Academic literature on the topic 'FISH, LYMPHOMA'

Abstract Cytogenetic analysis is invaluable for the detection of chromosome abnormalities in tumor samples and is the "gold standard" technique (unique in providing a complete overview of the chromosome complement). Cytogenetic studies of lymph node specimens (LN) can be challenging due to progressively smaller biopsies being procured, low viability, and low proliferative rates. Typically, the initial laboratory evaluation of LN includes flow cytometry and/or immunohistochemistry. Due to overlapping immunophenotypic and morphologic features of some lymphomas, these studies can be insufficient to properly classify a lymphoid neoplasm. Interphase FISH is the test most frequently utilized for LN genetic evaluation. Although FISH has higher sensitivity than conventional cytogenetics, there is vast literature on the existence of cytogenetic abnormalities that are not targeted by the FISH probe(s) used in most laboratories. This is predominantly true for CLL/SLL, but it is also seen in other lymphomas, particularly those characterized by variant translocations involving closely related genes, such as mantle cell lymphoma with alternate translocations involving the CCND2 or CCND3 genes, or lymphomas carrying MYC rearrangements where the partner is not IGH (translocation partners other than Ig genes might merit less aggressive therapy). To achieve the same information obtained from an abnormal karyotype, it is usually necessary to perform multiple FISH tests with significantly increased costs. Genomic microarray and sequencing also have limitations. Microarray can only detect DNA unbalances (missing the balanced translocations that characterize most lymphomas). These whole genomic tests cannot detect multiple related clones indicative of clonal evolution, or unrelated clonal populations indicative of distinct lymphoid neoplasms in the same specimen. Successful cytogenetics offers the best visual representation of the whole chromosome complement and often yields information making it unnecessary to perform additional genetic tests. It should be noted that alternative genetic tests are extremely useful for cases with normal chromosome results or those that lack metaphase cells for analysis, as well as those with cytogenetically cryptic rearrangements or mutations. Recent studies indicate that complex karyotypes in lymphomas are, in general, indicative of transformation and/or worse prognosis. In the present study, for example, several follicular lymphoma cases displayed other abnormalities in addition to the typical t(14;18), some of which are known to be associated with transformation, i.e., deletions 1p, 6q, and 10q. We present our experience with 362 LN received over a 15 month period during 2014 and 2015. See Table below. Through correlation with all diagnostic test results from our laboratory, we demonstrate the unique value of cytogenetic evaluation of lymphoid tissues, optimizing diagnostic/prognostic assessment and, thereby, improving patient management/therapeutic decisions, while achieving cost reduction. Table. A) Summary of cases and subdivision based on successful cytogenetics and normal or abnormal flow/morphology versus normal or abnormal cytogenetics; B) Detailed information on the number of the various lymphoid neoplasms included in our study. Total Cases: 362 No metaphases: 67 (19%) With metaphases: 295 (81%) Normal flow/morphology and normal cytogenetics 74 (25%) Abnormal flow and/or morphology 221 (75%) Normal cytogenetics: 59 (27%) Abnormal Cytogenetics: 162 (73%) Table. Final Diagnosis (Abnormal cytogenetic cases) # cases Sex (M/F) # FISH FISH Abnormal/normal FL 56 25/31 43 43/0 SLL/CLL 32 20/12 21 20/1 HGL 14 7/7 0 0 TCL 14 7/7 5 3/2 MZBCL 13 8/5 8 5/3 DH/THL 10 6/4 10 10/0 DLBCL 8 4/4 8 7/1 MCL 7 4/3 5 4/1 CD30+ 2 0/2 2 1/1 NGCL 2 0/2 2 1/1 BL 1 1/0 1 1/0 LPL 1 1/0 0 0 B-ALL/LBL 1 0/1 1 1/0 HL 1 1/0 1 1/0 Totals 162 84/78 107 (66%) 97/10 Abbreviations: FL, follicular lymphoma; SLL/CLL, small lymphocytic lymphoma/chronic lymphocytic leukemia; HGL, high-grade lymphoma; TCL, T-cell lymphoma; MZBCL, marginal zone B-cell lymphoma; DH/THL, double hit/triple-hit lymphoma; DLBCL, diffuse large B-cell lymphoma; MCL, mantle cell lymphoma,; CD30+, CD30-positive large B-cell lymphoma; NGCL, non-germinal center lymphoma; BL, Burkitt lymphoma; LPL, lymphoplasmacytic lymphoma; B-ALL/LBL, B-cell acute lymphoblastic leukemia/lymphoblastic lymphoma; HL, Hodgkin lymphoma. Disclosures No relevant conflicts of interest to declare.

