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Deepened flow cytometry (FCM) analysis of bronchoalveolar lavage fluid (BALF) cells can disclose clonal B and abnormal T lymphocytes in case of lung involvement in non Hodgkin lymphoma (NHL). The possible role of routine FCM BALF analysis in diagnosing NHL involving the lungs is largely undetermined. To evaluate whether differences exist, within FCM screening of lymphocyte subsets, between BALFs from lung NHL and BALFs from reactive diseases. We compared alveolar leukocyte and lymphocyte data obtained using flow cytometry in 17 lung NHL cases with the median corresponding data detected in 208 controls, matched with cases for computed tomography findings. Absolute leukocyte counts did not differ significantly between cases and controls. As calculated within leukocytes, percentages of total, B, T, CD4+ and CD8+ T lymphocytes, respectively, were significantly higher in B cell NHL cases than in their controls (P = .003, .023, .009, .004, and .020, respectively). Similarly, percentages of total, and CD8+ T lymphocytes, respectively, were significantly higher in T cell NHL cases than in their controls (P = .046 and .027, respectively). Huger BALF lymphocytosis occurs in pulmonary NHLs than in other lung diseases. Possible lymphocyte cutoffs, that could indicate lung NHL and the subsequent need of a second-step BALF staining shortly following the initial screening, should be prospectively attempted.

An independent clinical assessment was compared with flow cytometry (FCM) and cytomorphology results obtained on 227 cerebrospinal fluids investigated for hematologic malignancy, in a retrospective longitudinal study with a median observation time of 11 months. A combined method assessment (CMA), defining "positive" a sample if at least one method gave "positive" results, was also tested. Eleven out of 55 screening samples and 53 out of 166 follow-up samples resulted positive at clinical evaluation. FCM and CM were concordant with positive clinical assessment in 68.5% and 51.5% of cases, respectively. According to CMA, 10.5% of samples (resulting false negative by either FCM or cytomorphology) were rescued as true positive. FCM retained significantly higher accuracy than cytomorphology (p = 0.0065) and 100% sensitivity when at least 220 leukocytes were acquired. CMA accuracy was higher than FCM accuracy and significantly higher than cytomorphology accuracy in the analysis of all samples (p < 0.0001), samples from mature B/T cell neoplasms (p = 0.0021), and samples drawn after intrathecal treatment (p = 0.0001). When acquiring ≥220 leukocytes, FCM accuracy was poor, and combining cytomorphology added statistically significant diagnostic advantage (p = 0.0043). Although FCM is the best diagnostic tool for evaluating CSF, morphology seems helpful especially when clinically positive follow-up samples are nearly acellular.

Flow cytometry and cytomorphology results on 92 body cavity fluids [61 effusions and 31 bronchoalveolar lavage fluids (BALF)] from hematologic malignancy were compared with retrospective clinical outcome. We observed double true positive/negative results in 67 cases (73%), and double false negative results in 2 cases (2%). Immunophenotyping accounted for true positive/negative results in 22 out of 23 mismatched cases (25%), and retained significantly higher accuracy than that of cytomorphology especially in effusions and differentiated lymphoma. In BALF analysis, immunophenotyping and cytomorphology sensitivity was 75% and 0%, respectively. Flow cytometry retains the highest accuracy in detecting neoplastic cells in body cavity fluids.

We studied circulating (C)CD34 + cells by flow cytometry in 96 patients with myelodisplastic syndromes (MDS) at diagnosis, and in a subset of 35 cases during follow-up. CCD34 + counts were stratified within both International Prognostic Scoring System (IPSS) and World Health Organization (WHO) categories. Counts >10/μl were associated with poorer leukemia-free survival, a prognostic value for evolution independent from that of WHO, and a higher progression probability within intermediate-risk IPSS and WHO classes. When serial measurements were performed, counts >10/μl more frequently correlated to evolution. Separating newly diagnosed patients on the basis of 10/μl cut-off of circulating CD34 + cells retains prognostic utility, especially in intermediate-risk MDS.

Introduction: Flow cytometry is well known to detect malignant cells in peripheral blood and bone marrow of patients with hematologic malignancies. However, its role in evaluating non-hematic body fluid contamination by tumor cells is largely unexplored. Patients and Methods: Data detected by flow cytometry in non-hematic body fluid samples drawn between 2002 and 2007 from patients with hematologic neoplasms were retrospectively compared with morphological findings obtained from cytospin slides. Immunophenotyping was carried out by using disease-specific multicolor panels of quadruple monoclonal antibodies, conjugated with the fluorochromes FITC, PE, PerCP, and APC, respectively. Acquisition of information on 1x104 to 1x105 stained cells depending on the whole sample cellularity was assessed on a dual-laser FACSCalibur flow cytometer using the CellQUEST software (Becton Dickinson, San José, CA, USA). Results: Fourty-five samples (bronchoalveolar fluid, n=5; ascites, n=2; hydrocele, n=2; pleural effusion, n=8; and cerebrospinal fluid, n=28) from 32 patients were available for comparison. Diagnoses were as follows: chronic myelomonocitic leukemia (n=1), acute promyelocytic leukaemia (n=2), acute myelomonocytic leukaemia (n=1), B-chronic lymphocitic leukemia (n=4), follicular lymphoma (n=1), acute lymphoblastic leukaemia (n=6), lymphoblastic lymphoma (n=1), high grade non-Hodgkin’s lymphoma (NHL), Burkitt-like (n=3), diffuse large B-cell NHL (n=6), peripheral T-cell NHL (n=3), and NHL, unspecified (n=4). Flow cytometry detected neoplastic cells in 24 cases. Of these cases, only 17 were positive also by morphology. In 7 cases, in which tumor cells were detected by flow cytometry but not by morphology, clinical data confirmed the presence of the disease. Flow cytometry did not show neoplastic cells in 21 cases. Of these cases, only 18 were negative also by morphology. In the remaining 3, the suggestion of diffuse large B-cell NHL contamination by morphology was not confirmed by flow cytometry, demonstrating T-reactive lymphocytes that were clearly negative for disease-specific markers. Conclusions: Our data suggest that flow cytometry is a useful tool complementary to morphology for the screening of non-hematic body fluid contamination in patients with hematologic neoplasms.