Flow cytometry immunophenotyping of fine-needle aspiration specimens: utility in the diagnosis and classification of non-Hodgkin lymphomas

Barrena S, Almeida J, García‐Macias M D C, López A, Rasillo A, Sayagués J M, Rivas R A, Gutiérrez M L, Ciudad J, Flores T, Balanzategui A, Caballero M D & Orfao A
(2011) Histopathology  58 , 906–918
Flow cytometry immunophenotyping of fine‐needle aspiration specimens: utility in the diagnosis and classification of non‐Hodgkin lymphomas Aims: To establish the utility of flow cytometry (FCM) for screening and diagnosis of B cell non‐Hodgkin lymphoma (B‐NHL) from lymphoid tissue samples obtained by fine‐needle aspiration (FNA). Methods and results: We compared prospectively FCM versus cytology/histology analysis of FNA samples for the diagnostic screening and further World Health Organization (WHO) subclassification of B‐NHL. FCM and cytology showed a high degree of agreement (93%); however, diagnosis of reactive processes (RP), B‐NHL and T‐NHL by FCM showed higher sensitivity than cytology (92–100% versus 64–94%, respectively), without false positive NHL cases. The antibody combination used did not allow a positive diagnosis of Hodgkin lymphoma as distinct from a RP. A high concordance rate was found between FCM and histopathology (74%) in subtyping B‐NHL. In this regard, mantle‐cell lymphoma and chronic lymphocytic leukaemia/small lymphocytic lymphoma showed the highest degree of agreement (100% concordant rates). In turn, FCM showed higher sensitivity/specificity in classifying follicular lymphoma (FL) and large B cell lymphomas, while the opposite occurred for marginal‐zone and lymphoplasmacytic lymphomas. Conclusions: FCM enhances the diagnostic ability of FNA cytology, playing a crucial role in a rapid and accurate differential diagnosis between RP, B‐NHL and T‐NHL. In addition, immunophenotyping of FNA samples contributes to a more precise subclassification of B‐NHL when combined with histopathology and genetic/molecular data.     

The diagnostic approach to fine‐needle aspiration of malignant lymphoma: Using cytomorphology and immunocytochemistry with cell transfer method

Fine‐needle aspiration (FNA) cytology is generally considered to be the screening tool for lymphoproliferative lesions. The differential and decisive diagnosis, however, of malignant lymphoma from benign reactive hyperplasia by FNA cytology is sometimes challenging. The diagnostic features compatible with lymphoma as opposed to reactive hyperplasia in FNA cytology were investigated with 31 cases of lymphoma and 31 cases of reactive hyperplasia, and immunocytochemistry with cell transfer method was additionally applied to FNA cytology. The predominance of large lymphocytes, the clustering of large lymphocytes, the presence of markedly large and/or highly pleomorphic cells, the presence of apoptotic and/or necrotic cell debris were considered characteristics of lymphomas, whereas the predominance of small lymphocytes and the presence of histiocytes were considered characteristics of reactive hyperplasia. Using these cytomorphologic characteristics, the diagnostic accuracy for malignant lymphoma in FNA cytology had a sensitivity of 80.6% and a specificity of 100%. By cell transfer method, one of Papanicolaou‐stained slides could be used in immunocytochemistry for several markers. Using such methods, sensitivity of FNA cytology for lymphoma was upgraded to 100%, and decisive diagnoses of diffuse large B‐cell lymphoma, Burkitt lymphoma, low grade B‐cell lymphoma, T‐ or NK‐cell non‐Hodgkin lymphoma (NHL), or Hodgkin lymphoma was possible. Differential diagnosis of malignant lymphoma from reactive hyperplasia, and decisive diagnoses of high, and low grade B‐cell NHL, T‐ or NK‐cell NHL, and HL could be possible by FNA cytology with cytomorphology in conjunction with immunocytochemistry using cell transfer method. Diagn. Cytopathol. 2014;42:671–679. © 2014 Wiley Periodicals, Inc.     

