Cytokine Responsiveness of Mitogen-Activated T Cells Derived from Acute Leukemia Patients with Chemotherapy-Induced Leukopenia

Abstract

The aim of the study was to characterize effects of exogenous cytokines on T lymphocytes derived from acute leukemia patients with chemotherapy-induced leukopenia. We investigated the cytokine responsiveness of long-term expanded CD4+ and CD8+ T cell clones and the effects of exogenous cytokines on anti-CD3-stimulated polyclonal T cell responses. After mitogenic activation in the presence of acute myelogenous leukemia (AML) blasts, most CD4+ and CD8+ clones proliferated in response to interleukin-2 (IL-2). Although a majority of the IL-2-responsive clones could also proliferate in the presence of exogenous IL-4, IL-7, IL-9, IL-10, IL-12, and IL-15, only IL-15 responses were equal to or exceeded the corresponding IL-2 responses. Exogenous cytokines were also added during T cell activation with phytohemagglutinin (PHA) + accessory leukemia cells derived from different AML patients, and all the cytokines then had divergent effects that depended on both differences between clones and differences between AML patients. However, for most of these T cell clone/AML blast combinations, IL-2 and IL-15 caused enhanced T cell proliferation. IL-2 and IL-15 also enhanced anti-CD3-stimulated polyclonal responses of nonexpanded T cells derived from cytopenic patients, whereas other cytokines had only minor effects. Our results demonstrate that cytokine-responsive T cells remain in the circulation during chemotherapy-induced cytopenia, and combination therapy including intensive chemotherapy and T cell-targeting cytokine therapy should, therefore, be possible in AML.

In vitro culture of human osteosarcoma cell lines: a comparison of functional characteristics for cell lines cultured in medium without and with fetal calf serum

“…The methods for preparation and characterisation of the clones have been described in detail previously (Bruserud et al 1998;Bruserud and Ulvestad 2000a;Bruserud and Ulvestad 2000b). All the clones expressed the membrane molecule phenotype CD2 + CD4 + CD8 À T cell receptor (TCR) ab + cd À .…”

“…The release of a wide range of CCL and CXCL chemokines was also investigated for Cal72 and SJSA-1 cells (for details, see below). Alternatively, 3 H-thymidine was added after 48 h (37 kBq/well added in 20 ll saline; TRA 310, Amersham International, Amersham, UK) and nuclear radioactivity assayed 18 h later (Bruserud and Ulvestad 2000a;Bruserud and Ulvestad 2000b).…”

“…Based on our previous studies (Bruserud and Ulvestad 2000a;Bruserud and Ulvestad 2000b) 2·10 4 CTC were incubated in 170 ll medium in each well of U-bottomed microtiter plates (Costar 3796) with 5·10 4 sarcoma cells/ well and phytohemagglutinin (PHA) 1 lg/ml (HA 16, Wellcome, Dartford, UK). Supernatants were harvested after 2 days, and the levels of Interferon-c (IFN-c; minimal detectable level 5 pg/ml) and GM-CSF were determined by ELISA analysis (Quantikine ELISA kits, R&D Systems).…”

Functional Evaluation of Proliferative T Cell Responses in Patients with Severe T Lymphopenia: Characterization of Optimal Culture Conditions and Standardized Activation Signals for a Simple Whole Blood Assay

“…The quantitative T cell defects in such patients can be characterized by membrane molecule analysis of the remaining T cell population, whereas the methods for evaluation of qualitative defects and especially the qualitative component of mixed defects are less well characterized and standardized. Very few previous studies have therefore investigated T cell functions in patients with severe T lymphopenia, e.g., cancer patients with severe cytopenia after intensive chemotherapy or autologous stem cell transplantation (2,3).…”

The Immunological Dilemma: Cellular Innate and Adaptive Immune Response Versus Human Acute Myeloid Leukemia

“…There have also been some reports regarding qualitative/functional evaluation of T cells, these include the examination of cytokine release and cytokine responsiveness of in vitro expanded clonogenic T cells derived from AML patients with therapy-induced cytopenia [129][130][131][132][133]. These clonogenic cells constitute a minor population of circulating T cells including CD4 + and CD8 + TCR + cells [130], and the cells secrete a wide range of immunoregulatory cytokines [131].…”