title: Brain stereotactic biopsy flow cytometry for central nervous system lymphoma characterization: advantages and pitfalls.
abstract: Brain stereotactic biopsy (SB) followed by conventional histopathology and immunohistochemistry (IHC) is the gold standard approach for primary central nervous system lymphoma (PCNSL) diagnosis. Flow cytometry (FCM) characterization of fine-needle aspiration cytology and core needle biopsies are increasingly utilized to diagnose lymphomas however, no biological data have been published on FCM characterization of fresh single cell suspension from PCNSL SB. The aim of this study was to establish the feasibility and utility of FCM for the diagnosis and characterization of brain lymphomas from a tissue samples obtained by a single SB disaggregation.
  
  
  
  Twenty-nine patients with a magnetic resonance suggestive for PCNSL entered the study. A median of 6 SB were performed for each patient. A cell suspension generated from manual tissue disaggregation of a single, unfixed, brain SB, was characterized by FCM. The FCM versus standard approach was prospectively compared.
  
  
  
  FCM and IHC showed an high degree of agreement (89 %) in brain lymphoma identification. By FCM, 16 out of 18 PCNSL were identified within 2 h from biopsy. All were of B cell type, with a heterogeneous CD20 mean fluorescence intensity (MFI), CD10 positive in 3 cases (19 %) with surface Ig light chain restriction documented in 11 cases (69 %). No false positive lymphomas cases were observed. Up to 38 % of the brain leukocyte population consisted of CD8 reactive T cells, in contrast with the CD4 positive lymphocytes of the peripheral blood samples (P < 0.001). By histopathology, 18 B-PCNSL, only one CD10 positive (5 %), 1 primitive neuroectodermal tumor (PNET) and 10 gliomas were diagnosed. A median of 6 days was required for IHC diagnosis.
  
  
  
  Complementary to histopathology FCM can contribute to a better characterization of PCNSL, although necrosis and previous steroid treatment can represent a pitfall of this approach. A single brain SB is a valid source for accurate FCM characterization of both lymphoma and reactive lymphocyte population, routinely applicable for antigen intensity quantification and consistently documenting an active mechanism of reactive CD8 T-lymphocytes migration in brain lymphomas. Moreover, FCM confirmed to be more sensitive than IHC for the identification of selected markers.
authors: Clinical Pathology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Iole
authors: I
authors: Cordone
authors: Clinical Pathology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Serena
authors: S
authors: Masi
authors: Pathology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Mariantonia
authors: M
authors: Carosi
authors: Radiology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Antonello
authors: A
authors: Vidiri
authors: Hematology and Stem Cell Transplant, Regina Elena National Cancer Institute, Rome, Italy.
authors: Francesco
authors: F
authors: Marchesi
authors: Pathology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Mirella
authors: M
authors: Marino
authors: Neurosurgery, Regina Elena National Cancer Institute, Rome, Italy.
authors: Stefano
authors: S
authors: Telera
authors: Clinical Pathology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Alessia
authors: A
authors: Pasquale
authors: Hematology and Stem Cell Transplant, Regina Elena National Cancer Institute, Rome, Italy.
authors: Andrea
authors: A
authors: Mengarelli
authors: Clinical Pathology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Laura
authors: L
authors: Conti
authors: Pathology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Edoardo
authors: E
authors: Pescarmona
authors: Neuroncology, Regina Elena National Cancer Institute, Rome, Italy.
authors: Andrea
authors: A
authors: Pace
authors: Neurosurgery, Regina Elena National Cancer Institute, Rome, Italy. carmine.carapella@ifo.gov.it.
authors: Carmine M
authors: CM
authors: Carapella
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044963:Biopsy, Fine-Needle; D062005:Biopsy, Large-Core Needle; D001932:Brain Neoplasms / Q000175:diagnosis*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005910:Glioma / Q000175:diagnosis*; D006801:Humans; D008223:Lymphoma / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D017599:Neuroectodermal Tumors / Q000175:diagnosis*; D011446:Prospective Studies; D012680:Sensitivity and Specificity; D055815:Young Adult
keywords: Brain stereotactic biopsy; Flow cytometry; PCNSL; Tumor side population