecular infiltration pattern, but this pattern was also seen with other lymphoma types. Interstitial disease infiltration tended to correlate with lymphoplasmacytic lymphoma but was also not specific. The presence of germinal centers or follicular structures was associated with follicular lymphoma in 88% of cases.- Discordance between the bone marrow morphology and other tissue sites was observed in 24.9% of cases and was most often seen with follicular or diffuse large B-cell lymphoma. Peripheral blood involvement by lymphoma was observed in 29% of cases, found in all disease groups except for follicular grade 3 lymphoma.
This study highlights the frequency of different lymphoma patterns in the marrow, limitations of primary lymphoma classification on biopsy material alone, and the relative frequency of marrow discordance and peripheral blood involvement by marrow lymphoma.
Discrepancies in the immunophenotype of lymphoma cells in samples obtained simultaneously from different anatomic sites.Onciu M, Berrak SG, Medeiros LJ, Katz RL, Huh YO.
Department of Hematopathology, University of Texas, M.D. Anderson Cancer Center, Houston 77030, USA
Am J Clin Pathol 2002 Apr;117(4):644-50 Abstract quote Few studies have compared the immunophenotypic profiles of non-Hodgkin lymphoma (NHL) cells obtained simultaneously from different anatomic sites.
In the present study, we compared flow cytometry immunophenotypic results in 64 consecutive NHL cases in which aspiration or biopsy of 2 sites was performed within 30 days to assess the potential discrepancy rate. In 14 cases (22%), discordant antigen expression was identified, including 4 (36%) of 11 cases with discordant morphologic features and 10 (19%) of 53 cases with concordant morphologic features in the 2 samples. Discrepancies involved 1 antigen in 10 patients and 2 antigens in 4 patients. Antigens most frequently discrepant included CD5 (n = 4), FMC7 (n = 3), and CD20 (n = 3).
We conclude that the immunophenotype of NHL cells is generally stable, yet discrepancies can occur in a subset of patients. Differences in immunophenotype may relate to mechanisms of disease dissemination, influence of the microenvironment, or differential response to therapy.
SPECIAL STAINS/
IMMUNOPEROXIDASE/
OTHERCHARACTERIZATION Special stains Immunoperoxidase CD20+, CD5+, CD10-, CD23+, cyclin D1-, bcl6- CD20+, CD5+, CD10-, CD23-, cyclin D1+, bcl6- CD20+, CD5-, CD10-, cyclin D1-, bcl-6- CD20+, CD5-, CD10+, bcl6+, bcl2+, cyclin D1- CD20+, CD5-, CD10-, cIg+ CD20-, CD45RB-, cIg+, CD79a+ CD20+, CD5-/+, CD10-/+, bcl-6+/-, cIg-/+ CD20+, CD5-, CD10+, bcl-2+, MIB1++ CD20-/+, CD79a+, CD10+/-, TdT+ GENERAL
B-cell lymphomas with coexpression of CD5 and CD10.Dong HY, Gorczyca W, Liu Z, Tsang P, Wu CD, Cohen P, Weisberger J.
IMPATH Inc, 521W 57th St, 6th Floor, New York, NY 10019, USA.
Am J Clin Pathol 2003 Feb;119(2):218-30 Abstract quote Coexpression of CD5 and CD10 is highly unusual in B-cell lymphomas and may pose a diagnostic challenge.
We report 42 cases of B-cell lymphoma with simultaneous expression of CD5 and CD10. They made up approximately 0.4% of all B-cell lymphomas seen during the study period and included the following cases: large B-cell lymphoma (LBCL), 14 (33%); follicular lymphoma (FL), 10 (24%); mantle cell lymphoma (MCL), 9 (21%); chronic lymphocytic leukemia, 4 (10%); acute precursor B-cell lymphoblastic leukemia/lymphoma, 2 (5%); and other low-grade B-cell lymphomas, 3 (7%).
All MCLs had overexpression of bcl-1 or the t(11;14) and were CD43+. All FLs had typical histomorphologic features and were bcl-2+ and bcl-6+ but CD43-. Of 14 LBCLs, 5 were histologically high-grade. Six (43%) of 14 patients with LBCL died within 10 months of diagnosis of CD5+CD10+ lymphoma (median survival, 4 months), including all 3 patients with stage IV disease and 2 of 5 with histologically high-grade lymphoma. Our findings indicate that coexpression of CD5 and CD10 is rare but occurs in diverse subtypes of B-cell lymphoma.
Investigation of bcl-1, bcl-6, and CD43 and morphologic evaluation may resolve the potential confusion in diagnosis and lead to the recognition of the correct lymphoma subtype.
CD10 and BCL-6 Expression in Paraffin Sections of Normal Lymphoid Tissue and B-Cell Lymphomas Ahmet Dogan, M.D., Ph.D., MRCPath; Eniko Bagdi, M.D.; Philippa Munson, M.Sc.; Peter G. Isaacson, M.B., Ch.B., D.M., FRCPath, D.Sc.
