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. 2012 Jan;14(1):43-52.
doi: 10.1093/neuonc/nor172. Epub 2011 Oct 12.

Role of flow cytometry immunophenotyping in the diagnosis of leptomeningeal carcinomatosis

Dolores Subirá  1 , Cristina Serrano, Susana Castañón, Raquel Gonzalo, Julia Illán, Javier Pardo, María Martínez-García, Esther Millastre, Francisco Aparisi, Miguel Navarro, Manuel Dómine, Ignacio Gil-Bazo, Pedro Pérez Segura, Miguel Gil, Jordi Bruna
Affiliations

Role of flow cytometry immunophenotyping in the diagnosis of leptomeningeal carcinomatosis

Dolores Subirá et al. Neuro Oncol. 2012 Jan.

Abstract

Purpose: To explore the contribution of flow cytometry immunophenotyping (FCI) in detecting leptomeningeal disease in patients with solid tumors.

Experimental design: Cerebrospinal fluid (CSF) samples from 78 patients who received a diagnosis of epithelial-cell solid tumors and had clinical data suggestive of leptomeningeal carcinomatosis (LC) were studied. A novel FCI protocol was used to identify cells expressing the epithelial cell antigen EpCAM and their DNA content. Accompanying inflammatory cells were also described. FCI results (positive or negative for malignancy) were compared with those from CSF cytology and with the diagnosis established by the clinicians: patients with LC (n = 49), without LC (n = 26), and undetermined (n = 3).

Results: FCI described a wide range of EpCAM-positive cells with a hyperdiploid DNA content in the CSF of patients with LC. Compared with cytology, FCI showed higher sensitivity (75.5 vs 65.3) and negative predictive value (67.6 vs 60.5), and similar specificity (96.1 vs 100) and positive predictive value (97.4 vs 100). Concordance between cytology and FCI was high (Kp = 0.83), although misdiagnosis of LC did not show differences between evaluating the CSF with 1 or 2 techniques (P = .06). Receiver-operator characteristic curve analyses showed that lymphocytes and monocytes had a different distribution between patients with and without LC.

Conclusion: FCI seems to be a promising new tool for improving the diagnostic examination of patients with suspicion of LC. Detection of epithelial cells with a higher DNA content is highly specific of LC, but evaluation of the nonepithelial cell compartment of the CSF might also be useful for supporting this diagnosis.

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Figures

Fig. 1.
Fig. 1.
Fig. 2.
Fig. 2.
Dot-plots showing an EpCAM positive CSF sample (A–F) and a negative sample for LC (G, H). Consecutive steps for data analysis: first step (A): identification of the CSF cell compartment based on the selection of DRAQ5 positive events. Second step (B): single-cell gate based on the selection of FSC-H vs FSC-A. Third step (C): identification of epithelial cells (ep) based on expression of EpCAM. Forth step (D, E): confirmation of a higher DNA content on epithelial cells with respect to lymphocytes DNA content. Fifth step (F): identification of inflammatory cells: lymphocytes (ly) and monocytes (mo). Epithelial cells (ep) have a heterogeneous size (FSC, forward scatter), and granularity (SSC, side scatter) as compared to lymphocytes and monocytes. No positive cells for the mAb Ber-EP4 and EpCAM are found in G and H. Positive staining for CD14 identifies monocytes. FITC: fluorescein isothiocyanate; PE: phycoerythrin.
Fig. 3.
Fig. 3.
FCI data from patients in the TP FCI group. Epithelial cells are painted in dark black, and inflammatory cells in grey. In all cases, cytology was informed as no data of malignancy. Volume of CSF sample received, CSF cell-count and percentage of malignant cells detected by FCI in every patient are specified. FSC: forward scatter; SSC: side scatter; FITC: fluorescein isothiocyanate; PE: phycoerythrin.
Fig. 4.
Fig. 4.
ROC analyses for the different percentage of lymphocytes (A) and monocytes (B) described in all cases included in the study.
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References

    1. Chamberlain MC, Glantz M, Groves MD, Wilson WH. Diagnostic tools for neoplastic meningitis: detecting disease, identifying patient risk, and determining benefit of treatment. Semin Oncol. 2009;36(4, Suppl 2):S35–S45. doi:10.1053/j.seminoncol.2009.05.005. - DOI - PubMed
    1. Gleissner B, Chamberlain MC. Neoplastic meningitis. Lancet Neurol. 2006;5(5):443–452. doi:10.1016/S1474-4422(06)70443-4. - DOI - PubMed
    1. Strik H, Prömmel P. Neoplastic meningitis. Diagnosis and individualized therapy. Expert Rev Anticancer Ther. 2010;10(7):1137–1148. doi:10.1586/era.10.86. - DOI - PubMed
    1. Bruna J, González L, Miró J, Velasco R, Gil M, Tortosa A. Leptomeningeal carcinomatosis: prognostic implications of clinical and cerebrospinal fluid features. Cancer. 2009;115(2):381–389. doi:10.1002/cncr.24041. - DOI - PubMed
    1. Shapiro RS, Johanson CE, Boogera W. Treatment modalities for leptomeningeal metastases. Sem Oncol. 2009;36(4 Suppl 2):S46–54.6. doi:10.1053/j.seminoncol.2009.05.006. - DOI - PubMed

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