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Sensitivity and Resistance to Therapy

In vitro IL-12 treatment of peripheral blood mononuclear cells from patients with leukemia or myelodysplastic syndromes: increase in cytotoxicity and reduction in WT1 gene expression

Leukemia volume 14, pages 1634–1641 (2000)Cite this article

Abstract

Interleukin-12 (IL-12) has potent antitumor activities. We examined whether IL-12 enhanced the cytotoxicity of peripheral blood mononuclear cells (PBMNC) and decreased leukemia cells in 30 patients with leukemia or myelodysplastic syndromes (MDS): 12 acute myeloid leukemia (AML) (five in complete remission (CR) and seven in non-CR); six chronic myeloid leukemia (CML); and 12 MDS (three refractory anemia (RA), eight RA with excess of blasts and one chronic myelomonocytic leukemia). PBMNC from patients and five healthy volunteers were cultured at 5 × 105/ml parallel with or without 100 units/ml of IL-12 for 3 days. Cytotoxicity of PBMNC against K562 cells was assessed by flow cytometry. To quantify the amount of leukemia cells, WT1 mRNA was measured by competitive reverse transcription polymerase chain reaction (RT-PCR), since WT1 mRNA is considered as a marker of minimal residual disease (MRD) in leukemia or MDS. The cytotoxicity of non-IL-12-treated PBMNC of 30 patients was 13.4 ± 9.3% at the effector to target (E:T) ratio of 20:1, and significantly lower than that of normal subjects (25.7 ± 8.4%). The cytotoxicity increased to 30.6 ± 17.9% in the IL-12-treated PBMNC. WT1 mRNA in PBMNC of five healthy volunteers was less than 103 copies/μg of total RNA. Following the 3-day IL-12 treatment, mean WT1 mRNA of PBMNC was reduced from 104.8 to 104.2copies/μg of total RNA in six CML patients, from 105.4 to 104.8copies/μg in 12 MDS patients and from 105.0 to 104.2 copies/μg in five AML patients in CR, but not reduced in five of seven AML in non-CR. These results showed that IL-12 significantly enhanced PBMNC cytotoxicity and decreased the quantity of leukemia cells in PBMNC of most patients with MDS, CML and AML in CR. IL-12 might be of considerable benefit in the elimination of MRD in patients with hematological malignancies. Leukemia (2000) 14, 1634–1641.

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Acknowledgements

We thank Dr M Kobayashi (Center Molecular Biology and Cytogenetics, SRL, Japan) for his kind technical advice about competitive RT-PCR. We also thank Dr Quang-Kim Tran for his helpful proofreading of the manuscript.

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  1. Department of Medicine III, Hamamatsu University School of Medicine, 3600, Handa-cho, Hamamatsu, 431–3192, Japan

    L Pan, K Ohnishi, WJ Zhang, H Yoshida, L Maksumova, F Muratkhodjaev, K Shigeno, S Nakamura, JM Luo, HL Hao, S Fujisawa, K Naito, K Shinjo, A Takeshita & R Ohno

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Pan, L., Ohnishi, K., Zhang, W. et al. In vitro IL-12 treatment of peripheral blood mononuclear cells from patients with leukemia or myelodysplastic syndromes: increase in cytotoxicity and reduction in WT1 gene expression. Leukemia 14, 1634–1641 (2000). https://doi.org/10.1038/sj.leu.2401872

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