From the Department of Histopathology, Royal Free and University College Medical School, London, U.K.
Am J Surg Pathol 2000;24:846-852 Abstract quote In this study the authors explored the value of immunostaining for follicular center B-cell markers, BCL-6 and CD10, in paraffin sections as a tool for the differential diagnosis of B-cell lymphomas.
The cases studied comprised reactive lymphoid hyperplasia (RLH; n = 19), follicular lymphoma (FL; n = 50), low-grade mucosa-associated lymphoid tissue (MALT) lymphoma (n = 24), mantle cell lymphoma (n = 19), splenic marginal zone lymphoma (n = 13), diffuse large B-cell lymphoma (DLBCL; n = 54), Burkitt's lymphoma (BL; n = 20), nodular lymphocyte predominance Hodgkin's disease (NLPHD; n = 16), and classic Hodgkin's disease (CHD; n = 13).
In RLH, CD10 and BCL-6 were expressed almost exclusively by the follicular center cells. In contrast in FL, the expression of CD10 (39/50) and BCL-6 (34/36) was seen in both follicular and interfollicular neoplastic B cells. Marginal zone/MALT lymphomas and mantle cell lymphoma were always negative. In DLBCL the expression was variable for both CD10 (21/54) and BCL-6 (39/47), with some tumors, including cases of transformed follicular lymphoma (9/10), coexpressing CD10 and BCL-6, and others expressing only BCL-6, and a small group expressing neither marker, possibly reflecting the underlying primary pathogenetic events such as the rearrangement of BCL-2 or BCL-6 genes. BL was always both CD10 and BCL-6 positive. In NLPHD the L&H cells expressed BCL-6 (11/13) but not CD10, whereas in CHD BCL-6 expression was seen in half of the cases.
This study demonstrates that both CD10 and BCL-6 are reliable markers of follicular center B-cell differentiation. CD10 and BCL-6 immunostaining have an important role in differential diagnosis of FL from RLH and other low-grade B-cell lymphomas. The results also suggest that a CD10/BCL-6 expression pattern may be helpful in identifying main subsets of DLBCL. However, additional studies comparing genotype with immunophenotype are required.
MUM1: a step ahead toward the understanding of lymphoma histogenesis.
Gaidano G, Carbone A.
Leukemia 2000 Apr;14(4):563-6 Abstract quote
In recent times, the field of B cell lymphoma histogenesis has progressed rapidly due to the increasing availability of histogenetic markers. Genotypic markers of B cell histogenesis are represented by mutations of IgV and BCL-6 genes, which are somatically acquired at the time of B cell transit through the germinal center (GC). Phenotypic markers are represented by BCL-6 and CD138/syndecan-1 protein expression and allow the distinction between GC and post-GC B cells.
On this basis, lymphomas may be histogenetically distinguished into: (1) lymphomas devoid of somatic IgV and BCL-6 hypermutation, which derive from pre-germinal center B cells; (2) lymphomas associated with somatic IgV and/or BCL-6 hypermutation and BCL-6 expression, which closely reflect germinal center B cells; and (3) lymphomas associated with somatic IgV and/or BCL-6 hypermutation, as well as CD138/syndecan-1 positivity, representing lymphomas of post-germinal center B cells.
In the March issue of Leukemia, Tsuboi et al report on the expression pattern of MUM1 in normal lymphoid tissues and in lymphoma. Because expression of MUM1 protein appears to be strictly regulated during lymphoid differentiation, and because expression of the molecule is retained upon neoplastic transformation, MUM1 may be added to the panel of phenotypic markers of B cell lymphoma histogenesis.
In particular, MUM1 may provide a marker for the identification of transition from BCL-6 positivity (GC B cells) to CD138 expression (immunoblasts and plasma cells). These studies are of potential clinical value, since in some B cell malignancies, histogenesis may influence prognosis.
Analysis of MUM1/IRF4 protein expression using tissue microarrays and immunohistochemistry.
Natkunam Y, Warnke RA, Montgomery K, Falini B, van De Rijn M.
Department of Pathology, Stanford University Medical Center, Stanford, California 94305, USA.
Mod Pathol 2001 Jul;14(7):686-94 Abstract quote
The gene encoding MUM1 was characterized as a possible translocation partner in chromosomal abnormalities involving a significant number of multiple myelomas. The overexpression of the MUM1 protein as a result of translocation t(6;14) (p25;q32) identified MUM1 as a putative regulatory molecule involved in B-cell differentiation and tumorigenesis. The expression of MUM1 protein in multiple myelomas supports this hypothesis.
In the current study, using tissue microarray technology, we have tested the expression of the MUM1 protein in 1335 human malignancies and normal tissues.