Two cases of primary non‐Hodgkin's lymphoma of female breast: Role of fine‐needle aspiration cytology and cell‐block immunohistochemistry

Primary breast lymphoma (PBL) is an uncommon neoplastic condition. Though HIV‐infection is a known risk factor for the development of extranodal lymphomas, mammary involvement is still a rarity. Radiologically, PBL appears as well circumscribed, heteroechoic, noncalcifying mass. Fine‐needle aspiration cytology (FNAC) is commonly used to diagnose this neoplasm; however, subcategorization requires immunophenotypic characterization of the neoplastic cells. Herein, we report two cases of PBL, including a HIV‐infected lady; in both the cases FNAC expressed features of non‐Hodgkin's lymphoma. Finally, immunohistochemistry on cell‐block with CD20 diagnosed both the cases as diffuse large B‐cell lymphoma (DLBCL). Diagn. Cytopathol. 2016;44:235–240. © 2015 Wiley Periodicals, Inc.     

Hodgkin's lymphoma: Diagnostic difficulties in fine‐needle aspiration cytology

It is commonly believed that cytodiagnosis of Hodgkin's lymphoma (HL) is much easier than that of non‐Hodgkin lymphoma (NHL). However, recognition of certain NHL subtypes with Reed‐Sternberg (R‐S)‐like cells and results of immunohistochemical studies point to the contrary. To study the limitations of cytology in diagnosis of HL, fine‐needle aspiration (FNA) smears of 130 lymphoma or suspected lymphoma cases were reviewed. Initial and reviewed cytodiagnoses were compared with histopathology in 89 cases. Immunocytochemical and immunohistochemical studies were performed in 56 and 59 cases, respectively. Among histologically diagnosed HL cases, definitive cytodiagnosis of HL (initial as well as reviewed) was significantly less frequent than cytodiagnosis of NHL among histologically diagnosed NHL cases (P = 0.0328 and = 0.0001, respectively). On the other hand, cytologically diagnosed HL/NHL cases were significantly more frequent in the former group (P = 0.0001 and = 0.0018, respectively). ALCL and TCRBCL were the two NHL subtypes which created confusion with HL in FNA smears. Twenty‐one cytohistological concordant HL cases and equal number of discordant cases were compared. When compared with discordant group, the patients in concordant group were significantly younger (P = 0.045). Hodgkin/Hodgkin‐like cells and typical R‐S cells were significantly more frequent in FNA smears of the concordant group (P = 0.0478 and = 0.0431, respectively). Immunocytochemical and immunohistochemical studies showed good correlation with histological diagnosis of HL. It is suggested that proper interpretation of cytologic features, together with use of immunocytochemical parameters can help in reducing the margin of error in cytodiagnois of HL. Diagn. Cytopathol. 2009. © 2009 Wiley‐Liss, Inc.     

Non‐Hodgkin's lymphoma presenting as breast masses: A series of 10 cases diagnosed on FNAC

Primary breast lymphoma (PBL) is a rare disease, which comprises 0.04–0.53% of all primary malignant tumors of the breast. The most frequent histological subtype is diffuse large B‐cell type (DLBCL) (40–70%). Differentiation of PBLs from other breast tumors such as poorly differentiated carcinomas and lobular carcinoma may at times be difficult on cytomorphology alone. An audit of breast lymphomas diagnosed on fine needle aspiration cytology (FNAC) over a period of 9 years (2001–2009) was performed. Ten cases were retrieved and the cytomorphology was reviewed along with immunochemistry (IHC), flow cytometry as well as histopathology, wherever available. The age of patients ranged from 17 to 83 years. Eight cases were diagnosed as non‐Hodgkin's lymphoma, high‐grade on FNAC. Histopathology was available in four of these cases and cell block was available in one case. Lymphoid cells were positive for leukocyte‐common antigen (LCA) and CD20 and negative for CD3 in these cases. The same was confirmed by flow cytometry on aspirated material in one case. A diagnosis of DLBCL was offered in these five cases. One case was a low‐grade NHL and another case was a young male, a known case of acute leukemia and had leukemic infiltration in the breast lump. We wish to emphasize the potential importance of FNAC in breast lymphoma and the same can be helpful to avoid unnecessary surgery in these cases. The differential diagnostic entities have been discussed. IHC and flow cytometry can be performed on the aspirated material and provide valuable information. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

