Characterization of a New Chronic Lymphocytic Leukemia Cell Line for Mechanistic In Vitro and In Vivo Studies Relevant to Disease

Abstract

Studies of chronic lymphocytic leukemia (CLL) have yielded substantial progress, however a lack of immortalized cell lines representative of the primary disease has hampered a full understanding of disease pathogenesis and development of new treatments. Here we describe a novel CLL cell line (OSU-CLL) generated by EBV transformation, which displays a similar cytogenetic and immunophenotype observed in the patient’s CLL (CD5 positive with trisomy 12 and 19). A companion cell line was also generated from the same patient (OSU-NB). This cell line lacked typical CLL characteristics, and is likely derived from the patient’s normal B cells. In vitro migration assays demonstrated that OSU-CLL exhibits migratory properties similar to primary CLL cells whereas OSU-NB has significantly reduced ability to migrate spontaneously or towards chemokine. Microarray analysis demonstrated distinct gene expression patterns in the two cell lines, including genes on chromosomes 12 and 19, which is consistent with the cytogenetic profile in this cell line. Finally, OSU-CLL was readily transplantable into NOG mice, producing uniform engraftment by three weeks with leukemic cells detectable in the peripheral blood spleen and bone marrow. These studies describe a new CLL cell line that extends currently available models to study gene function in this disease.

Ibrutinib enhances chimeric antigen receptor T-cell engraftment and efficacy in leukemia

“…This cell line has the immunophenotype and migratory properties similar to the primary CLL tumor from which it was derived. 19 In this system, based on the engraftment of 5 to 10 million B-CLL cells, neither single-agent ibrutinib nor CTL019 affected overall survival. In contrast, the combination of these agents resulted in both markedly enhanced CTL019 engraftment in the peripheral blood ( Figure 6G) and significant prolongation of survival ( Figure 6H).…”

“…19 Wild-type parental K562 cells (purchased from American Type Culture Collection), K562 cells engineered to express CD19 (K562-CD19) along with PD-L1 (K562-CD19-PDL1), and Nalm-6 cells (American Type Culture Collection) were used as targets where indicated in the relevant experiments. Primary donor T cells were transduced with the anti-CD19 CAR containing a 4-1BB intracellular signaling domain (CTL019) and expanded as previously reported.…”

IPI-145 antagonizes intrinsic and extrinsic survival signals in chronic lymphocytic leukemia cells

“…10 The retroviral construct pRetroX-Tight-Puro was used to stably transfect XLA cells with BTK. A mutation was made using QuikChange site directed mutagenesis (Stratagene) in the kinase domain at cysteine 481 to serine (see the primer sequence in supplemental Methods on the Blood Web site).…”

BRAFV600E accelerates disease progression and enhances immune suppression in a mouse model of B-cell leukemia

“…29 This cell line displays immunophenotype and cytogenetics similar to the patient's CLL (trisomy 12 and 19). OSUCLL cells with doxycycline-inducible expression of normal or mutated BRAF were previously described.…”