Our data show that the MUM1 protein is expressed in a wide spectrum of hematolymphoid neoplasms and in malignant melanomas but is absent in other human tumors. In addition, in tissue microarrays as well as in conventional paraffin sections, MUM1 staining was found to lack specificity in detecting plasmacytic differentiation as compared with two markers, CD138/Syndecan and VS38, commonly used in paraffin immunohistochemistry for detection of plasma cells.
Expression profile of MUM1/IRF4, BCL-6, and CD138/syndecan-1 defines novel histogenetic subsets of human immunodeficiency virus-related lymphomas.
Carbone A, Gloghini A, Larocca LM, Capello D, Pierconti F, Canzonieri V, Tirelli U, Dalla-Favera R, Gaidano G.
Divisions of Pathology and Medical Oncology A, Centro di Riferimento Oncologico, Istituto Nazionale Tumori, IRCCS, Aviano, Italy.
Blood 2001 Feb 1;97(3):744-51 Abstract quote
This study was aimed at defining the histogenesis of the pathologic spectrum of lymphoma arising in the context of human immunodeficiency virus (HIV) infection.
Toward this aim, 87 AIDS-related non-Hodgkin lymphomas (AIDS-NHL) and 16 Hodgkin lymphomas arising in HIV+ patients (HIV-HL) were comparatively analyzed for the expression pattern of several B-cell histogenetic markers, including BCL-6 (expressed by centroblasts and centrocytes),
MUM1/IRF4 (expressed by late centrocytes and post-germinal center [GC] B cells), and CD138/syn-1 (expressed by post-GC B cells). Expression of MUM1, BCL-6, and syn-1 segregated 3 major phenotypic patterns among AIDS-NHL and HIV-HL: (1) the BCL-6+/MUM1-/syn-1- pattern, selectively clustering with a large fraction of AIDS-Burkitt lymphoma (17 of 19) and of systemic AIDS-diffuse large cell lymphoma (12 of 16); (2) the BCL-6-/MUM1+/syn-1- pattern, associated with a fraction of AIDS-immunoblastic lymphoma (8 of 24); and (3) the BCL-6-/MUM1+/syn-1+ pattern, associated with systemic and primary central nervous system immunoblastic lymphoma (14 of 24) and with primary effusion lymphoma (10 of 10), plasmablastic lymphoma of the oral cavity (7 of 7), and HIV-HL (15 of 16).
Analysis of nonneoplastic lymph nodes showed that the 3 phenotypic patterns detected in AIDS-NHL and HIV-HL correspond to distinct stages of physiologic B-cell development-centroblasts (BCL-6+/MUM1-/syn-1-), late GC/early post-GC B cells (BCL-6-/MUM1+/syn-1-), and post-GC B cells (BCL-6-/MUM1+/syn-1+). Expression of the Epstein-Barr virus-encoded latent membrane protein-1 clustered with the BCL-6-/MUM1+/syn-1+ profile throughout the clinicopathologic spectrum of AIDS-NHL and HIV-HL.
Overall, these results define novel histogenetic subsets of AIDS-NHL and HIV-HL and may provide novel tools for refining the diagnosis of these disorders.
Diagnostic usefulness of CD23 and FMC-7 antigen expression patterns in B-cell lymphoma classification.
Garcia DP, Rooney MT, Ahmad E, Davis BH.
Departments of Clinical, William Beaumont Hospital, Royal Oak, MI, USA.
Am J Clin Pathol 2001 Feb;115(2):258-65 Abstract quote
CD23 and FMC-7 are normal B-cell antigens used during diagnostic immunophenotyping of suspected lymphoproliferative disorders, but the diagnostic usefulness of antigenic expression patterns of simultaneous 2-color staining and flow cytometric analysis has not been reported.
We evaluated the FMC-7 and CD23 expression pattern in 201 cases of B-cell lymphoma from tissue biopsy specimens by multiparameter flow cytometry. The CD23-/FMC-7+ pattern was the most common pattern in large cell, mantle cell, and marginal zone lymphomas. The CD23 and FMC-7 antigen, along with the CD5 coexpression pattern, permitted accurate classification of all 71 cases of small lymphocytic, mantle cell, and marginal zone types of lymphoma. The widest variation of patterns was with follicular cell lymphoma, although most cases expressed the CD23 +/-/FMC-7+ pattern (+/-, partial or minor subset expression). The CD23 and FMC-7 antigen expression pattern was predictive of subtypes in more than 95% of lymphoma cases and could narrow the differential diagnosis in the remaining cases.
We conclude the flow cytometric CD23/FMC-7 expression pattern achieved by dual staining facilitates accurate and reproducible classification of B-cell lymphomas and has diagnostic usefulness.
