Isolation and characterization of a CD34+ sub-clone in B-cell lymphoma
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Ayad M. Al-Katib1, Abdul Shukkur Ebrahim1, Mustapha Kandouz2, Feras Zaiem2, Ali Raufi1, Salah Ebrahim2, Anwar Mohamed2, Nada Emara1 and Ali M. Gabali2
1 Lymphoma Research Laboratory, Department of Internal Medicine, Wayne State University School of Medicine, Detroit, MI 48201, USA
2 Department of Pathology, Wayne State University School of Medicine, Detroit, MI 48201, USA
|Ayad M. Al-Katib,||email:||firstname.lastname@example.org|
Keywords: CD34; B-cell non-Hodgkin’s lymphoma; minimal residual disease; lymphoma stem cell
Received: July 26, 2019 Accepted: December 02, 2019 Published: January 14, 2020
Non-Hodgkin’s lymphoma (NHL) is the most common hematological malignancy in the US. Many types remain incurable despite response to initial therapy and achievement of complete remission (CR). Advanced laboratory techniques like multicolor flow cytometry (FCM) and polymerase chain reaction (PCR) have demonstrated persistence of rare malignant cell population post therapy. However, the functional and biological characteristics of this population have not been elucidated.
Established B-lymphoma cell lines (B-NHL) and patient-derived samples (PDS) were analyzed using 8-color FCM. CD34+ sub-population was enriched using in vitro exposure to 2-chlorodeoxyadenosine (2-CdA) and by CD34 magnetic beads. Genetic analysis of cell fractions was done by karyotyping and array comparative genomic hybridization (aCGH). Sensitivity to chemotherapy was assayed by short-term in vitro exposure to chemotherapy. Clonogenicity was determined by soft agar colony formation assay, and proliferation was determined using DNA staining with propidium iodide and FCM.
FCM demonstrated the presence of a minute sub-clone of monotypic B-cells that express CD34 in B-NHL cell lines (3 of 3) and in PDS (8 of 8). This sub-population enriched up to 50 fold in vitro by exposure to 2-CdA and up to 80% purity by CD34 magnetic bead column isolation. Except for CD34 expression, this population expressed identical phenotype and genotype to parent cells, but was more proliferative, Hoechst 33342-positive, clonogenic, and resistant to chemotherapy compared with the CD34- population.
The isolated CD34+ monotypic B-cells may contribute to resistance of certain NHL to treatment and should be targeted by potential new drugs for NHL.
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