A recombinant antibody with the antigen-specific, major histocompatibility complex-restricted specificity of T cells.

Abstract

Specific recognition of peptide/major histocompatibility complex (MHC) molecule complexes by the T-cell receptor is a key reaction in the specific immune response. Antibodies against peptide/MHC complexes would therefore be valuable tools in studying MHC function and T-cell recognition and might lead to novel approaches in immunotherapy. However, it has proven difficult to generate antibodies with the specificity of T cells by conventional hybridoma techniques. Here we report that the phage display technology is a feasible alternative to generate antibodies recognizing specific, predetermined peptide/MHC complexes.T and B cells represent two fundamentally different recognition modes of the specific immune system. Through alternating selection processes T cells are educated to recognize antigenic peptides presented in association with self-molecules of the major histocompatibility complex (MHC) on the surface of antigen-presenting cells. In contrast, B cells are not educated to be self-MHC-restricted and B-cell receptors (antibodies), whether soluble or in membrane-bound form, recognize threedimensional target structures. The distinctly different education of B and T cells explains why antibodies with the MHCrestricted specificity of T cells are rare and why it has been difficult to generate such specificities by conventional B-cell hybridoma techniques. We have taken advantage of the selection power of the phage display technology which makes it possible to test tens of millions of individual clones and have devised a method to generate recombinant antibodies recognizing predetermined peptide/MHC complexes. The speed and feasibility of this method makes it realistic to produce antibodies to a variety of specific peptide/MHC complexes which may be useful in studying MHC-restricted T-cell recognition and may lead to novel approaches in diagnostics and immunotherapy.MATERIALS AND METHODS MHC Purification. The AKR mouse-derived lymphoma RDM-4 was used for Kk production as described (1). In brief, Kk molecules were immunoaffinity purified from detergent cell lysates by using the monoclonal anti-Kk antibody 11.4-1 (American Tissue Type Culture Collection). The affinity columns were washed extensively and bound MHC class I molecules were eluted with 0.05 M diethylamine, pH 11/0.15 M sodium chloride/0.1% sodium azide/0.1% sodium deoxycholate, neutralized, and concentrated by vacuum dialysis. Human 82-microglobulin was obtained from the urine of uremic patients and purified to homogeneity by gel filtration and chromatofocusing (1).The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.The influenza virus-derived nucleoprotein peptide NP50-57 (single-letter code, SDYEGRLI) and hemagglutinin peptide Ha255-262 (FESTGNLI) were synthesized manually on a RaMPS synthesizer (DuPont) using standard fluorenylmethoxycarbonyl protection strategy.Generation of Peptide/MHC Cl...

Direct Phenotypic Analysis of Human MHC Class I Antigen Presentation: Visualization, Quantitation, and In Situ Detection of Human Viral Epitopes Using Peptide-Specific, MHC-Restricted Human Recombinant Antibodies

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Levels of Specific Peptide-HLA Class I Complex Predicts Tumor Cell Susceptibility to CTL Killing

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