Comparison of Immunophenotypes of Small B-Cell Neoplasms in Primary Lymph Node and Concurrent Blood or Marrow Samples
Yin Xu, MD, PhD, Robert W. McKenna, MD, Sheryl L. Asplund, MD, and Steven H. Kroft, MDAm J Clin Pathol 2002;118:758-764 Abstract quote
Immunophenotyping of small B-cell neoplasms (SBCNs) may have a critical role in diagnosis. However, there are few data addressing whether the immunophenotypes of SBCNs in bone marrow (BM) and peripheral blood (PB) are representative of those in other tissue sites.
We compared the immunophenotypic features of concurrently analyzed lymph node (LN) and BM/PB specimens using multiparameter flow cytometry. Fifty-five SBCNs were identified: 27 follicular lymphomas (FLs), 16 chronic lymphocytic leukemia/small lymphocytic lymphomas (CLL/SLLs), and 12 mantle cell lymphomas (MCLs). Major (presence vs absence) or minor (alteration of intensity) variations in expression of individual antigens between LN and BM/PB were observed in up to 25% of cases within a particular SBCN category.
All FLs and CLL/SLLs maintained characteristic immunopheno-types in BM/PB. Potentially misleading variations included 1 case of MCL that failed to express CD5 in BM and likely would have been immunophenotypically misclassified as a marginal zone lymphoma and another MCL that expressed moderate CD23 in PB and would have required additional studies for precise classification. The remaining major and minor variations would not have affected interpretation.CCR6
CCR6 is a functional chemokine receptor that serves to identify select B-cell non-hodgkin's lymphomas.Rodig SJ, Jones D, Shahsafaei A, Dorfman DM.
Departments of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA; and the Department of Pathology, University of Texas-MD Anderson Cancer Center, Houston, TX.
Hum Pathol 2002 Dec;33(12):1227-33 Abstract quote Several recent studies have revealed important contributions of chemokines and their receptors to the development and progression of both hematopoietic and nonhematopoietic neoplasms.
The chemokine receptor CCR6 is unusual in that it mediates leukocyte chemotaxis in response to a single chemokine, CCL20 (macrophage inhibitory factor-3alpha), as well as in response to a family of antimicrobial peptides termed "beta-defensins." CCR6 is critical for mucosal immunity, and expression of the receptor is tightly regulated on hematopoietic cells.
Here we characterize the expression of CCR6 on B cells and B-cell non-Hodgkin's lymphomas. We demonstrate that CCR6 expression is limited to cells comprising the mantle and marginal zones of the secondary lymphoid tissues and serves to identify the majority of mantle cell, marginal zone, and mucosa-associated lymphoid tissue lymphomas. Furthermore, we show that CCR6 serves as a functional chemokine receptor when expressed by neoplastic cells. Finally, we establish that the cognate ligand for CCR6 is present on mucosal epithelium infiltrated by neoplastic cells in select extranodal lymphomas.
Thus, CCR6 is a useful new marker identifying a subset of B-cell non-Hodgkin's lymphomas and likely contributes to the localization of select extranodal lymphomas at mucosal sites.
CLUSTERIN
Clusterin expression in malignant lymphomas: a survey of 266 cases.Saffer H, Wahed A, Rassidakis GZ, Medeiros LJ.
Department of Pathology (HS, AW), The University of Texas-Houston Health Sciences Center.
Mod Pathol 2002 Nov;15(11):1221-6 Abstract quote Clusterin expression has been reported to be characteristic of systemic anaplastic large cell lymphoma and usually negative in cutaneous anaplastic large cell lymphoma as well as other lymphoma types.
We surveyed clusterin expression using immunohistochemical methods in 266 cases of non-Hodgkin's lymphoma and Hodgkin's disease to further assess the diagnostic utility of this marker. Clusterin immunostaining was observed in 40 of 49 (82%) systemic anaplastic large cell lymphomas and 12 of 29 (41%) cutaneous anaplastic large cell lymphomas. Clusterin also was expressed in 5 of 43 (12%) diffuse large B-cell lymphomas (4 of 5 CD30+), 1 of 14 (7%) peripheral T-cell lymphomas, 1 of 32 (3%) cases of nodular sclerosis Hodgkin's disease, and 1 case of mycosis fungoides in large cell transformation.
Clusterin was negative in all other neoplasms assessed including follicular lymphoma of all grades (n = 24), mantle cell lymphoma (n = 13), marginal zone B-cell lymphoma (n = 12), precursor T-cell or B-cell lymphoblastic leukemia/lymphoma (n = 10), mixed cellularity Hodgkin's disease (n = 8), chronic lymphocytic leukemia/small lymphocytic lymphoma (n = 7), Burkitt lymphoma (n = 7), mycosis fungoides (n = 4), nodular lymphocyte predominant Hodgkin's disease (n = 3), lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (n = 2), and plasmacytoma (n = 2).
We conclude that clusterin is a marker of anaplastic large cell lymphoma and that addition of clusterin to antibody panels designed to distinguish systemic anaplastic large cell lymphoma from classical Hodgkin's disease is useful. However, clusterin is also positive in a substantial subset of cutaneous anaplastic large cell lymphomas, a smaller subset of diffuse large B-cell lymphomas, and rarely in cases of peripheral T-cell lymphoma and nodular sclerosis Hodgkin's disease.
