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*
Howard Hughes Medical Institute and Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305;
Nuffield Department of Clinical Medicine, University of Oxford, Headington, United Kingdom; and
Third Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan
We describe the generation of three mAbs that recognize the complex of the class II MHC molecule IEk bound to a peptide derived from the carboxyl terminus of moth cytochrome c (residues 95–103). Reactivities of these mAbs are sensitive to single alterations in the sequence of both helices of the MHC molecule and to the bound peptide. The epitopes of these reagents are distinct but overlap substantially. One of these mAbs specifically blocks lymphokine release by T cells responsive to this complex but not others. We have used another to examine how the number of complexes on an APC is related to its ability to stimulate T cells. We find that 200–400 complexes per cell are necessary and sufficient to induce a degree of stimulation, whereas maximum stimulation is achieved only if more than 5000 complexes are present. The analysis indicates that T cell activation is a stochastic process.
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