A composite lymphoma is the rare simultaneous occurrence of two or more distinct lymphomas within a single tissue or organ. Herein, we describe a case of a 51-year-old man presenting with a history of lower limb rash, fatigue, and bulky abdominopelvic lymphadenopathy. An excisional left iliac lymph node biopsy was notable for the composite presence of two distinct lymphoid neoplasms, nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL), and follicular lymphoma (FL). Multiplex PCR and FISH analyses failed to demonstrate a t(14;18)(q32;q21) translocation in either composite lymphoma component. A clonal light-chain kappa (V/JC intron-kde) gene rearrangement was detected in the FL component only.

T(11;18)(q21;q21) is the most common structural abnormality in extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) leading to the fusion of the apoptosis inhibitor-2 (API2) gene and the MALT lymphoma-associated translocation (MALT1) gene. In 2 patients with MALT lymphoma of the liver and skin, respectively, t(14;18)(q32;q21) was observed by cytogenetic analysis. Subsequent fluorescence in situ hybridization (FISH) studies disclosed that the immunoglobulin heavy-chain locus (IGH) and the MALT1 gene were rearranged by this translocation. In order to screen a large series of MALT lymphomas for this aberration, a 2-color interphase FISH assay was established. Among a total of 66 cases, t(14;18)(q32;q21) involving IGH and MALT1 was detected in MALT lymphomas of the liver (4 of 4), skin (3 of 11), ocular adnexa (3 of 8), and salivary gland (2 of 11), but did not occur in MALT lymphomas of the stomach (n = 10), intestine (n = 9), lung (n = 7), thyroid (n = 4), or breast (n = 2). In total, 12 of 66 (18%) MALT lymphomas harbored t(14;18)(q32;q21); 7 additional cases of splenic marginal zone lymphoma tested negative. All of the 12 MALT lymphomas featuring the t(14;18)(q32;q21) were negative for t(11;18)(q21;q21) by reverse transcriptase–polymerase chain reaction (RT-PCR). However, trisomy 3 and/or 18 was found in 4 of 12 cases, suggesting that the t(14;18)(q32;q21) does not occur as the sole genetic abnormality. This study identifies IGH as a new translocation partner of MALT1 in MALT lymphomas, which tend to arise frequently at sites other than the gastrointestinal tract and lung. In contrast to t(11;18)(q21;q21)+ MALT lymphomas, those with t(14;18)(q32;q21) may harbor additional genetic abnormalities.

