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Quantification and cytokine production of circulating lymphoid and myeloid cells in acute myelogenous leukaemia

Leukemia volume 17, pages 716–730 (2003)Cite this article

Abstract

A simple assay was developed to assess the potential of patients with acute myelogenous leukaemia (AML) to respond to immunotherapy. Lymphocytes, monocytes and leukaemic blasts with their corresponding intracellular cytokine profiles were evaluated by four-colour flow cytometry. In 50 μl samples of whole blood, surface labelling for CD45, CD8 and CD3 was used for cell identification prior to intracellular staining for interleukin (IL)-4, IL-10, IL-12 and interferon (IFN)-γ. Absolute numbers of CD8+ and CD8− (putative CD4+) T-cells, NK cells (CD8+/CD3−) and monocytes were determined by reference to a fixed number of added fluorescent beads. The absolute numbers of CD8− and CD8+ T-cells in the blood of patients with AML were similar to those of normal controls. More of the lymphocytes in the blood of leukaemic patients spontaneously produced cytokines compared with those of controls. Furthermore, primary AML blasts secreted predominantly IFN-γ. After recovery from chemotherapy, lymphocyte counts tended to be lower than in normals and reduction of NK cells reached significance after the second chemotherapy (P=0.01). A prominent CD8lo/CD3lo-int lymphocyte subset appeared after recovery in some patients. This laboratory application of the study of cell subsets and intracellular cytokines in patients undergoing treatment may be helpful in monitoring immunological responses in AML.

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Acknowledgements

We thank the patients who kindly donated the samples and the medical staff of the Haematology Department and the Lister Unit at Northwick Park Hospital for collecting them. We also thank Dr John Farrant and Margaret North for assistance with flow cytometry acquisition and analysis.

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Authors and Affiliations

  1. Antigen Presentation Research Group, Faculty of Medicine, Imperial College London, Northwick Park Campus, London, UK

    N Panoskaltsis & S C Knight

  2. Department of Haematology, Northwick Park Hospital, Harrow, London, UK

    N Panoskaltsis & C D L Reid

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  1. N Panoskaltsis
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  2. C D L Reid
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  3. S C Knight
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NP is supported by the Leukaemia Research Fund and the Northwick Park Leukaemia Research Trust Fund; SCK is supported by the Medical Research Council, UK

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Panoskaltsis, N., Reid, C. & Knight, S. Quantification and cytokine production of circulating lymphoid and myeloid cells in acute myelogenous leukaemia. Leukemia 17, 716–730 (2003). https://doi.org/10.1038/sj.leu.2402835

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