Peripheral T‐cell and NK‐cell lymphomas and their mimics; taking a step forward – report on the lymphoma workshop of the XVIth meeting of the European Association for Haematopathology and the Society for Hematopathology

Mature T‐cell and T/NK‐cell neoplasms are both uncommon and heterogeneous, among the broad category of non‐Hodgkin lymphomas. Owing to the lack of specific genetic alterations in the vast majority, most currently defined entities show overlapping morphological and immunophenotypic features, and therefore pose a challenge to the diagnostic pathologist. In the light of recent immunophenotypic, cytogenetic and molecular genetics advances in the field of T‐cell and T/NK‐cell lymphomas, the focus of the lymphoma workshop of the European Association for Haematopathology/Society for Hematopathology meeting in Lisbon, Portugal, in October 2012 was to refine existing diagnostic criteria and clarify the borders between overlapping entities. The panel reviewed over 200 submitted cases, which were grouped into five categories: (i) angioimmunoblastic T‐cell lymphoma and T‐follicular‐helper‐cell‐associated lymphomas; (ii) CD30‐positive T‐cell lymphomas/lymphoproliferative diseases; (iii) extranodal T‐cell and NK‐cell neoplasms; (iv) EBV‐associated T‐cell/NK‐cell lymphomas/lymphoproliferative diseases; and (v) peripheral T‐cell lymphoma, not otherwise specified, post‐transplant lymphoproliferative disorders, and mimics. This report summarizes the discussions and conclusions of the workshop, which question current diagnostic criteria and provide recommendations for refining existing classifications.     

Reed‐sternberg cells in breast FNA of a patient with left breast mass

Hodgkin's lymphoma is a potentially curable malignancy of the lymphatic system characterized by a variable number of scattered and large mononucleated and multinucleated tumor cells, the Hodgkin and Reed‐Sternberg cells residing in an abundant heterogeneous admixture of non‐neoplastic inflammatory cells. It represents approximately 30% of all lymphomas according to the World Health Organization (WHO). Patients with Hodgkin's lymphoma typically present with painless peripheral adenopathy, fever, night sweats, and weight loss. We report a rare case of Hodgkin's lymphoma presented as a breast mass in a 23‐year‐old woman diagnosed on fine needle aspiration (FNA). At presentation, she had no B symptoms, or palpable lymphadenopathy. Diagn. Cytopathol. 2010;38:663–668. © 2010 Wiley‐Liss, Inc.     

Application of the BIOMED‐2 standardized primer system for the diagnosis of primary gastric B‐cell non‐Hodgkin's lymphoma

Primary gastric B‐cell non‐Hodgkin's lymphoma (B‐NHL) has a similar morphocytological presentation to severe chronic gastritis, which complicates the pathological diagnosis of this disease. To investigate the practicality and utility of the BIOMED‐2 standardized primer system for the diagnosis of primary gastric B‐NHL from endoscopic biopsy specimens, we selected 65 cases of archived paraffin‐embedded primary gastric B‐NHL specimens as well as 27 cases of severe chronic gastritis samples to serve as a negative control group. The positivity rates of immunoglobulin heavy chain (IgH) gene rearrangements detected by the BIOMED‐2 standardized primer system for the two groups were 86.4% and 12.0%, respectively, which are significantly different (p < 0.05). Importantly, the combined detection of the five groups of the IgH primer system increased the detection rate of B‐NHL. These findings indicate that the BIOMED‐2 standardized primer system is suitable for formalin‐fixed and paraffin‐embedded (FFPE) specimens and is valuable as a secondary diagnostic tool for primary gastric B‐NHL as well as the differential diagnosis of severe chronic gastritis.     