Abstract The differential diagnoses of CD5 positive B-cell lymphoproliferative disorders mainly include chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) and mantle cell lymphoma. Occasionally large cell and marginal zone lymphomas may also be CD5 positive. An accurate diagnosis effects patient management. The classical immunophenotype for chronic lymphocytic leukemia/small lymphocytic lymphoma is CD19/CD5/CD23 positive FMC-7 negative cells with dim CD20 and dim light chain expressions, while mantle cell lymphoma is CD19/CD5/FMC-7 positive with bright CD20 and bright light chain expressions. The diagnosis of mantle cell lymphoma is usually confirmed by either immunostain for cyclin D1 or FISH study for t(11;14). In reality, immunostaining for cyclin D1 can be difficult and may show variable results in different laboratories and FISH study may not be readily available. Generally, when it comes to the diagnosis of lymphoma, immunohistochemical positivity of both CD5 and CD23 is almost pathognomic for chronic lymphocytic leukemia/small lymphocytic lymphoma if no fresh tissue is saved for flow cytometry analysis. Flow cytometry analysis of 44 FISH-confirmed mantle cell lymphomas was reviewed in our lab. Among these, 37 showed the classical immunophenotype of mantle cell lymphoma. However, 7 cases (16%) were positive for both CD5 and CD23. The expression of CD23 varied from dim to bright. When compared to typical CLL, they showed FMC-7 expression and brighter than dim light chain expression. In one case, the light chain expression was dim. In conclusion, CD23 expression which was thought to be a specific marker for CLL/SLL may also be seen with mantle cell lymphoma. Although FMC-7 expression is seen in all CD23 positive mantle cell lymphomas, bright light chain expression is not universal. We recommend that FISH or immunohistochemical studies for cyclin D1 be performed on CD5/CD19 clonal B cell proliferations with CD23 expression if morphology or immunophenotype is atypical for CLL/SLL.

Abstract Introduction: Composite lymphomas involving Hodgkin lymphoma (HL) and non-Hodgkin lymphomas (CLL, DLBCL, FL, MZL, MCL, T-NHL) are relatively rare but are increasingly frequently diagnosed. This may be a function of the change in diagnostic practice, the more varied and increased treatment of the presenting disease or simply reflect better monitoring post treatment with re-biopsy of lymph nodes. Composite lymphoma is defined as the synchronous and metachronous development of two or more lymphomas in the same patient. The mechanism of pathogenesis underlying its occurrence is not clearly established and in particular the relationship between entities when HL is diagnosed a number of years following successful treatment for follicular lymphoma (FL). Methods: We identified 48 patients with a diagnosis of Hodgkin lymphoma at our centre between January 2005 and June 2014 who had a previous or concurrent diagnosis of another lymphoproliferative disorder. Diagnoses included CLL = 14; follicular lymphoma = 13; DLBCL = 8; mantle cell lymphoma = 1; marginal zone lymphoma = 5; T-cell lymphoma = 3; not specified = 3 and a single patient had CLL, MZL and most recently MCL. The diagnosis of Hodgkin lymphoma was confirmed on the tissue biopsy by using a standard panel of immunohistochemistry markers (CD3, CD20, CD30, IRF4, TARC, CD79, LMP1, BCL6, BOB1, OCT2). This study is focussed on the 13 cases with follicular lymphoma. The aim of the study was to identify a relationship between the Hodgkin lymphoma and the follicular lymphoma using interphase FISH studies to look for identical chromosomal translocations. Results: 8/13 patients had follicular lymphoma diagnosed on an earlier tissue biopsy, range 1-9 years prior to the diagnosis of Hodgkin lymphoma; 7/8 of these FL cases had bone marrow staging carried out at presentation and 5/7 had involvement by FL. 5/13 were newly presenting patients with composite lymphoma; 3/5 had a staging marrow which in each case showed evidence of follicular lymphoma alone with no evidence of Hodgkin lymphoma. In total, 7/13 patients fit the criteria for composite lymphoma with Hodgkin lymphoma and follicular lymphoma occupying distinct zones within the same tissue biopsy. The remaining 6/13 patients show no evidence of follicular lymphoma in the current biopsy and are indistinguishable from de novo presentation of Hodgkin lymphoma. Interphase FISH was used to assess for a genetic relationship between the disease entities. 3µ sections were cut from formalin fixed paraffin processed tissue, using the same block where possible as the H&E and immunohistochemistry. All cases were independently assessed by two experienced scientists with knowledge of histology and FISH reporting on thin sections. 8/13 cases had suitable samples for FISH, the remaining 5 cases had insufficient material remaining in the block. Commercial probes for BCL2 ‘Split-Signal’ (Abbott/Vysis or Dako) were used and FISH results were examined using a Zeiss microscope with MetaSystem image capture. All 8 cases showed BCL2 gene rearrangement in the Reed-Sternberg (RS) cells (5/8 cases were de novo group of Hodgkin lymphoma with no morphological or phenotypic evidence of FL in the tissue and 3/8 were composite lymphoma with distinct zoning). One rare case contained RS cells in a dab/imprint preparation made from the fresh tissue, FICTION technique was carried out on this case combining CD30 immunofluorescent staining with FISH for BCL2 gene rearrangement confirming RS cell type with BCL2 gene rearrangement. CONCLUSIONS The identification of BCL2 gene rearrangement in RS cells, the hallmark cell of Hodgkin lymphoma, in this series of composite lymphomas suggests a relationship between the B cells of follicular lymphoma and the RS cells of HL. The presence of the same chromosomal abnormalities identified in more than one lymphoma cell type indicates the same clonal cell of origin. Disclosures Jack: Roche: Research Funding; Genentech: Collaboration, Collaboration Other.

