Lymph node FNA cytology: Diagnostic performance and clinical implications of proposed diagnostic categories.

BACKGROUND: Despite widespread clinical use, lymph node fine-needle aspiration cytology (LN-FNAC) lacks universal acceptance for definitively diagnosing lymphomas. This is likely due to reports of lower diagnostic performance, inconsistent terminology use in cytopathology diagnostic reports, and only limited data on the clinical implications of LN-FNAC diagnoses. Recently, a uniform LN-FNAC cytopathological diagnostic reporting system was proposed (the Sydney System). This study evaluated LN-FNAC diagnostic performance and risks of malignancy associated with the proposed diagnostic categories. METHODS: LN-FNAC specimens obtained in 2018-2019, with and without concurrent core biopsy, to evaluate for suspected lymphoma were analyzed (n = 349). LN-FNAC diagnoses were compared with final diagnoses obtained via subsequent tissue biopsy and/or clinical assessment. RESULTS: The mean patient age was 57.6 years, and 41% were female. LN-FNAC was the initial diagnostic test in 223 (63.9%), and it was used to evaluate for recurrence in 126 (36.1%). LN-FNAC diagnosed 202 hematological malignancies (57.9%), 23 nonhematological malignancies (6.6%), and 124 reactive processes (35.5%). Subsequent tissue biopsy was performed in 42 (12%). The risks of malignancy per diagnostic category were as follows: inadequate, 58.3%; benign, 6.4%; atypical, 69.2%; suspicious, 96.7%; and malignant, 99.3%. LN-FNAC demonstrated up to 96.3% sensitivity, 91.91% specificity, and 87.35% accuracy. Optimal specimen quality and the use of intradepartmental consultation reduced diagnostic error rates in FNA cases without concurrent core biopsy (P = .029 and P = .0002 respectively). CONCLUSIONS: LN-FNAC is accurate and reliable for the diagnosis of lymphoma. Inadequate LN-FNAC samples should be resampled due to a significant associated risk of lymphoma. The diagnostic performance of LN-FNAC may be improved with good specimen quality and reviews by multiple pathologists. Understanding the risks of malignancy associated with LN-FNAC diagnostic categories will help to guide optimal patient management.

Diagnostic utility of cerebrospinal fluid flow cytometry in patients with and without prior hematologic malignancy.

Flow cytometry (FCM) is an adjunct study to routine analysis of cerebrospinal fluid (CSF) to investigate for involvement by a hematologic malignancy. However, in our experience, FCM only infrequently detects abnormalities in CSF. To help optimize resources without forfeiting clinically important data, we sought to determine evidence-based indications and criteria for performing FCM on CSF. FCM results of 316 consecutive CSF specimens were retrospectively reviewed and correlated with clinical history, total nucleated cell (TNC) counts, and results of concurrent cytologic review. Of 255 samples adequate for analysis, 54% were from patients with a prior history of hematologic malignancy, of which 12% (17 cases) were abnormal by FCM. Corresponding TNC counts among samples with abnormal FCM ranged from 0-1050 cells/µL, and only 44% showed abnormal morphology on concurrent cytology. Of the remaining 46% of samples from patients with no known history of hematologic malignancy who had CSF sampling for neurological indications, only one (1%) was abnormal by FCM. This specimen had an elevated TNC count (39 cells/µL) but lacked clearly abnormal findings on concurrent cytology. These results support the use of CSF FCM only in patients with a history of hematologic malignancy or, in the absence of such a history, in samples showing pleocytosis. If these criteria were applied to the current cohort using a TNC count cut-off of >5 cells/µL, 23% of samples would have been deferred from testing, resulting in decreased cost, improved efficiency, and reduction in the need for unnecessary testing without a negative impact on clinical care.

Flow cytometry for hematopoietic cells.

Within the last 25 years, flow cytometry and fluorescence-activated cell sorting have emerged as both routine diagnostic tools in clinical medicine and as advanced analytic tools critical in performing scientific research. This chapter aims at summarizing the use of flow cytometry in benign and malignant hematology and the monitoring of inherited and acquired immunodeficiency states. Numerous figures are provided from our laboratories at Massachusetts General Hospital that illustrate examples of these conditions. The chapter also describes novel flow cytometry-based imaging techniques, the combination of flow cytometry and mass spectrography, new software tools, and some future directions and applications of advanced instrumentation for flow cytometry.