Primary diagnosis and REAL/WHO classification of non-Hodgkin's lymphoma by fine-needle aspiration: cytomorphologic and immunophenotypic approach

The Revised European American lymphoma (REAL) and World Health Organization (WHO) classification of non-Hodgkin's lymphoma (NHL) relies on the constellation of cytologic, phenotypic, genotypic, and clinical characteristics of NHL. For the most part, the classification does not rely on architectural pattern for classification of neoplasms. This classification makes it possible to diagnose and classify lymphomas by fine-needle aspiration (FNA). In this study, we attempted to evaluate the accuracy of FNA in diagnosing and classifying NHL within the context of the REAL/WHO classifications. Cases included only those in which FNA was the primary diagnosis, followed by a surgical biopsy for confirmation. Flow cytometry (FCM) for phenotyping was carried out whenever material was available. Two groups of pathologists were identified. Group A consisted of pathologists with background training in cytopathology and/or hematopathology (three pathologists). Group B consisted of experienced surgical pathologists with no training in cytopathology and/or hematopathology (four pathologists). Seventy-four cases were included in the study. FCM phenotyping was performed in 53 cases (71%). Large cell lymphoma constituted 63% of the cases. The remaining lymphomas included Burkitt's, small lymphocytic, lymphoblastic, follicle center cell, Ki-1, mantle cell, marginal zone, and natural killer cell lymphoma. The diagnosis of lymphoma was rendered for all cases. The correct classification was seen in 63% of the cases. Classification was more accurate in immunophenotyped than in nonimmunophenotyped cases (84% vs 33%; P = 0.00004). Group A pathologists showed higher incidence of proper classification than group B (80% vs 56%; P = 0.046). The diagnosis and classification of NHL can be achieved in a large number of cases on FNA material. This accuracy can be increased if cytomorphologic criteria are established for different entities of NHL aided by FCM for phenotyping.     

Pulmonary non‐Hodgkin's lymphoma (NHL) of diffuse large B‐cell type with simultaneous humeral involvement in a young lady: An uncommon presentation with cytologic implications

A bronchogenic carcinoma, almost invariably, presents as a lung mass. Primary pulmonary lymphomas are rare. We report an unusual case of a pulmonary non‐Hodgkin's lymphoma (NHL) with simultaneous involvement of the right humerus in a 37 year old lady. Bronchial lavage smears showed atypical cells with irregular nuclear membranes raising a suspicion of a hematolymphoid tumor, over a small cell carcinoma that was the closest differential diagnosis. Biopsy from the lung mass and from the lesion in the humerus showed an identical malignant round cell tumor with prominent apoptosis. On immunohistochemistry (IHC), tumor cells were diffusely positive for leukocyte common antigen (LCA), CD20 and MIB1 (70%), while negative for cytokeratin (CK), epithelial membrane antigen (EMA) synaptophysin, chromogranin, neuron specific enolase (NSE), CD3, and CD10. Diagnosis of a pulmonary NHL of diffuse large B‐cell type with involvement of the humerus was formed. The case is presented to create an index of suspicion for the possibility of a NHL on respiratory samples, while dealing with small round cells with irregular nuclear membranes. IHC is necessary to confirm he diagnosis. A simultaneous association in the humerus in our case makes it unusual. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.     

AA‐type amyloidosis in association with non‐Hodgkin's lymphoma following CMV viremia: Autopsy case

Amyloidosis in non‐Hodgkin's lymphoma (NHL) is known to be of the AL type, and AA‐type amyloidosis in NHL is extremely rare. Herein is reported an autopsy case of follicular lymphoma that transformed to diffuse large B‐cell lymphoma (DLBCL) in a relapse associated with systemic AA amyloidosis. CMV infection in an immunocompromised state with chemotherapy against DLBCL may have been involved in amyloid accumulation. The serum amyloid A (SAA)1 gene polymorphism, SAA1.2/1.3, might have also been another factor in this case, considering the risk of AA amyloidosis in Japanese patients with rheumatoid arthritis.     