Abstract Context.—Molecular genetic analyses have been predicted to improve the diagnostic accuracy of fine-needle aspiration of B-cell non-Hodgkin lymphoma. Objective.—To determine the value of routine molecular genetic assays, polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH), in the diagnosis of B-cell non-Hodgkin lymphoma by fine-needle aspiration (FNA). Design.—A multiparametric method, including cytology, flow cytometry, PCR, and FISH, was prospectively evaluated in the diagnosis of B-cell non-Hodgkin lymphoma by FNA. Aspirates from 30 consecutive patients with suspected hematolymphoid malignancies were collected. All aspirates were triaged through a uniform program including cell-size analysis, B- and T-cell clonality studies, flow cytometric immunophenotyping, and bcl-1 and bcl-2 gene rearrangements by PCR and FISH. After completion of FNA evaluations, FNA results were compared with diagnoses from prior or subsequent surgical biopsies. Results.—Monoclonal B-cell populations were detected in 18 of 20 B-cell non-Hodgkin lymphomas by flow cytometry and PCR. bcl-1 gene rearrangement was detected in 2 of 2 cases of mantle cell lymphoma. bcl-2 rearrangement was detected in 5 cases including 4 of 4 low-grade follicular lymphomas and 1 transformed follicular lymphoma. By incorporating the results of molecular genetic and ancillary diagnostics, a definitive classification was reached in 12 cases of B-cell non-Hodgkin lymphoma by FNA, including all cases of low-grade follicular lymphoma (4/4) and mantle cell lymphoma (2/2) and approximately 50% of small lymphocytic lymphoma (2/4) and large B-cell lymphoma (4/8). Ten of the 12 cases with a final classification reached by FNA had either prior or follow-up surgical biopsies, and all 10 cases showed agreement between the diagnoses rendered on FNA and surgical biopsies. Conclusions.—With proper handling and management of specimens, FNA can routinely provide samples adequate for molecular genetic studies, in addition to cytomorphology and flow cytometry, making it possible to consistently render accurate and definitive diagnoses in a subset of B-cell non-Hodgkin lymphomas. By incorporating FISH and PCR methods, FNA may assume an expanded role for the primary diagnosis of B-cell non-Hodgkin lymphoma.

