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May 2000, Volume 14, Number 5, Pages 922-930
Table of contents    Previous  Abstract  Next   Full text  PDF
Original Manuscript
A novel natural killer cell line (KHYG-1) from a patient with aggressive natural killer cell leukemia carrying a p53 point mutation
M Yagita1, C L Huang2, H Umehara3, Y Matsuo4, R Tabata1, M Miyake2, Y Konaka1 and K Takatsuki1

1Department of Clinical Immunology and Hematology, Tazuke-Kofukai Medical Research Institute, Kitano Hospital, Osaka, Japan

2Department of Thoracic Surgery, Tazuke-Kofukai Medical Research Institute, Kitano Hospital, Osaka, Japan

3Department of Internal Medicine, Osaka Dental University, Osaka, Japan

4Fujisaki Cell Center, Hayashibara Biochemical Labs, Inc., Okayama, Japan

Correspondence to: M Yagita, Department of Clinical Immunology and Hematology, Tazuke-Kofukai Medical Research Institute, Kitano Hospital, 13-3 Kamiyama-cho, Kita-ku, Osaka 530-8480, Japan; Fax: 81-6-6361-0588

Abstract

We present the establishment of a natural killer (NK) leukemia cell line, designated KHYG-1, from the blood of a patient with aggressive NK leukemia, which both possessed the same p53 point mutation. The immunophenotype of the primary leukemia cells was CD2+, surface CD3-, cytoplasmic CD3epsilon+, CD7+, CD8alphaalpha+, CD16+, CD56+, CD57+ and HLA-DR+. A new cell line (KHYG-1) was established by culturing peripheral leukemia cells with 100 units of recombinant interleukin (IL)-2. The KHYG-1 cells showed LGL morphology with a large nucleus, coarse chromatin, conspicuous nucleoli, and abundant basophilic cytoplasm with many azurophilic granules. The immunophenotype of KHYG-1 cells was CD1-, CD2+, surface CD3-, cytoplasmic CD3epsilon+, CD7+, CD8alphaalpha+, CD16-, CD25-, CD33+, CD34-, CD56+, CD57-, CD122+, CD132+, and TdT-. Southern blot analysis of these cells revealed a normal germline configuration for the beta, delta, and bold gamma chains of the T cell receptor and the immunoglobulin heavy-chain genes. Moreover, the KHYG-1 cells displayed NK cell activity and IL-2-dependent proliferation in vitro, suggesting that they are of NK cell origin. Epstein-Barr virus (EBV) DNA was not detected in KHYG-1 cells by Southern blot analysis with a terminal repeat probe from an EBV genome. A point mutation in exon 7 of the p53 gene was detected in the KHYG-1 cells by PCR/SSCP analysis, and direct sequencing revealed the conversion of C to T at nucleotide 877 in codon 248. The primary leukemia cells also carried the same point mutation. Although the precise role of the p53 point mutation in leukemogenesis remains to be clarified, the establishment of an NK leukemia cell line with a p53 point mutation could be valuable in the study of leukemogenesis. Leukemia (2000) 14, 922-930.

Keywords

cell line; NK cell leukemia; p53; point mutation

Received 11 October 1999; accepted 20 January 2000
May 2000, Volume 14, Number 5, Pages 922-930
Table of contents    Previous  Abstract  Next   Full text  PDF