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Updated: Apr 5, 2026
An Efficient and Simple Method to Establish NK and T Cell Lines from Patients with Chronic Active Epstein-Barr Virus Infection
Published on: March 30, 2018
Establishment and Characterization of a CD20-Positive NK/T-Cell Lymphoma Cell Line
Background:
CD20 positive NK/T-cell lymphoma is extremely rare and difficult for clinical treatment. Due to the lack of an established cell model for this disease, less is known about its biological characterization and potential therapeutic options.
Methods:
A cell line of NK/T-cell lymphoma, which was enriched by magnetic sorting with proper cell surface markers (CD56) from peripheral blood mononuclear cells (PBMCs) drawn from a 21-year-old male patient with nasal angiocentric NK/T-cell lymphoma, was designated as ZQNK-29. Immunophenotypic analysis of ZQNK-29 was performed by flow cytometric and immunohistochemical analysis. Comparative genomic hybridization (CGH) analysis was used for cytogenetic analysis of ZQNK-29. Potential rearrangements of the immunoglobulin gene and Epstein-Barr virus (EBV) infection were examined by PCR and RT-PCR, respectively.
Results:
ZQNK-29 cells express the phenotypic T-cell marker (CD3), T cell activation markers (HLA-DR), markers for both NK and cytotoxic T lymphocytes (TIA-1), and B-lineage marker CD20; however, expression of CD56 was not detected in expanded ZQNK-29 cells although this NK cell surface marker was used as one of selective cell surface markers for the initial isolation of NK/T cells. RT-PCR analysis showed that the pattern of gene expressions for infected EBV was latency type III, with the expressions of LMP1, EBNA-1, and EBNA-2; no rearrangements were found in the heavy-chain of the immunoglobulin gene or in the y chain of the T cell receptors (TCRs) gene. CGH analysis demonstrated that ZQNK-29 possessed an abnormal karyotype, 46XY, 1p (dist)+, 4p (dist)+, 4q (mid)-, 5q (mid)-, 9q (dist)+, 16p (dist)+, 16q (dist)+, 17p+, 17q (dist)+, 19q (dist)+, 20p+, 20q+, 21q+, and 22q+. Of these, 1p (dist)+, which has been confirmed to be mitochondrial DNA amplification, is believed to be mainly caused by EBV infection.
Conclusions:
ZQNK-29 is a well characterized premature human NK/T-cell lymphoma cell line with expression of the B-cell marker CD20 and will provide a useful pre-clinic model for characterization and potential therapeutic studies of the aggressive NK/T-cell lymphoma.
Insights
A new cell line, ZQNK-29, was established from a patient with rare CD20-positive NK/T-cell lymphoma. This model aids research into the disease
Area of Science:
- Hematology
- Oncology
- Immunology
Background:
- CD20-positive NK/T-cell lymphoma is a rare and challenging malignancy.
- Lack of established cell models hinders understanding and treatment development.
Purpose of the Study:
- To establish and characterize a novel cell line for CD20-positive NK/T-cell lymphoma.
- To provide a pre-clinical model for investigating biological characteristics and therapeutic strategies.
Main Methods:
- Established ZQNK-29 cell line from patient peripheral blood mononuclear cells (PBMCs).
- Performed immunophenotypic analysis (flow cytometry, immunohistochemistry).
- Conducted cytogenetic analysis (CGH), PCR for gene rearrangements, and RT-PCR for Epstein-Barr virus (EBV) infection.
Main Results:
- ZQNK-29 cells express CD3, HLA-DR, TIA-1, and CD20, but not CD56.
- EBV infection was present with latency type III gene expression (LMP1, EBNA-1, EBNA-2).
- Abnormal karyotype identified, with 1p amplification potentially linked to EBV.
Conclusions:
- ZQNK-29 is a well-characterized human NK/T-cell lymphoma cell line expressing CD20.
- This cell line serves as a valuable pre-clinical model for aggressive NK/T-cell lymphoma research.