HEAT SHOCK PROTEIN-90
- Expression of heat-shock protein-90 in non-Hodgkin's lymphomas.
Valbuena JR, Rassidakis GZ, Lin P, Atwell C, Georgakis GV, Younes A, Jones D, Medeiros LJ.
1Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
Mod Pathol. 2005 Oct;18(10):1343-9. Abstract quote
Heat-shock protein-90 (HSP90) inhibitors are currently being used in phase I clinical trials for treating patients with a variety of neoplasms including lymphomas.
Using immunohistochemical methods, we assessed for HSP90 expression in 412 cases of non-Hodgkin's lymphoma. In B-cell lymphomas, HSP90 was moderately to strongly expressed in all cases of Burkitt's lymphoma (5/5, 100%), and in subsets of follicular lymphoma (17/28, 61%), diffuse large B-cell lymphoma (27/46, 59%), nodal marginal zone B-cell lymphoma (6/16, 38%), plasma cell neoplasms (14/39, 36%), small lymphocytic lymphoma/chronic lymphocytic leukemia (3/9, 33%), mantle cell lymphoma (12/38, 32%) and lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (3/10, 30%). HSP90 was weakly expressed in six of 14 (43%) cases of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue. In T-cell lymphomas, HSP90 was moderately to strongly expressed in subsets of anaplastic large-cell lymphoma (14/24, 58%; 9/12 ALK+ and 5/12 ALK-), precursor-T-cell lymphoblastic leukemia/lymphoma (20/65, 31%), unspecified peripheral T-cell lymphoma (8/43, 23%) and angioimmunoblastic T-cell lymphoma (2/17, 12%). HSP90 was weakly expressed in seven of 58 (12%) cases of mycosis fungoides.
We conclude that HSP90 is commonly expressed in a subset of many types of B- and T-cell lymphoma. These data suggest that many lymphoma types are suitable targets for modulation of HSP90 activity, and that HSP90 inhibitors are a potential investigational therapy for lymphoma patients.MCL-1 MCL-1 expression in B-cell non-Hodgkin's lymphomas.
Cho-Vega JH, Rassidakis GZ, Admirand JH, Oyarzo M, Ramalingam P, Paraguya A, McDonnell TJ, Amin HM, Medeiros LJ.
Department of Pathology, University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA.
Hum Pathol. 2004 Sep;35(9):1095-100. Abstract quote
B-cell non-Hodgkin's lymphomas are known to express BCL-2 family proteins, of which the myeloid cell leukemia-1 (MCL-1) protein is a member. MCL-1 is involved in viability and immortalization of normal and neoplastic B cells, and expression is regulated transcriptionally and posttranscriptionally, resulting in an anti-apoptotic (full length) or a pro-apoptotic (short isoform) gene product.
In this study, we assessed 151 B-cell lymphomas for MCL-1 expression and analyzed for expression of the full-length and short isoforms of MCL-1 in B-cell lymphoma cell lines. By using immunohistochemistry, a subset of neoplasms in 9 lymphoma types studied expressed MCL-1, but expression was more frequent and intense in high-grade (43 of 49, 88%) compared with low-grade (34 of 92, 37%) lymphomas (P < 0.0001).
In follicular lymphomas, MCL-1 expression positively correlated with increasing grade; 1 (14%) of eight grade 1, 7 (70%) of ten grade 2, and all 9 (100%) grade 3 were positive (P < 0.0008). All plasma cell myeloma cases assessed were also MCL-1 positive. By using Western blot analysis, 6 of 7 high-grade B-cell lymphoma cell lines showed predominant expression of full-length MCL-1, compared with no or weak expression of the short isoform. One myeloma and 1 of 2 mantle cell lymphoma cell lines also tested showed only full-length isoform expression.
Our data suggest that MCL-1 is frequently expressed in high-grade B-cell lymphomas and plasma cell myeloma, most likely in its full-length isoform that is an active anti-apoptotic gene product. MCL-1 expression also correlates with grade and may contribute to transformation in follicular lymphomas.PAX-5
The value of anti-pax-5 immunostaining in routinely fixed and paraffin-embedded sections: a novel pan pre-B and B-cell marker.Torlakovic E, Torlakovic G, Nguyen PL, Brunning RD, Delabie J.
Am J Surg Pathol 2002 Oct;26(10):1343-50 Abstract quote Whereas L26 (anti-CD20) is well established as a B-cell marker of high specificity for use in paraffin-embedded tissues and JCB117 (anti-CD79a) is increasingly used, a comparable additional pan-B-cell antibody has hitherto not yet been identified.
