Detection of minimal residual disease in mantle cell lymphoma-establishment of novel eight-color flow cytometry approach.

BACKGROUND: Minimal residual disease (MRD) detection is an essential tool for therapy response assessment in a considerable number of hematooncologic disorders, including mantle cell lymphoma (MCL). Flow cytometry (FCM) ranks among the most effective approaches, which allows rapid sample processing and compete successfully with highly sensitive molecular methods like polymerase chain reaction. Because FCM is ordinarily applied to detect MRD in B-lineage diseases like chronic lymphocytic leukemia, a similar method could be used in MCL. We decided to test our novel eight-color FCM approach in MCL MRD detection. METHODS: Using an eight-color FCM protocol designed by us, the expression of 24 selected surface antigens in a cohort of 30 patients with newly diagnosed leukemic MCL and 20 normal controls were compared to establish markers that can reliably distinguish normal B-lymphocytes from the MCL population. The sensitivity of the designed protocol was tested using serial dilution studies. RESULTS: Although MCL presents high immunophenotypic variability, the combination of CD20/23/5/19/200/62L/3/45 seems to be very favorable in flow cytometric MRD measurement. CONCLUSIONS: Our eight-color FCM protocol could easily detect MRD in MCL patients, and reaches a sensitivity of up to 2 × 10(-4) .

Detection of Minimal Residual Disease in Mantle Cell Lymphoma. Establishment of Novel 8-Color Flow Cytometry Approach.

Backround: Minimal residual disease (MRD) detection is an essential tool for therapy response assessment in a considerable number of hematooncologic disorders including mantle cell lymphoma (MCL). Flow cytometry (FCM) ranks among the most effective approaches which allow rapid sample processing and compete successfully with highly sensitive molecular methods like polymerase chain reaction (PCR). Since FCM is ordinarily applied to detect MRD in B-lineage diseases like chronic lymphocytic leukemia (CLL), a similar method could be used in MCL. We decided to test our novel 8-color FCM approach in MCL MRD detection. Methods: Employing an 8-color FCM protocol designed by us, the expression of 24 selected surface antigens in a cohort of 30 patients with newly diagnosed leukemic MCL and 20 normal controls were compared to one another to establish markers that can reliably distinguish normal B lymphocytes from the MCL population. The sensitivity of the designed protocol was tested using serial dilution studies. Results: Although MCL presents high immunophenotypic variability the combination of CD20/23/5/19/200/62L/3/45 appears to be very favorable in flow cytometric MRD measurement. Conclusions: Our 8-color FCM protocol could easily detect MRD in MCL patients and reaches a sensitivity of up to 2×10-4 . This article is protected by copyright. All rights reserved.