Flow cytometry is of limited utility in the early identification of "double-hit" B-cell lymphomas.

BACKGROUND: B-cell lymphomas with concurrent translocations of MYC and BCL2 or BCL6, also known as "double-hit" lymphomas (DHL), are rare malignancies characterized by aggressive clinical behavior and poor prognosis. Previous reports suggest that decreased CD20 and/or CD19 expression by flow cytometry is relatively common in DHL and may help to identify cases requiring additional cytogenetic analysis. METHODS: We conducted a retrospective analysis of 26 cases of DHL, and compared their flow cytometric characteristics to cases of Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL). Cases were analyzed by four-color flow cytometry, and bivariate dot-plots were reviewed for light scatter characteristics, CD19, CD20, CD45, and surface light chain. RESULTS: Relatively few DHL cases showed dim expression of CD19 or CD20, and statistically significant differences were found only in the frequency of dim CD19 expression between DHL and BL or DLBCL. Although concomitant dim CD19 and CD20 expression was exclusive to DHL, it was present in only a minority of cases. CONCLUSIONS: We conclude that although a subset of DHL expresses aberrant levels of CD19 and/or CD20 by flow cytometry, these findings are of limited utility in identifying cases requiring cytogenetic analysis due to their low frequency. Until more sensitive pathologic parameters can be identified and validated, the decision to perform cytogenetic analysis should rest on a combination of clinical, morphologic, and immunophenotypic features suggestive of high-grade, aggressive disease.

Relationship between ABO genotype and A antigen expression on platelets.

BACKGROUND: Although platelets (PLTs) are known to express ABH antigens, the extent of expression is different on PLTs compared with red blood cells and the relationship between PLT ABH expression and genotype has not been thoroughly investigated. STUDY DESIGN AND METHODS: We measured blood group H and A antigens on PLTs from 100 normal volunteers using fluorescent-conjugated reagents and flow cytometry. Individuals were also genotyped at the ABO locus using a commercially available genotyping system. RESULTS: Expression of A and H antigen varied widely on PLTs from different individuals. Among group A and AB persons, H antigen expression was significantly greater than A antigen (p < 0.0001). The ratio of H-to-A antigen expression varied more than 100-fold in a predictable fashion according to genotype with values lowest in A1/A1 < A1/O < A2/A2 < A2/O. H and A antigen expression was unaffected by secretor status. The proportion of PLTs with high A expression also varied directly according to genotype. CONCLUSIONS: Blood group A and H antigen expression on PLTs varies in a predictable fashion according to genotype. Flow cytometry and genotyping identify individuals who strongly express A antigens, a finding that may be relevant to clinical PLT transfusion across ABO barriers.

Soluble CD163, a novel marker of activated macrophages, is elevated and associated with noncalcified coronary plaque in HIV-infected patients.

BACKGROUND: Pro-inflammatory monocytes/macrophages may contribute to increased atherosclerosis in human immunodeficiency virus (HIV)-infected patients. We investigate--to our knowledge, for the first time--sCD163 and other markers of monocyte activation in relationship to atherosclerotic plaque in HIV-infected patients. METHODS: One hundred two HIV-infected and 41 HIV-seronegative men with equivalent cardiovascular risk factors and without history of coronary artery disease were prospectively recruited and underwent computed tomography coronary angiography. RESULTS: sCD163 levels and presence of plaque were significantly higher among antiretroviral-treated subjects with undetectable HIV RNA levels, compared with seronegative controls (1172 ± 646 vs. 883 ± 561 ng/mL [P = .02] for sCD163 and 61% vs. 39% [P = .03] for presence of plaque). After adjusting for age, race, lipids, blood pressure, glucose, smoking, sCD14, and HIV infection, sCD163 remained independently associated with noncalcified plaque (P = .008). Among HIV-infected patients, sCD163 was associated with coronary segments with noncalcified plaque (r = 0.21; P = .04), but not with calcium score. In contrast, markers of generalized inflammation, including C-reactive protein level, and D-dimer were not associated with sCD163 or plaque among HIV-infected patients. CONCLUSIONS: sCD163, a monocyte/macrophage activation marker, is increased in association with noncalcified coronary plaque in men with chronic HIV infection and low or undetectable viremia. These data suggest a potentially important role of chronic monocyte/macrophage activation in the development of noncalcified vulnerable plaque. CLINICAL TRIAL REGISTRATION: NCT00455793.