Here we have studied the use of a novel anti-pan-B-cell marker Pax-5 for use in diagnostic pathology. Pax-5 encodes for BSAP (Pax-5), a B-cell-specific transcription factor, the expression of which is detectable as early as the pro-B-cell stage and subsequently in all further stages of B-cell development until the plasma cell stage where it is downregulated. Pax-5 is essential for B-lineage commitment in the fetal liver, whereas in adult bone marrow this transcription factor is required for progression of B-cell development beyond the early pro-B (pre-BI) cell stage. Among the B-cell genes that are present in early B-cell development and are upregulated by Pax-5 are CD19 and Igalpha (CD79a).
We have tested a commercially available anti-Pax-5 antibody (anti-BSAP, clone 24) in a series of 592 routinely fixed and paraffin wax-embedded biopsies, including lymph nodes, bone marrow, and various other organs containing lymphoid tissues. Pax-5 protein (BSAP) was detected in all cases of precursor and mature B-cell non-Hodgkin lymphomas/leukemias. In addition, in 97% of classic Hodgkin lymphomas, Reed-Sternberg cells expressed Pax-5. However, Pax-5 was not detected in any of the multiple myelomas, solitary plasmacytomas, and 4% of diffuse large B-cell lymphomas. Among those diffuse large B-cell lymphomas not expressing Pax-5 were only those with terminal B-cell differentiation. All T-cell non-Hodgkin lymphomas, including ALCL and lymphoblastic lymphomas and leukemias, were negative. There was a strong association between Pax-5 and CD20 expression.
We conclude that anti-Pax-5 is an excellent pan-B and pan-pre-B-cell marker. We have found that anti-Pax-5 is superior to anti-CD20 in the diagnosis of pre-B acute lymphoblastic leukemia and classic Hodgkin lymphoma versus ALCL of T and "null" cell type. It was also useful in differential diagnosis between lymphoplasmacytic lymphoma and plasmacytoma. Even though there is an excellent correlation between CD20 and Pax-5 expression, anti-Pax-5 exceeds the specificity and sensitivity of L26 (anti-CD20) because of its earlier expression in B-cell differentiation and its ability to detect all committed B cells, including classic Hodgkin lymphoma.
TELOMERASE ACTIVITY
Telomerase activity and proliferation index in aggressive mature B-cell lymphoma: comparison to germinal center phenotypic markers.
Chiu KC, Fine M, Ikle D, Slovak ML, Arber DA.
Division of Pathology, City of Hope National Medical Center, Duarte, CA, USA.
Hum Pathol. 2003 Dec;34(12):1259-64. Abstract quote
Cell proliferation may be evaluated by various methods, including Ki-67 immunohistochemistry and measures of telomerase activity. Both methods would theoretically show comparable increases in a given case. To evaluate the relationship between these 2 markers of proliferation in aggressive mature B-cell lymphomas, 48 cases were studied.
The study group included 5 cases of mantle cell lymphoma (MCL); 6 cases of Burkitt's/Burkitt's-like lymphoma (BL); 9 cases of follicular lymphoma, grade 3 (FLC); and 28 cases of diffuse large B-cell lymphoma (DLC). Telomerase activity was measured as total product generated (TPG) units, and TPG results for the aforementioned cases were compared to the TPG results for 10 cases of reactive follicular hyperplasia. An overlap in TPG scores between reactive cases and lymphoma cases was found. Significant differences in both log TPG (P = 0.0443) and Ki-67 (P = 0.0006) were seen in the different lymphoma types. A positive correlation between Ki-67 percentage and TPG score was identified in FLC (r = 0.9281; P = 0.0003), but a poor correlation between these 2 indicators was seen in the other lymphoma types. Cluster analysis identified distinct patterns for MCL, FLC, and BL, but heterogeneous patterns for DLC. Because increases in both Ki-67 proliferation and telomerase activity are reported in normal germinal centers (GCs), these tests were also evaluated for usefulness as markers of a GC cell phenotype. Among the FLC and DLC cases, features of a GC phenotype significantly correlated with increased Ki-67 percentage (P = 0.0152), but not with increased log TPG. An elevated log TPG correlated with CD10 expression, and elevated Ki-67 percentage correlated with both CD10 and BCL-6 expression. TPG level and Ki-67 percentage did not correlate with the presence of t(14;18) or BCL-2 protein expression. Although the proliferation patterns were fairly distinctive for MCL, FLC, and BL, these studies show that markers of cell proliferation do not by themselves,identify distinct subtypes of large cell lymphomas.With the exception of FLC, the tumors exhibited poor correlation between telomerase activity and Ki-67 proliferation index. These tests did show some correlation with expression of GC cell phenotypic markers, however.