Primary pancreatic diffuse large B‐cell lymphoma diagnosed by endoscopic ultrasound guided FNAC: A rare entity

Primary pancreatic lymphoma (PPL) is an uncommon neoplasm which can clinico‐radiologically mimic carcinoma. But the management of these patients differs from that of a carcinoma. Endoscopic ultrasound (EUS) guided fine‐needle aspiration (FNA) serves as a potential tool to identify pancreatic lymphomas and thus prevent an invasive diagnostic test. This case report describes the presentation and diagnosis of primary pancreatic lymphoma. A 37‐year‐old female presented with nausea, vomiting with signs of icterus and elevated liver function test and Bilirubin. Abdominal computed tomography (CT) revealed a hypodense lesion in the head of the pancreas. EUS guided FNA was performed and cytological material was collected. The lesion was diagnosed as Non‐Hodgkin Lymphoma (NHL) and subtyped as diffuse large B‐cell lymphoma‐germinal centre (DLBCL‐GCB) base on immunohistochemistry on cell block. The patient was started on rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone (RCHOP) regimen. EUS guided FNA along with ROSE, cell bock, and immunocytochemistry helps in the diagnosis of primary pancreatic lymphoma.     

The B‐cell receptor in control of tumor B‐cell fitness: Biology and clinical relevance

Surface expression of a functional B cell antigen receptor (BCR) is essential for the survival and proliferation of mature B cells. Most types of B‐cell lymphoproliferative disorders retain surface BCR expression, including B‐cell non‐Hodgkin lymphomas (B‐NHL) and chronic lymphocytic leukemia (CLL). Targeting BCR effectors in B‐NHL cell lines in vitro has indicated that this signaling axis is crucial for malignant B cell growth. This has led to the development of inhibitors of BCR signaling, which are currently used for the treatment of CLL and several B‐NHL subtypes. Recent studies based on conditional BCR inactivation in a MYC‐driven mouse B‐cell lymphoma model have revisited the role of the BCR in MYC‐expressing tumor B cells. Indeed, lymphoma cells losing BCR expression continue to grow unless subjected to competition with their BCR‐expressing counterparts, which causes their elimination. Here, we discuss the molecular nature of the fitness signal delivered by the BCR to MYC‐expressing malignant B cells, ensuring their preferential persistence within a rapidly expanding tumor population. We also review growing evidence of Ig‐negative cases belonging to several B‐NHL subtypes and CLL, and discuss the clinical implications of these findings in relation to an emerging picture of clinical resistances to anti‐BCR therapies.     

Value and limitations of fine-needle aspiration cytology in diagnosis and classification of lymphomas: A review.

The fine needle aspiration (FNA) cytologic diagnosis of non-Hodgkin's lymphoma (NHL) depends upon finding a relatively monotonous population of lymphoid cells in smears. Lymphomas have successfully been classified by FNA cytology following the prevalent histologic classifications. The success rate of FNA cytology ranges from 80%-90% in diagnosis of NHL and from 67.5%-86% in its subtyping. The cytodiagnosis of Hodgkin's disease (HD) depends upon demonstration of Reed-Sternberg cells or Hodgkin's cells amongst appropriate reactive cell components. The diagnostic accuracy of FNA cytology for HD has also been invariably high (>85%). Yet, the role of cytology in primary diagnosis, subclassification and management of patients with lymphoma remains controversial. The differential diagnostic problems for NHL include a group of small round cell tumors, nonlymphoid acute leukemias and HD. Reservations have been expressed regarding the efficacy of cytology in separating florid reactive hyperplasia from low-grade malignant lymphoma. The reported cytodiagnostic accuracy for follicular lymphomas and nodular sclerosis type of HD is less compared to other subtypes of NHL and HD respectively since nodular pattern and sclerosis are strict histologic criteria which can not be appreciated in cytologic preparations. Entities like atypical lymphoproliferative disorders, peripheral T-cell lymphomas and Ki-1 positive anaplastic large cell lymphomas pose diagnostic challenges to cytologists. Despite these limitations, FNA cytology remains the first line of investigations (screening test) used in cases of lymphadenopathy. Besides initial diagnosis of lymphoma, it helps in detection of residual disease, recurrences and progression of low-grade to high-grade lymphoma, and helps in staging the disease. Availability of prior FNA cytology report facilitates the histologic diagnosis and classification of NHL. Various special ancillary techniques are now being performed on lymph node aspirates to diagnose lymphoma versus other malignancies, and to decide the functional character of lymphomas and their clonal nature. Diagn. Cytopathol. 1999;21:240-249.     

