Differential Signaling and Hierarchical Response Thresholds Induced by an Immunodominant Peptide of Myelin Basic Protein and an Altered Peptide Ligand in Human T Cells Paul D. Crowe, Stefen A. Boehme, Tim Wong, Amitabh Gaur, John Sidney, Alessandro Sette, and Paul J. Conlon ABSTRACT: Changes in peptide antigen concentration or structure can have a profound effect on T cell respon- siveness by inducing selected T cell effector functions. In this study, we have compared the biological responses of an MBP83-99-specific human T h 0 T cell clone (TCC) stimulated with increasing concentrations of native pep- tide or an altered peptide ligand (APL). Our results show that the hierarchy of response thresholds for proliferation and cytokine secretion is similar for native peptide and APL. However, because a much higher concentration of the APL is required to evoke the same degree of response, the cytokine profile is shifted towards a T h 2-like response relative to the same concentration of native peptide. In addition, we observed qualitative differences in TCR sig- nal transduction triggered by native peptide and a weak agonist APL even at concentrations that elicit similar biological responses. Thus, the relationship between TCR signaling and biological responses may be more complex than previously recognized. Human Immunology 59, 679 – 689 (1998). © American Society for Histocompat- ibility and Immunogenetics, 1998. Published by Elsevier Science Inc. ABBREVIATIONS MBP myelin basic protein APL altered peptide ligand EBV Epstein-Barr virus TCC T cell clone INTRODUCTION Recognition of MHC-peptide ligand complexes by T cell antigen receptors initiates a wide range of effector func- tions including clonal expansion and production of cy- tokines which determine the nature and magnitude of the ensuing immune response [1]. Functionally distinct subsets of murine T h cells have been described based upon the cytokines they secrete [2]. T h 1 cells are char- acterized by their ability to produce IFN-and TNF that enhance cellular immunity, whereas T h 2 cells se- crete IL-4, IL-5 and IL-10, promoting humoral immu- nity. For human T lymphocytes, the demarcation be- tween functional T h subtypes is not as well defined and some cells, designated T h 0, are characterized by their ability to produce both T h 1 and T h 2 cytokines. The functional subtype of a given T lymphocyte has impor- tant implications both in terms of immune responsive- ness to pathogens and autoimmunity. In the case of autoimmunity, previous work has shown that T h 1 cells play a major role in the pathogenesis of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), the murine model of the disease. Other studies have implicated T h 2 cytokines in the natural recovery from disease [3–5]. Thus, the balance of cytokines se- From Neurocrine Biosciences, Inc., 10555 Science Center Drive (P.D.C., S.A.B., T.W., A.G., P.J.C.) and Epimmune, Inc., 6555 Nancy Ridge Drive (J.S., A.S.), San Diego, CA 92121, USA. Address reprint requests to: Dr. Paul D. Crowe, Neurocrine Biosciences, Inc., 10555 Science Center Drive, San Diego, CA 92121, USA; Tel: (619) 658-7669; Fax: (619) 658-7601; E-Mail: pcrowe@neurocrine.com. Received July 6, 1998; accepted July 28, 1998. Human Immunology 59, 679 – 689 (1998) 0198-8859/98/$19.00 © American Society for Histocompatibility and Immunogenetics, 1998 Published by Elsevier Science Inc. PII S0198-8859(98)00077-9