Abstract
Approximately 65% (11/17) of cancer patients participating in an ongoing Phase I clinical trial with recombinant interleukin-2 developed nonneutralizing serum IgG anti-interleukin-2 antibodies within 1 month of initiating therapy. These antibodies could be detected using any of several standard techniques including immunoblots and enzyme-linked immunosorbent assays. Western blot analysis and retention experiments with protein A-Sepharose indicate that the antibodies are specific for interleukin-2. The interleukin-2 mutein utilized in this clinical trial (des-ala-ser125 r-IL-2) differs from the major species of the human T cell-derived lymphokine in that it lacks the N-terminal alanine of the native molecule, is not glycosylated, and possesses a serine-cysteine substitution at position 125. Another recombinant interleukin-2, identical to the mutein except that it retains the cysteine at position 125 (des-ala-cys125 r-IL-2), strongly competes with the mutein in competitive enzyme-linked immunosorbent assays, suggesting that the amino acid substitution is not responsible for the recognition of the molecule by serum antibodies. Conversely, nonrecombinant T cell-derived interleukin-2 fails to compete in these assays and is not retained by protein A-Sepharose columns when mixed with high-titer antiserum. These results suggest that the anti-interleukin-2 serum antibodies generated in the course of treatment do not react with the nonrecombinant lymphokine but recognize epitopes peculiar to recombinant forms which are not dependent on the amino acid substitution at position 125. The failure of the antibodies to neutralize the biological activity of recombinant interleukin-2 (IL-2) in lymphocyte proliferation assays and to bind to the native lymphokine suggests that they may not affect IL-2-dependent cellular immune functionsin vivo.
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Morgan D, Ruscetti F, Gallo R: Selectivein vitro growth of T-lymphocytes from normal human bone marrows. Science 193:1007–1008, 1976
Ruscetti F, Morgan D, Gallo R: Functional and morphologic characterization of human T-cells continuously grownin vitro. J Immunol 119:131–137, 1977
Henney C, Kuribayashi K, Kern D, Gillis S: Interleukin-2 augments natural killer activity. Nature 291:353–357, 1981
Dempsey R, Dinarello C, Mier J, Rosenwasser L, Allegretta M, Brown T, Parkinson D: The differential effects of human leukocytic pyrogen/lymphocyte activating factor, T-cell growth factor and interferon on human natural killer activity. J Immunol 129:2504–2510, 1982
Howard M, Matis L, Malek T, Shevach E, Kell W, Cohen D, Nakanishi K, Paul W: Interleukin-2 induces antigen-reactive T-cell lines to secrete BCGF-1. J Exp Med 158:2024–2034, 1983
Dennert G, Yogeeswaran G, Yamagata S: Cloned cell lines with natural killer activity. J Exp Med 153:545–563, 1981
Grimm E, Mazumder A, Zhang Z, Rosenberg S: Lymphokine-activated killer cell phenomenon. I. Lysis of natural killer-resistant fresh solid tumor cells by interleukin-2-activated autologous human peripheral blood lymphocytes. J Exp Med 155:1823–1832, 1982
Grimm E, Robb R, Roth J, Neckers L, Lachman L, Wilson D, Rosenberg S: Lymphokine-activated killer cell phenomenon. III. Evidence that IL-2 is sufficient for direct activation of peripheral blood lymphocytes into lymphokine-activated killer cells. J Exp Med 158:1356–1361, 1983
Grimm E, Ramsey K, Mazumder A, Wilson D, Djeu J, Rosenberg S: Lymphokine-activated killer cell phenomenon. II. Precursor phenotype is serologically distinct from peripheral T-lymphocytes, memory cytotoxic thymus-derived lymphocytes, and natural killer cells. J Exp Med 157:884–897, 1983