Anaplastic lymphoma kinase positive large B‐cell lymphoma: Literature review and report of an endoscopic fine needle aspiration case with tigroid backgrounds mimicking seminoma

Anaplastic lymphoma kinase‐positive large B‐cell lymphoma (ALK+ LBCL) is a rare distinct type of non‐Hodgkin's lymphoma that arises in association with alterations of the ALK gene. This distinct disease entity is typically associated with an aggressive clinical course and appears in light microscopic preparations as a monomorphic population of large, immunoblast‐like cells. In this report, we describe a case of ALK+ LBCL diagnosed by transgastric endoscopic ultrasound‐guided fine needle aspiration (EUS FNA) of splenic hilar lymph nodes. Modified Giemsa stained direct smears from the FNA sample demonstrated large lesional cells with foamy cytoplasm and macronucleoli admixed with small lymphocytes in tigroid backgrounds, mimicking the cytologic appearance of seminoma. Ancillary immunohistochemical studies subsequently confirmed the diagnosis of ALK+ LBCL with the lesional cells being immunoreactive for CD138, VS38c, MUM1, ALK1, and lambda light chain. The cohesiveness of the cells, the cellular morphology, and the tigroid backgrounds were all pitfalls for accurate diagnosis of this rare specific type of lymphoid malignancy by cytology. To our knowledge this is the first case report detailing the diagnosis of ALK+ LBCL by EUS FNA and the first report describing a glycogen‐rich tigroid background in direct FNA smears. Establishing a refined diagnosis in cases of this rare form of LBCL is necessary, as therapies targeting ALK may be of value in clinical management. Diagn. Cytopathol. 2017;45:148–155. © 2016 Wiley Periodicals, Inc.     

A primary nipple lymphoma diagnosed by a modified fine‐needle aspiration method

Primary breast lymphomas are uncommon. All reported primary breast lymphomas were in the breast parenchyma. Here we reported the first case of primary nipple lymphoma in a 76‐year‐old woman initially diagnosed using a modified fine‐needle aspiration method. The aspirated material by this method had yielded adequate material for both cytomorphologic and flow cytometric analysis, as well as for molecular analysis of light chain rearrangement. In smears the atypical lymphocytes were predominantly middle‐sized with irregular nuclei. Scattered large centroblast or immunoblast‐like cells, a few reactive lymphocytes, histocytes, and few plasma cells were also observed. These findings suggested a low‐grade lymphoma that was further confirmed by flow cytometry (CD19+/CD3?, positive for cytoplasmic kappa light chain but negative for lambda light chain) and molecular analysis (monoclonal rearrangement of immunoglobulin kappa chain). Immunohistochemical stains performed on the excised specimen showed that the tumor cells were positive for CD20 and CD79a but negative for cytokeratin, CD3, CD5, CD10, CD23, CD43, CD45RO, bcl‐6, and cyclin‐D1. The Ki‐67 proliferation index was less than 20%. Taking these together, the case was diagnosed as a primary MALT lymphoma of the nipple. FNA usually provides a better cell morphology than tissue sections, but pathologists have to face the sampling error and lack of immunophenotype information when subtyping lymphoma issues using FNA. With the help of flow cytometry and molecular analysis, more and more trials haveproved the accuracy of FNA in diagnosis of lymphomas. Therefore, FNA could play an informative and diagnostic role indiagnosis of lymphoma. Diagn. Cytopathol. 2012. © 2011 Wiley Periodicals, Inc.     

Diagnostic impact of core-needle biopsy on fine-needle aspiration of non-Hodgkin lymphoma