Abstract Abstract 4722 Fluorescence in situ hybridization (FISH) panel for detecting lymphoid neoplasms in interphase nuclei currently used at most of the laboratories initially utilizes the IGH (14q32.3) locus specific probe and based on negative or positive results obtained, additional testing is done with either the BCL6 (3q27), MYC (8q24) and MALT1 (18q21) locus specific probes, or the MYC/IGH (8q24/14q32.3), CCND1/IGH (11q13/14q32.3) and IGH/BCL2 (14q32.3/18q22) locus specific probes, respectively. Although these probes detect a large number of lymphoma-related abnormalities, approximately 10% of cases are still resulted negative by this FISH panel, as well as cytogenetics analysis on lymphoid mitogens stimulated bone marrow/peripheral blood cultures, but positive by hematopathology. Apparently, a need exists for either an expansion of this FISH panel or development of a reflex FISH panel to be used only when the current FISH panel fails to detect any abnormality in such cases. We selected three additional probes, TCR a/d, MYB and ALK to test further in a pilot study including twenty-five such patients, based on a high number of studies describing involvement of specific loci in gene rearrangements in lymphoid neoplasms. TCR a/d (14q11) locus is frequently involved in gene rearrangements in T-cell lymphomas and leukemias. These rearrangements include t(1;14)(q32;q11), t(1;14)(q34;q11), t(7;14)(p15;q11), t(7;14)(q34-35;q11), t(8;14)(q24;q11), t(10;14)(q24;q11), t(11;14)(p13;q11), t(11;14) (p15;q11) and inv(14)(q11;q32)/t(14;14)(q11;q32), among others. MYB (6q23) locus shows loss of heterozygosity in a high proportion of patients with peripheral T-cell and NK/T-cell lymphomas. ALK (2p23) locus has also been well documented as involved in gene fusion with multiple partner loci, including 1q21, 2q11-13, 2q35, 3q21, 5q35, 17q23, 19p13.1 and Xq11-12 in Ki-1-positive anaplastic large cell lymphoma, a subtype of non-Hodgkin lymphoma involving Ki-1 antigen. A known negative control for each probe and a known MYB-positive control were used in a blind-coded set-up with the criteria of a positive result with abnormal probe signal pattern in at least 5% (10/200) of cells examined with the validated standard FISH protocols. This experimental reflex panel succeeded in detecting abnormalities in three of the twenty-five (12%) patients included in this pilot study. Two cases were positive for the loss of heterozygosity for the MYB locus, while one case was positive for involvement of TCR a/d locus in a translocation. The ALK probe did not detect any additional abnormality, but none of the patients had a hematopathology diagnosis of Ki-1+ anaplastic large cell lymphoma. Only four of the twenty-five patients studied had a hematopathology diagnosis of a T-cell disorder. Further, some of the patients included in this study had a low percentage of cells found abnormal by hematopathology. These factors could have affected the observed rate of abnormalities by this reflex FISH panel. Still, with 12% of patients showing abnormalities with this reflex FISH panel, this pilot study clearly demonstrates the usefulness of incorporation of this reflex FISH panel in the standard protocols for FISH studies in lymphoid neoplasms. We plan to continue investigation on additional patients meeting the criteria for inclusion in this study. Disclosures: No relevant conflicts of interest to declare.

Splenic lymphoma may be primary or secondary. Primary splenic lymphoma’s are rare and usually of follicular cell origin representing <1% of Non-Hodgkin’s Lymphoma’s. Most are secondary with 35% representing Marginal Cell sub-type with the rest being Diffuse Large B-Cell Lymphoma’s. Unlike the uniformly aggressive clinical course of Diffuse Large B-Cell Lymphoma’s, biological behavior of Primary Splenic CD10-Positive Small B-Cell Lymphoma/Follicular Lymphoma remains less well defined. We present here a solitary splenic mass confirmed as Primary Splenic CD10-Positive Small B-Cell Lymphoma/Follicular Lymphoma after a diagnostic splenectomy. Biopsy revealed monomorphic small lymphoid cells with low grade mitotic activity. Flow cytometry showed a lambda restricted population of B-Cells displaying dim CD19 and CD10. The cells were negative for CD5, CD11c, and CD103. FISH was negative for IGH/BCL2 fusion unlike nodal Follicular Lymphoma’s which are usually positive for this translocation. Evidence from this case and a review of literature support the finding that Primary Splenic CD10-Positive Small B-Cell Lymphoma/Follicular Lymphoma is less likely to have the classic IGH-BCL2 fusion and the associated chromosomal 14;18 translocation. This profile is associated with less aggressive clinical behavior even when histopathology represents a high-grade pattern. In such cases splenectomy alone is adequate for localized disease when negative for IGH/BCL2 fusion regardless of histological grade.