DIFFERENTIAL DIAGNOSIS KEY DIFFERENTIAL FEATURES FEATURES FAVORING LYMPHOMA OVER REACTIVE LYMPHOID HYPERPLASIAS Taken from Chan JKC. Practical Lymphoma Diagnosis: A Simplified Approach. Presented at the 111th Semi-Annual California Tumor Tissue Registry. December 2001 Effaced normal architecture
Abnormal architecture of the follicles, marginal zones, or interfollicular zones
Excessive numbers of B cells forming diffuse sheetsExtensive perinodal involvement
Interfollicular invasion (morphologically or by immunohistochemistry)
Destruction of mantles or germinal centers of reactive follicles
Epithelial destruction (lymphoepithelial lesions)
Vascular invasionAtypical cells morphologically deviating from reactive cells
Abnormal cytoplasmic or nuclear inclusions
Abnormal/aberrant immunophenotypeDDX BY PATTERN ANALYSIS Nodular lymphocyte predominance Hodgkin lymphoma
Lymphocyte-rich classical Hodgkin lymphoma, nodular
Progressive transformation of germinal centers
Follicular lymphoma, floral variantReactive follicular hyperplasia
Follicular lymphoma
Follicular/nodular form of mantle cell lymphoma
Marginal zone B cell lymphoma with follicular colonization
Castleman diseaseB-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CLL/SLL)
Lymphoplasmacytic lymphoma
Mantle cell lymphoma
Extranodal marginal zone B-cell lymphoma of MALT type
Follicular center cell lymphoma, diffuse
Splenic marginal zone B-cell lymphomaBurkitt lymphoma
Lymphoblastic lymphoma
Granulocytic sarcoma
Blastoid variant of mantle cell lymphoma
Blastic NK cell lymphoma
Blastic transformation of follicular lymphoma
Peripheral T-cell lymphoma, monomorphic medium-sized cell type
NK/T cell lymphoma (examples predominated by medium-sized cells)
Nodal marginal zone B-cell lymphoma (some cases)
Systemic mastocytosisIntravascular Large B-cell lymphoma
Plasmacytoma
Infectious mononucleosis
Enteropathy-type T-cell lymphoma
Nasal and extra-nasal NK/T cell lymphomas
Anaplastic large cell lymphoma
Lymphomatoid granulomatosis
|Sclerosing extramedullary hematopoietic tumorTrue histiocytic lymphoma
Rosai-Dorfman disease
Follicular dendritic cell tumor/sarcoma
Inflammatory pseudotumor
Inflammatory pseudotumor-like follicular dendritic cell tumor
PROGNOSIS AND TREATMENT CHARACTERIZATION PROGNOSIS Taken from Chan JKC. Practical Lymphoma Diagnosis: A Simplified Approach. Presented at the 111th Semi-Annual California Tumor Tissue Registry. December 2001. Follicular lymphoma
B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma
Lymphoplasmacytic lymphoma
Mantle cell lymphoma (worst outcome of the group)
Splenic marginal zone B-cell lymphoma
Nodal marginal zone B-cell lymphoma
Mycosis fungoides
T-cell granular lymphocytic leukemiaDiffuse large B-cell lymphoma
Various peripheral T-cell lymphomas and NK cell lymphomas except mycosis fungoides (behavior more like indolent lymphoma), T-cell granular lymphocytic leukemia (behavior more like indolent lymphoma), and primary cutaneous anaplastic large cell lymphoma (special group of localized indolent lymphoma)Burkitt lymphoma
Lymphoblastic lymphomaExtranodal marginal zone B-cell lymphoma of MALT type
Primary cutaneous anaplastic large cell lymphomaINTERNATIONAL PROGNOSTIC INDEX (IPI) FOR PROGNOSTICATION OF NON-HODGKIN'S LYMPHOMA PARAMETERS Age >/= 60 years
Advanced stage (III or IV)
>1 extranodal sites of involvement
Performance status >/=2
Serum lactate dehydrogenase level raisedRISK GROUP STRATIFICATION ACCORDING TO THE IPI (TOTAL NUMBER OF ABOVE LISTED FEATURES) Low risk Low intermediate risk High intermediate risk High risk TREATMENT Feasibility and pharmacokinetic study of a chimeric anti-CD20 monoclonal antibody (IDEC-C2B8, rituximab) in relapsed B-cell lymphoma. The IDEC-C2B8 Study Group.
Tobinai K, Kobayashi Y, Narabayashi M, Ogura M, Kagami Y, Morishima Y, Ohtsu T, Igarashi T, Sasaki Y, Kinoshita T, Murate T.
Department of Medical Oncology, National Cancer Center Hospital, Tokyo.
Ann Oncol 1998 May;9(5):527-34 Abstract quote
BACKGROUND: In clinical trials in the USA, IDEC-C2B8 (a mouse-human chimeric anti-CD20 monoclonal antibody) has demonstrated high response rates with only mild toxic effects in relapsed B-cell lymphoma at a dose of four weekly 375 mg/m2 infusions. The aim of the present trial was to determine whether or not this dose is practically applicable to Japanese patients with relapsed B-cell lymphoma with respect to safety, pharmacokinetics and efficacy.
