[CANCER RESEARCH 54, 4430-4435, August 15, 1994] Abnormal Cytokine Expression in Sézary and Adult T-Cell Leukemia Cells Correlates with the Functional Diversity between These T-Cell Malignancies1 Craig L. Tendier,2 Jack D. Burton, Jonathan Jatte, David Danielpour, Michael Charley, J. Philip McCoy, Mark R. Pittelkow, and Thomas A. Waldmann Division of Pediatrie Oncology, Mount Sinai School of Medicine, New York, New York 10029-6574 /C. L. T./; Metabolism Branch ¡J.D. B., T. A. W.] and Laboratory of Chemoprevention [D. D.], National Cancer Institute, NIH, Bethesda, Maryland 20892; Division of Allergy and Pulmonary Medicine, Hahnemann University, Philadelphia, PA 19102 [J. J.¡; Department of Dermatology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15260 fM. C, J. P. M.]; and Department of Dermatology, Mayo Clinic, Rochester, Minnesota 55905 [M. R. W.] ABSTRACT TheSézarysyndrome (SzS) and adult T-cell leukemia (ATL) are malig nant proliferations of mature T-lymphocytes that possess distinct func tions. Sézarycells function as helper cells, whereas ATL cells are usually suppressor effectors. Although phenotypically similar (CD4+/CD7-/ CDS—), these functional differences between the T-cell lymphoprolifera- tive disorders suggest different patterns of cytokine expression. We wished to delineate the cytokine mechanisms potentially underlying the diverse functional characteristics of SzS and ATL. Therefore, we analyzed the expression of interleukins (IL) 2, 4, and 5, y-interferon, and transforming growth factor /!, in the highly purified leukemic T-cells from 5 SzS and 5 ATL patients. Decreased inKNA and protein levels of IL-2, y-interferon, and 11-5 were detected in mitogen-stimulated ATL and SzS cells when compared to similarly cultured normal CD4+ cells. In contrast, IL-4 production was markedly up-regulated in the leukemic cells of 4/5 SzS patients as compared to ATL and normal controls. Finally, fresh ATL cells secreted higher levels of transforming growth factor ß,into the culture medium than the malignant T-cells from SzS patients. Collectively these results suggest that, similar to the murine CD4-expressing T-cell subsets Thl and I h2, different cytokine profiles exist in a human popu lation of CD4+ T-cells. Moreover, the distinct patterns of IL-4 and transforming growth factor ß,expression by SzS and ATL cells, respec tively, are most consistent with the functional differences (i.e., helper versus suppressor activity) between these T-cell malignancies and thus may play important roles in the pathogenesis of the paraneoplastic fea tures associated with these two leukemias. INTRODUCTION The malignant expansion of mature CD4+ T-cells is represented clinically by two distinct human leukemias known as the SzS3 and ATL. SzS is a cutaneous T-cell lymphoma associated with diffuse skin involvement known as erythroderma, generalized lymphadenop- athy, and circulating malignant cells in the blood (1, 2). ATL occurs in patients infected with the retrovirus HTLV-I and is also character ized by infiltration of the skin with malignant T-cells (3). However, it tends to have a more aggressive course than SzS and is often com plicated by hepatosplenomegaly, hypercalcemia, and an extremely high incidence of opportunistic infections (4, 5). The circulating leukemic cells in SzS and ATL patients share the Received 3/31/94; accepted 6/13/94. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 Supported in part by a Young Investigator Award from the Mount Sinai Children's Health Research Center. NICHD Grant, 5P30 HD28822 (C. L. T.). 2 To whom requests for reprints should be addressed, at PédiatrieHematology/Oncol- ogy, Mount Sinai Medical Center, Box 1208, One Gustave Levy Place, New York, NY 10029. 3 The abbreviations used are: SzS, Sézary syndrome; ATL, adult T-cell leukemia; IL-2, interleukin 2; IFN-7, -y-interferon; TGF-/3,, transforming growth factor ß,;PHA, phyto- hemagglutinin; PMA, phorbol 12-myristate 13-acetate; PCR, polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay; HTLV-I, human T-cell leukemia virus type I; PBMC, peripheral blood mononuclear cell; cDNA, complementary DNA. identical mature T-cell phenotype, CD4+, but lack CD7 and CDS antigen expression (6, 7). Although phenotypically similar, biological differences between these T-lymphoproliferative disorders suggest an altered pattern of cytokine expression. For example, in immunoglobulin biosynthesis assays the neoplastic T-cells from SzS patients function as helper cells when cocultured with stimulated normal B-cells, whereas in the same assay ATL cells effectively suppress normal immunoglobulin production (8). Moreover, peripheral blood lymphocytes from ATL pa tients have impaired proliferative responses to T-cell mitogens (5). This in vitro immunosuppressor activity of ATL cells is paralleled clinically by the uniform finding of cutaneous anergy, failure to make antibody to infused murine monoclonal antibodies, and increased incidence of op portunistic infections in ATL patients (5, 9-10). In contrast, lack of anergy, elevated levels of IgE, and the production of antibodies following antigenic stimulation have all been demonstrated in patients with SzS (1, 2). These functional and clinical immunological differences between the CD4+ leukemic T-cells from SzS and ATL patients suggest that, similar to the murine T-helper cell populations designated Th 1 and Th2, distinc tive cytokine profiles exist among human CD4-expressing T-cell subsets as well. Recently, mitogen-activated peripheral blood mononuclear cells isolated from SzS patients were shown to produce significantly higher protein levels of IL-4 and lower levels of IL-2 and IFN-y than did similarly cultured cells from normal controls (11). However, this study did not provide direct evidence that it is the malignant Sézary cell which is responsible for the abnormal immune response since enriched CD4+ leukemic populations from SzS patients were not analyzed. Moreover, there has been no investigation comparing the cytokine profile in SzS with respective levels of cytokine expression in ATL to better define their relevance to the pathogenesis of the immunological abnormalities associated with these T-cell malignan cies. Therefore, we have examined the expression of IL-2, IFN-y, IL-4, IL-5, and TGF-ß, in highly purified populations of leukemic T-cells isolated from SzS and ATL patients to determine which cytokines are potential mediators of the diverse immune abnormalities observed in these T-cell lymphoproliferative disorders and to gain insight into the regulatory network of cells that control the human immune response. MATERIALS AND METHODS Patients. Five patients with SzS and five with ATL were studied. Healthy blood bank donors, all males with a mean age of 46 years (range, 25-70), served as normal controls. Informed consent was obtained from all patient and volunteer blood donors. The SzS group consisted of four males and one female from the United States with a mean age of 64 years (range, 50-71). This patient population was characterized by the presence of erythroderma, lymphadenopathy, and a circulating pool of abnormal lymphocytes with cere- briform nuclei and a predominant CD4+/CD7-/CD8- T-cell phenotype. Except for one SzS patient with a total WBC of 7,100/mm3, leukocytosis was a consistent finding in this group ranging from 18,100 to 45,800 cells/mm3. All 4430 Research. on November 26, 2021. © 1994 American Association for Cancer cancerres.aacrjournals.org Downloaded from