We retrospectively reviewed 74 fine-needle aspiration (FNA) cases of presumptive non-Hodgkin lymphoma (NHL). All the cases had cytology and core-needle biopsy and 53 cases had concurrent flow cytometric analysis. FNA (cytology and flow cytometry) and core-needle biopsy were evaluated independently. FNA was diagnostic of diffuse large B-cell lymphoma (DLBL) in 25% (13/53) of cases and small B-cell NHL in 15% (8/53) of cases, whereas core-needle biopsy was diagnostic of DLBL in 37% (27/74) of cases and small B-cell NHL in 8% (6/74) of cases. Subclassification of small B-cell NHL was reached in 3/6 cases by core-needle biopsy. Insufficient cases were observed in both FNA (47%; 25/53) and core-needle biopsy (28%; 21/74) groups. With the combination of FNA and core-needle biopsy, diagnostic cases of DLBL increased to 43% (32/74) and insufficient samples were reduced to 16% (12/74). There was no clear advantage in the diagnosis and classification of small B-cell NHL by adding core-needle biopsy to FNA (14%; 10/74). We conclude that core-needle biopsy is a useful adjunct to FNA in the diagnosis of DLBL and shall be encouraged. In small B-cell NHL, core-needle biopsy does not add to the diagnostic ability of FNA. Cases insufficient for diagnosis may be seen in both core-needle biopsy and FNA. A combined approach reduces the number of insufficient cases and is recommended in routine FNA practice.     

Comparative analysis of immunoglobulin polymerase chain reaction and flow cytometry in fine needle aspiration biopsy differential diagnosis of non‐Hodgkin B‐cell lymphoid malignancies

Single primer pair polymerase chain reaction (PCR) assays for the detection of clonal immunoglobulin heavy chain (IgH) gene rearrangements and immunophenotyping by flow cytometry have been proved as useful techniques in the diagnosis of lymphoid disorders in fine needle aspirates. However, a comparative analysis of both ancillary techniques in the same samples has not been previously performed. To compare the sensitivity of flow cytometry and PCR techniques, we made a wide prospective study of 77 fine needle aspiration biopsy (FNAB) samples from lymph nodes and extranodal lymphoid infiltrates. The adjunctive values of a single primer pair PCR amplification of IgH genes and of the immunophenotyping by flow cytometry were evaluated comparing their results with the final clinicopathological diagnosis of each patient supported by histological features and clinical follow up. Among the 24 B‐cell non‐Hodgkin lymphomas, monoclonal IgH bands were detected in 22 cases by PCR, and 21 cases were correctly considered B‐cell lymphoma by flow cytometry. A monoclonal IgH band was also detected in 1 of the 53 reactive lymphoid disorders. When both ancillary techniques were combined with morphological findings, 23 of the 24 B‐cell lymphomas were correctly diagnosed but one reactive lymphoid disorder was also considered a B‐cell lymphoma. We demonstrate a similar level of detection of B‐cell lymphomas by single round PCR and flow cytometry techniques, and a strong adjunctive value when combined with morphological findings to diagnose correctly lymphoproliferative disorders by FNAB. However, we must be cautious with PCR results since false‐positive cases can occur. Diagn. Cytopathol. 2009. © 2009 Wiley‐Liss, Inc.     

Fine-needle aspiration cytology of lymphoproliferative lesions involving the major salivary glands

Fine-needle aspiration biopsy (FNA) is an accurate and cost-effective procedure for evaluating salivary gland lesions. Lymphoproliferative lesions may manifest as salivary gland enlargement. We report our experience with 43 cases of reactive and neoplastic lymphoproliferative lesions of the salivary glands evaluated by FNA, including 23 cases of reactive lymphoid hyperplasia and 20 neoplastic lymphoproliferative processes. The latter included 2 multiple myelomas and 18 non-Hodgkin lymphomas (small lymphocytic lymphoma/chronic lymphocytic leukemia, 1; small cleaved cell lymphoma, 1; lympho-plasmacytoid lymphoma, 1; mucosa-associated lymphoid tissue lymphoma, 2; mixed cell lymphoma, 4; lymphoblastic lymphoma, 1; and large cell lymphoma, 8). There were no false-negative diagnoses. Aspiration smears from 3 patients with reactive lymphoid hyperplasia and 4 patients with malignant lymphoma initially were interpreted as atypical lymphoid proliferations or as suggestive of malignant lymphoma. Thus, FNA had a sensitivity of 100% and a specificity of 87%. The majority of patients were treated medically without surgical intervention. Among the patients who underwent surgical resection of the salivary gland, 7 had an equivocal cytologic diagnosis and 2 had a benign cytologic diagnosis, but their parotid swelling failed to regress despite medical treatment. In most instances, FNA provides useful information for subsequent disease management and obviates surgical intervention.