Abstract The translocation of chromosome 11, long arm, region 2, band 1, to chromosome 18, long arm, region 2, band 1 (t(11;18)(q21;q21)) represents a recurrent chromosomal abnormality in extranodal marginal zone B-cell lymphoma (MZBCL) of mucosa-associated lymphoid tissue (MALT) type and leads to a fusion of the apoptosis inhibitor-2 (API2) gene on chromosome 11 and the MALT lymphoma-associated translocation (MLT) gene on chromosome 18. A 2-color fluorescence in situ hybridization (FISH) assay, which can be used for the detection of t(11;18) in interphase nuclei and metaphase chromosomes on fresh and archival tumor tissue, was developed. The P1 artificial chromosome (PAC) clone located immediately telomeric to the MLT gene and the PAC clone spanning the API2 gene were differentially labeled and used to visualize the derivative chromosome 11 resulting from t(11;18), as evident by the overlapping or juxtaposed red and green fluorescent signals. The assay was applied to interphase nuclei of 20 cases with nonmalignant conditions and 122 B-cell non-Hodgkin's lymphomas (NHLs). The latter group comprised 20 cases of nodal follicle center cell lymphoma and diffuse large B-cell NHL, 10 cases of gastric diffuse large B-cell lymphoma, 10 cases of hairy cell leukemia, and 82 cases of MZBCL (41 extranodal from various locations, 19 nodal, and 22 splenic MZBCL) including 35 cases with an abnormal karyotype, 2 of which revealed t(11;18). By interphase FISH, t(11;18) was detected in 8 gastrointestinal low-grade MALT-type lymphomas including the 2 cytogenetically t(11;18)+ cases. In the 8 t(11;18)+ cases, the FISH results were confirmed by reverse transcriptase–polymerase chain reaction (RT-PCR) usingAPI2 and MLT specific primers. Our results indicate that t(11;18)(q21;q21) specifically characterizes a subgroup of low-grade MZBCL of the MALT-type and that the FISH assay described here is a highly specific and rapid test for the detection of this translocation.

The translocation of chromosome 11, long arm, region 2, band 1, to chromosome 18, long arm, region 2, band 1 (t(11;18)(q21;q21)) represents a recurrent chromosomal abnormality in extranodal marginal zone B-cell lymphoma (MZBCL) of mucosa-associated lymphoid tissue (MALT) type and leads to a fusion of the apoptosis inhibitor-2 (API2) gene on chromosome 11 and the MALT lymphoma-associated translocation (MLT) gene on chromosome 18. A 2-color fluorescence in situ hybridization (FISH) assay, which can be used for the detection of t(11;18) in interphase nuclei and metaphase chromosomes on fresh and archival tumor tissue, was developed. The P1 artificial chromosome (PAC) clone located immediately telomeric to the MLT gene and the PAC clone spanning the API2 gene were differentially labeled and used to visualize the derivative chromosome 11 resulting from t(11;18), as evident by the overlapping or juxtaposed red and green fluorescent signals. The assay was applied to interphase nuclei of 20 cases with nonmalignant conditions and 122 B-cell non-Hodgkin's lymphomas (NHLs). The latter group comprised 20 cases of nodal follicle center cell lymphoma and diffuse large B-cell NHL, 10 cases of gastric diffuse large B-cell lymphoma, 10 cases of hairy cell leukemia, and 82 cases of MZBCL (41 extranodal from various locations, 19 nodal, and 22 splenic MZBCL) including 35 cases with an abnormal karyotype, 2 of which revealed t(11;18). By interphase FISH, t(11;18) was detected in 8 gastrointestinal low-grade MALT-type lymphomas including the 2 cytogenetically t(11;18)+ cases. In the 8 t(11;18)+ cases, the FISH results were confirmed by reverse transcriptase–polymerase chain reaction (RT-PCR) usingAPI2 and MLT specific primers. Our results indicate that t(11;18)(q21;q21) specifically characterizes a subgroup of low-grade MZBCL of the MALT-type and that the FISH assay described here is a highly specific and rapid test for the detection of this translocation.