Mazumder A, Rosenberg S: Successful immunotherapy of natural killer-resistant established pulmonary melanoma metastases by the intravenous adoptive transfer of syngeneic lymphocytes activatedin vitro by interleukin-2. J Exp Med 159:495–510, 1984
Hefeneider S, Conlon P, Henney C, Gillis S:In vivo Interleukin-2 administration augments the generation of alloreactive cytolytic T-lymphocytes and resident natural killer cells. J Immunol 130:222–228, 1983
Rosenberg S, Mulé J, Spiess P, Reichert C, Schwarz S: Regression of established pulmonary metastases and subcutaneous tumor mediated by the systemic administration of high dose recombinant interleukin-2. J Exp Med 161:1169–1182, 1985
Taniguchi T, Matsui H, Fugita T, Takaoka C, Kashima N, Yoshimoto R, Hamuro J: Structure and expression of a cloned cDNA for human interleukin-2. Nature 302:305–308, 1983
Wang A, Lu S, Mark D: Site-specific mutagenesis of the human interleukin-2 gene: Structure-function analysis of the cysteine residues. Science 224:1431–1434, 1984
Gutterman J, Fine S, Quesada J, Horning S, Levine J, Alexanian R, Bernhardt C, Kramer K, Spiegel H, Colburn W, Trown P, Merigan T, Dziewanoska Z: Recombinant leukocyte A interferon: Pharmacokinetics, single-dose tolerance, and biological effects in cancer patients. Ann Intern Med 96:549–558, 1982
Vallbracht A, Treuner J, Flehmig B, Joester K, Niethammer D: Interferon-neutralizing antibodies in a patient treated with human fibroblast interferon. Nature 289:496–499, 1981
Rosenberg S, Grimm E, McGrogan M, Doyle M, Kawasaki E, Koths K, Mark D: Biological activity of recombinant human interleukin-2 produced inEscherichia coli. Science 223:1412–1415, 1984
Glass W, Briggs R, Hnilica L: Identification of tissue-specificnuclear antigens transferred to nitrocellulose from polyacrylamide gels. Science 211:70–72, 1981
Gillis S, Ferm M, Ou W, Smith K: T-cell growth factor: Parameters of production and a quantitative microassay for activity. J Immunol 120:2027–2032, 1978
Smith K, Favata M, Oroszian S: Production and characterization of monoclonal antibodies to human interleukin-2: Strategy and tactics. J Immunol 131:1808–1815, 1983
Lotze M, Frana L, Sharrow S, Robb R, Rosenberg S:In vivo administration of purified human interleukin 2. I. Half-life and immunologic effects of the Jurkat cell line-derived interleukin 2. J Immunol 134:157–162, 1985
Lotze M, Matory Y, Fettinghausen S, Rayner A, Sharrow S, Seipp C, Custer M, Rosenberg S:In vivo administration of purified human interleukin 2. II. Half-life, immunologic effects, and expansion of peripheral lymphoid cellsin vivo with recombinant IL-2. J Immunol 135:2865–2872, 1985
Parem S, Check I, Henriksen D, Vilcek J: Antibodies to interferon in a patient with systemic lupus erythematosus. J Immunol 129:1–7, 1982
Wolf R, Andreoni J: Soluble inhibitory factor (SIF) in normal human serum. Cell Immunol 67:299–311, 1982
Kaufman D, Carnaud C, Stach J, Bach J: The suppressive effect of a supernate from concanavalin A-activated human lymphocytes: Effects of concanavalin A-activated lymphocytes and their supernates on cytotoxic and mixed lymphocyte reactions. Cell Immunol 47:153–162, 1979
Granelli-Piperno A, Andrus L, Reich E: Antibodies to interleukin-2. Effects on immune responsesin vitro andin vivo. J Exp Med 160:738–751, 1984
Atkins M, Gould J, Allegretta M, Li J, Dempsey R, Rudders R, Parkinson D, Reichlin S, Mier J: Phase-I evaluation of recombinant interluekin-2 in patients with advanced malignant disease. J Clin Oncol (in press)
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Allegretta, M., Atkins, M.B., Dempsey, R.A. et al. The development of anti-interleukin-2 antibodies in patients treated with recombinant human interleukin-2 (IL-2). J Clin Immunol 6, 481–490 (1986). https://doi.org/10.1007/BF00915254
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DOI: https://doi.org/10.1007/BF00915254