PATIENTS AND METHODS: Patients with relapsed CD20+ B-cell lymphoma received intravenous infusions of IDEC-C2B8 once a week for four weeks. A total of 12 patients (four at 250 mg/m2 and eight at 375 mg/m2) were enrolled.
RESULTS: All 11 eligible patients treated with either dose level tolerated IDEC-C2B8 well. Commonly observed adverse drug reactions were grades 1 or 2 non-hematologic toxicities during the infusion, consisting mostly of flu-like symptoms and skin reactions. All of the observed hematologic toxicities were of grade 3 or less, and transient. A rapid and sustained B-cell decrease in peripheral blood was observed, but no infectious episodes were encountered. Human anti-mouse and anti-chimeric antibodies were not detected. Of the 11 eligible patients (eight with follicular, two with diffuse large-cell and one with mantle cell lymphoma), two showed a complete response and five showed a partial response, and all of the seven responders had lymphoma with follicular histology. A pharmacokinetic analysis showed that the elimination half-life (T1/2) of IDEC-C2B8 was 445 +/- 361 hours, and that the serum antibody levels increased in parallel with the course of infusions, and in most patients was still measurable at three months.
CONCLUSIONS: The dose of four weekly 375 mg/m2 infusions of IDEC-C2B8 is safe and effective in Japanese patients with relapsed B-cell lymphoma. Further studies evaluating IDEC-C2B8 are warranted.
High dose therapy and autologous stem cell transplantation for human immunodeficiency virus-associated non-Hodgkin lymphoma in the era of highly active antiretroviral therapy.
Molina A, Krishnan AY, Nademanee A, Zabner R, Sniecinski I, Zaia J, Forman SJ.
Division of Hematology and Bone Marrow Transplantation, City of Hope National Medical Center, Duarte, California 91010, USA.
Cancer 2000 Aug 1;89(3):680-9 Abstract quote
BACKGROUND: The advent of highly active antiretroviral therapy (HAART) has allowed the exploration of more dose-intensive therapy such as autologous stem cell transplantation (ASCT) in selected patients with human immunodeficiency virus (HIV)-associated non-Hodgkin lymphoma (NHL).
METHODS: The authors report on the use of myeloablative chemotherapy with ASCT in two HIV positive patients with NHL. The first patient underwent ASCT at the time of first disease remission for poor risk, diffuse, large cell NHL and the second patient had multiply recurrent, chemosensitive Burkitt lymphoma. ASCT was performed in both patients using a transplant conditioning regimen of high dose cyclophosphamide, carmustine, and etoposide (CBV).
RESULTS: The target dose of >/= 5 x 10(6)/kg CD34 positive peripheral blood stem cells (PBSC) utilized for ASCT was collected using granulocyte-colony stimulating factor (G-CSF) after chemotherapy for mobilization while both patients were receiving concomitant HAART for HIV infection. HAART was continued during CBV conditioning. Prompt hematopoietic recovery was observed after ASCT. Both patients remained in clinical disease remission from their lymphoma at 28 months and 20 months after transplant, respectively.
CONCLUSIONS: ASCT is feasible in patients with HIV-associated NHL. Adequate numbers of CD34 positive PBSC can be procured from patients receiving HAART and chemotherapy for NHL. Selected patients with HIV-related lymphoma can tolerate the high dose CBV myeloablative chemotherapy regimen without increased acute regimen-related toxicity. Reinfusion of G-CSF-mobilized PBSC can lead to rapid recovery of hematologic function and sustained engraftment after ASCT. Given the poor prognosis of patients with HIV-associated NHL treated with conventional chemotherapy, further investigation of this approach should be considered.
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Follicular Center Cell-These describe a subset of lymphocytes which arise within lymphoid follicular germinal centers. Many lymphomas are derived from this subset and are termed follicular center cell lymphomas. These lymphomas may have small or large nuclear size and the nuclei may be cleaved or folded or non-cleaved.
Hodgkin's Cell-General term used to describe variants of Reed-Sternberg cells which lack the diagnostic features of a true Reed-Sternberg cell. These cells often have a single nucleus instead of the binucleated or multinucleated appearance.
Lacunar Cell-A variant of a Reed-Sternberg cell having a clear space around the nucleus. Often present in the Nodular-Sclerosis variant of Hodgkin's lymphoma.
REAL Classification-Revised European-American Lymphoma Classification. This is the latest classification system which incorporates both morphology as well as immunohistochemical and molecular biology data.
Reed-Sternberg Cell (RS Cell)-This is the diagnostic cell of Hodgkin's disease. It is a necessary but not sufficient for the diagnosis. It is a binucleated cell with the two halves having the appearance of a mirror image.
Working Classification-The predecessor classification system to the REAL Classification. It was the first international effort to organize lymphomas into morphologic and prognostic categories. It is still very popular and is often used in coordination with the REAL Classification.
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